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Ishrat Majid Bababode Adesegun Kehinde Basharat Dar Vikas Nanda Editors
Advances in Plant Sprouts
Phytochemistry and Biofunctionalities
Advances in Plant Sprouts
Ishrat Majid Bababode Adesegun Kehinde Basharat Dar • Vikas Nanda Editors
Advances in Plant Sprouts Phytochemistry and Biofunctionalities
Editors Ishrat Majid Department of Food Technology Islamic University of Science and Technology Awantipora, JK, India Basharat Dar Food Technology Islamic University of Science and Technology, Awantipora Srinagar, Jammu and Kashmir, India
Bababode Adesegun Kehinde Food Processing Center University of Nebraska-Lincoln Lincoln, NE, USA Vikas Nanda Department of Food Engineering and Technology Sant Longwal Institute of Engineering and Technology Longowal, Punjab, India
ISBN 978-3-031-40915-8 ISBN 978-3-031-40916-5 (eBook) https://doi.org/10.1007/978-3-031-40916-5 © The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Nature Switzerland AG 2023 This work is subject to copyright. All rights are solely and exclusively licensed by the Publisher, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed. The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. The publisher, the authors, and the editors are safe to assume that the advice and information in this book are believed to be true and accurate at the date of publication. Neither the publisher nor the authors or the editors give a warranty, expressed or implied, with respect to the material contained herein or for any errors or omissions that may have been made. The publisher remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. This Springer imprint is published by the registered company Springer Nature Switzerland AG The registered company address is: Gewerbestrasse 11, 6330 Cham, Switzerland Paper in this product is recyclable.
Preface
The dietary concept of vegetable foods continuously gains progressive attention at virtually all life facets. At homes, production industries, academia, religious zones, and social gatherings, consumers around the world have increased their focus on vegetable-based food materials based on diverse rationales ranging from modernistic trends to campaigns against animal slaughter and consumption. However, the strong argument persists on whether plant foods bear nutrient loads that will suffice human vitality requirements. Accordingly, efforts have been implemented in researching viable and sustainable methodologies to improve plant-based foods in making them worthy alternatives to animal diets. The operation of sprouting has been optimally studied over the years to be effective in remarkably improving plant foods as functional agents beyond their basic dietary employments. When sprouted, plant foods have been reported to be enhanced in their potential against health disorders when consumed. This course has been attributed to the changes in their phytochemical profiles during sprouting. More precisely, it has been reported that larger macromolecules of biological importance are being degraded into smaller but more therapeutically formidable units. Foods of various classes have also been found employable for the sprouting process. Diverse examples of cereals, pseudocereals, and legumes, amongst others, can be sprouted to make them dietetically, nutritionally, and therapeutically better. The necessity for a piece of scientific literature that is comprehensive to a diverse group of audience behooves the birthing of this book. This book was concocted to serve as an information base for consultation on the operation, merits, safety concerns, and study trends of plant sprouts. Several studies regarding optimized process parameters, elicitations, phytochemistry, nutritional outcomes, and sensory effects of sprouting are herein reviewed. Awantipora, Jammu and Kashmir, India Lexington, KY, USA Srinagar, Jammu and Kashmir, India Longowal, India
Ishrat Majid Bababode Adesegun Kehinde Basharat Dar Vikas Nanda
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Contents
1 General Over View of Composition, Use in Human Nutrition, Process of Sprouting, Change in Composition During Sprouting, Parameters Affecting Nutritional Quality During Sprouting, Benefits of Sprouts, Nutritional Value and Food Safety Issues of Cereals/Pseudo Cereal Sprouts���������������������������������������������������������� 1 Jyoti Duhan, Nisha Kumari, Manjeet Singh, and Bharat Garg 2 Barley Sprouts������������������������������������������������������������������������������������������ 29 Mamta Thakur and Sudha Rana 3 Buckwheat Sprouts���������������������������������������������������������������������������������� 57 D. Sowdhanya, Jyoti Singh, Prasad Rasane, Sawinder Kaur, Jaspreet Kaur, and Mukul Kumar 4 Brown Rice Sprouts: A Leading Functional Food Product������������������ 99 Bharat Garg, Shikha Yashveer, Manjeet Singh, and Jyoti Duhan 5 Amaranth & Quinoa Sprouts ���������������������������������������������������������������� 127 Anamika Sharma, Masud Alam, Kirty Pant, and Vikas Nanda 6 Oat and Kamut Sprouts�������������������������������������������������������������������������� 153 Pooja Kesarkar, Papiha Gawande, and Yogesh Gat 7 Sprouted Legumes: Biochemical Changes, Nutritional Impacts and Food Safety Concerns�������������������������������������������������������� 173 Josephine Ampofo and Lord Abbey 8 Kidney Bean Sprouts and Lentil Sprouts���������������������������������������������� 201 K. C. Dileep, Kanchan Bhatt, Satish Kumar, Rakesh Sharma, Priyanka Rana, Monika Thakur, and Priyanka Suthar 9 Clover and Alfalfa Sprouts���������������������������������������������������������������������� 229 Bababode Adesegun Kehinde, Oluwakemi Igiehon, Adekanye Oluwabori, and Ishrat Majid
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10 Black-Eyed Peas, Chickpeas and Pea Sprouts�������������������������������������� 237 Meenakshi Trilokia, Wani Suhana Ayoub, and Preeti Choudhary 11 Mung and Adzuki Bean Sprouts������������������������������������������������������������ 275 Dilpreet Singh Brar, Amritpal Kaur, and Vikas Nanda 12 Soybean Spouts: A Healthier Alternative���������������������������������������������� 299 Parv Bansal, Neha Babbar, Vikas Kumar, Sukhpreet Kaur, and Poonam Aggarwal 13 An Overview of Brassica Sprouts ���������������������������������������������������������� 313 Ankit Kumar, Ramandeep Kaur, Satish Kumar, Dharminder Kumar, Rajat Chandel, and Vikas Kumar 14 Broccoli and Cress Sprouts �������������������������������������������������������������������� 331 Puneet Kang, Sawinder Kaur, Jyoti Singh, and Prasad Rasane 15 Cabbage and Red Cabbage Sprouts: Powerhouse of Nutrients���������� 363 Shweta Sharma, Priyanka, Bharti Shree, Preethi Ramachandran, Vikas Kumar, Ramesh Thakur, and Satish Kumar 16 Radish Sprouts and Mustard Green Sprouts���������������������������������������� 383 Ankit Kumar, Ramandeep Kaur, Satish Kumar, Ramesh Thakur, Dharminder Kumar, Rajat Chandel, and Vikas Kumar 17 General Overview of Composition, Use in Human Nutrition, Process of Sprouting, Change in Composition During Sprouting, Parameters Affecting Nutritional Quality During Sprouting, Benefits of Sprouts, Nutritional Values and Food Safety Issues of Allium Sprouts�������������������������������������������������������������������������� 403 Bindu Bazaria and Neeraj 18 Onion Sprouts������������������������������������������������������������������������������������������ 417 Bababode Adesegun Kehinde, Oluwakemi Igiehon, Adekanye Oluwabori, and Ishrat Majid 19 Garlic and Leek Sprouts ������������������������������������������������������������������������ 427 Shuchi Upadhyay, Indra Rautela, Sanjay Kumar, B. S. Rawat, Vinod Kumar, and Shradha Manish Gupta Index������������������������������������������������������������������������������������������������������������������ 439
Chapter 1
General Over View of Composition, Use in Human Nutrition, Process of Sprouting, Change in Composition During Sprouting, Parameters Affecting Nutritional Quality During Sprouting, Benefits of Sprouts, Nutritional Value and Food Safety Issues of Cereals/Pseudo Cereal Sprouts Jyoti Duhan, Nisha Kumari, Manjeet Singh, and Bharat Garg
1.1 General Overview of Composition of Cerals and Pseudocereals Consumers have been more aware of the health advantages of the food they eat over the past several years. They are shifting towards “quality over quantity”, consequently looking for the food with optimum amount of bioactive compounds and sprouts are one of them. Well, sprouting exists in the life of eastern countries from a long time, and now sprouting has raised its popularity in western countries on consumers’ demand for dietetics and exotic healthy foods. Cereals are a staple meal in practically every region of the globe and are crucial to human nutrition. The three cereal species that are most often produced are rice, maize, and wheat. The pseudocereals amaranth, quinoa, canihua, and buckwheat are among the additional cereal species that exist, in addition to millet, sorghum, oat, and barley. Most people rely heavily on cereals and pseudocereals for their nutritional needs. Proteins, carbohydrates, fats, vitamins, and minerals are found in J. Duhan (*) · N. Kumari Department of Biochemistry, College of Basic Sciences & Humanities, CCS Haryana Agricultural University, Hisar, Haryana, India M. Singh Oilseeds Section, Department of Genetics & Plant Breeding, College of Agriculture, CCS Haryana Agricultural University, Hisar, Haryana, India B. Garg Department of Molecular Biology, Biotechnology & Bioinformatics, College of Basic Sciences & Humanities, CCS Haryana Agricultural University, Hisar, Haryana, India © The Author(s), under exclusive license to Springer Nature Switzerland AG 2023 I. Majid et al. (eds.), Advances in Plant Sprouts, https://doi.org/10.1007/978-3-031-40916-5_1
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abundance in these foods. Contrarily, antinutritive elements such as phytic acid, tannins, polyphenolics, and enzyme inhibitors reduce the nutritive value of grains by complexing proteins, sugars, and minerals. Cereal and pseudocereal grains contain these antinutritive elements naturally. The most popular types of cereals and pseudocereals products available in industrialised countries include bread, breakfast cereals, and cereal bars. The eating of fermented grains as drinks, cakes, or porridge is prevalent in developing nations (Kockova et al., 2013). All grains are made of an endosperm, an embryo, and a bran layer. Due to the uneven distribution of the bran and embryo throughout the grain, the milling procedure can have a substantial impact on the final grain grate composition (Slavin et al., 2000). Refined grains are lower in micronutrients than their whole grain equivalents because micronutrients are confined to the superficial layer of the grain. When compared to whole grains, processed grains have lower levels of polyphenols, phytic acid, tannin, certain enzyme inhibitors, vitamins, and minerals. Whole grains also include more dietary fibre and other fiber-associated nutrients (Oghbaei & Prakash, 2016; Slavin et al., 2000). Since sprouted grains have a greater potential for containing essential nutrients, flavours, and textural qualities than non-germinated grains, they are becoming more and more popular. Sprouted grains include mung beans and black beans, as well as oats, millets, barley, rice, wheat, corn, rye and sorghum. Germinated oat, barley, brown rice, wheat and sorghum are among the most popular sprouted grains. When comparing pseudocereal grains to true cereals, one can detect a striking similarity in their starch content and physical appearance (Alvarez-Jubete et al., 2010a). Pseudocereals are desirable crops for the future because of their diverse genetic makeup, which allows them to be adapted to a variety of climates, from tropical to temperate (Ruiz et al., 2014; Joshi et al., 2018, 2019). Carbohydrate content in quinoa, canihua, amaranth, and buckwheat ranges from 48.5 to 77.0 percent dm (dry mass), 60–70 percent dm, and 63.1–70.0 percent dm, and 63.1–82.2 percent, respectively. The starch level in amaranth is 55–65%; in quinoa, 52.2–72%; in canihua, 48–51%; and in buckwheat, 54.5–78% (Repo-Carrasco-Valencia et al., 2010; Martınez-Villaluenga et al., 2020). As compared to actual cereals, the quantities are a little smaller. Pseudocereals, such as amaranth, quinoa, and canihua, have greater simple carbohydrate contents than other cereals, ranging from 3% to 5% (Pereira et al., 2019), whereas buckwheat has lower simple carbohydrate contents. Grains vary widely in their protein content, with some having as little as 8% and others as high as 16% of dry mass. Pseudocereals’ high protein content is one of its nutritional advantages. The protein is concentrated in the endosperm. Exceptional quality is attributed to this protein by sulfur-rich amino acids and a well-balanced mixture of necessary amino acids. Pseudocereal protein fractions tend to look more like legume proteins than cereal proteins in terms of distribution (Janssen et al., 2017). With the exception of buckwheat, pseudocereals’ total lipid content is often greater (Canihua: 5.0–8.0%; amaranth: 5.6–10.9%; buckwheat: 0.75–7.7%; quinoa: 4.0–7.6%). The majority of the lipids in all three grains are composed of unsaturated fatty acids, with linoleic and oleic acids being the most prevalent (Amaranth: 61.0–87.3%, buckwheat: 80.1–80.9%, quinoa: 71.0–84.5%) (De Bock et al., 2021).
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Minerals are abundant in whole pseudocereal grains since the bran contains most of the mineral components. Potassium, phosphorus, and magnesium are the most common pseudocereal minerals. Celiacs, who are more susceptible to osteopenia and osteoporosis, may benefit from amaranth’s high calcium content in particular (Martínez-Villaluenga et al., 2020). The vitamin composition of pseudocereals is also intriguing, as they contain a high amount of B vitamins. Quinoa seeds are rich in vitamin B6 and folate, making them ideal for both children and adults (USDA, 2005). Furthermore, quinoa seeds’ riboflavin content meets 40% of the daily requirements for adults and 80% of the daily requirements for children (USDA, 2005). Non-nutrient plant components, such as bioactive chemicals, have just lately been discovered to have health advantages. Phytosterols, phytoestrogens, polysaccharides, phytoecdysteroids, bioactive proteins and betalains are some of the bioactive components found in pseudocereal grains.
1.2 Use in Human Nutrition To consume the sprouts, the seed must be germinated in water or another medium, then the sprouts must be harvested before the real leaves have formed and eaten whole with the seed. The practise of sprouting seeds is not new, especially in Eastern countries where seedlings have a long culinary history. As interest in dietetics and unusual healthy foods increased in the 1980s, sprouted seed eating became increasingly popular in Western countries. In recent years, sprouted seed interest has centered on minimal processing and additive-free processing. A wide range of premium foods, like salads and garnishes, can take advantage of their distinguishing characteristics such as their peculiar colour, rich flavour, and considerable richness in bioactive components (Treadwell et al., 2010). Sprouting is an easy and affordable approach that uses very little room in the greenhouse, doesn’t need sophisticated equipment, and produces high yields (Delian et al., 2015; Kyriacou et al., 2016). Beyond malting, which is an alcoholic beverage-specific kind of germination, grain seedlings can be eaten as sprouts or undergo additional processing, such as drying or roasting (Hübner & Arendt, 2013). Bread made from sprouting grains and pseudocereals could be on the rise (Falcinelli et al., 2018). It’s possible that sprouted cereals’ physical qualities and baking performance could be adversely affected by excessive levels of enzyme activity during uncontrolled germination (Marti et al., 2018). They can also be used in the preparation of noodles and pasta, laddue, unleavened bread and porridge (Shingare & Thorat, 2013). It’s feasible that functional beverages, such as those made from a mixture of sprouted grains and flour, could be the future. The resistant starch, oligosaccharides, and water-soluble fibres present in cereals have been recommended as a way to help probiotic compositions fill their void. Fresh juices, pills and liquid concentrates of wheatgrass are the most common methods of ingesting it. The addition of cereal sprouts to animal feed, as has been suggested for non-grain species, could provide additional perspectives (Dal Basco et al., 2015; Mattioli et al., 2016).
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1.3 Process of Sprouting Sprouting is a very easy, low-cost procedure that uses very little room in the greenhouse, doesn’t need sophisticated equipment, and produces high yields. To put it simply sprouting is the beginning of the seed’s transformation into a plant. Through the different metabolites that the parent plant transfers to the seeds, the environment has an indirect impact on this process. There are many steps to the germination process, including enzyme activation by water consumption, the growth of the radicle to act as a root and the growth of the plumule to serve as a shoot. Seeds eventually release enzymes that aid in the breakdown of stored nutrients into simpler molecules and begin to absorb water. At various stages of seed germination, a variety of vitamins, minerals, and enzymes play a crucial role in several ways. Proteins, carbohydrates, phosphate, and lipids that are stored in seeds serve as the carbon framework and a source of energy as a reserve food. The purification, soaking, and sprouting stages of the germination process may be used to categorize the intricate physical and physiological changes that take place. Bacteria can be killed on seeds before sprouting with chemicals like calcium hypochlorite and sodium hypochlorite, which can be contaminated by handling (Sauer & Burroughs, 1986). Soaking the seeds in water or other osmotic solution is the most prevalent method for rehydrating them before germinating. For the growth of germs, soaking is a good option. Therefore, the soaking water should be replaced frequently to prevent germ growth (Idowu et al., 2020). While soaking seeds for a short period of time does nothing to boost their phytochemical content, overdoing it may encourage microbial growth and seed fermentation. To ensure optimal soaking, variables such as the ratio of seed weight to water volume, soaking period, and temperature are tracked. Typically, based on the properties of the seeds, soaking can last up to varying length of time at room temperature (Gan et al., 2017). The ability of soaked seeds to absorb water can be increased by aerating the soaking solution and adding a little salt (Nascimento, 2003). Furthermore, light, temperature, time, and moisture must all be carefully regulated because they have an impact on how seeds behave while sprouting. Typically, for the majority of seed species, sprouting takes place in the dark at a temperature of 10–20 °C (Dove, 2010). The length of time a sprout takes to germinate depends on its use. The time frame between when the seed’s composition significantly changes and when mature sprouts are harvested, varies for the majority of edible seeds and is between 3 and 5 days (Dove, 2010).
1.4 Change in Composition During Sprouting The foremost stage in the multi-phase mechanism of seed germination is for the dry seed to take in water and concludes with the embryonal axis extending (Bewley, 1997). The following biochemical processes are connected to seed germination: (1) using food reserves that have been saved (2) The seed cover softens (3) Activation
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of gene expression and metabolism (4) Organelle rearrangement and biosynthesis (Logan et al., 2001). When a seed is submerged in water, it first takes in a lot of water until all the cells are completely moist, then it takes in less water and starts to burn calories. The germination process is then completed as a result of increased water intake, which causes the cell to lengthen (Bewley, 1997). Numerous primary and secondary metabolites are created and mobilised as a result of the metabolism being reactivated, providing several health advantages that the non-sprouted seeds do not. When it comes to their biochemical makeup, seeds are heterogeneous. The biochemical composition of the seeds has an influence on how well they germinate. Studies on the mobilisation of seed reserves during sprouting have been conducted in a variety of ways (Galland et al., 2017; Yang et al., 2007; Rosental et al., 2014; Sreenivasulu et al., 2008).
1.4.1 Carbohydrates Starch mobilisation is a well-studied mechanism in seedling growth, the considerable changes in grain carbohydrates in most sprouted grains have been well explored (Aoki et al., 2006; Gujjaiah & Kumari, 2013). Amylases catalyse the hydrolysis of starch to produce glucose and maltose, the grains’ amylose and amylopectin (which are stored in the grains) become more easily digestible (Chung et al., 2012; You et al., 2016). It is anticipated that soluble sugars would be used in substantial quantities during respiration as germination develops. Another problem is that there isn’t enough α-amylase in the system to hydrolyze and release sugars from starch (Mbithi- Mwikya et al., 2000). As amylolytic enzymes produced in the aleurone layer permeate the endosperm, dormancy is broken after 36–48 h of germination (Mbithi-Mwikya et al., 2000). When the seeds germinate, the soluble sugars glucose and fructose, which are produced as a result of sucrose hydrolysis by the enzyme invertase, expand considerably as a result of the enzyme’s activation (Traore et al., 2004). Species-specific changes in the sugar profiles of germinating grains are observed. For instance, buckwheat stores more glucose than maltose, whereas rice, sorghum, and millet do the reverse (Agu et al., 2012; Chiba et al., 2012). Soluble sugars rise during germination, and starch is the primary metabolite needed by the seed as it intakes water from the environment (Zhao et al., 2018). There is a correlation between the amount of seed reserves mobilized and quantity of it present in seed and also depends upon the plant species. Water-insoluble cellulose, hemicellulose, and lignin, as well as β-glucans and arabinoxylans (water soluble) are the structural carbohydrates present in the seed (Hübner & Arendt, 2013). β-glucans are β-D-glucose linear polysaccharides having 1–3 β-glycosidic bonds. Arabinoxylans are soluble non- starch polysaccharide found in the cell wall, attached to the ferulic and p-coumaric acids via ester linkage.
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Sprouting can have a variety of effects on non-structural carbohydartes, depending on a number of variables, including fibre percentage, germination duration, and genotype (Nelson et al., 2013). In the first 48 h of incubation, the dietary fibre content of germinating wheat declined, but it increased after 196 h of incubation (Koehler et al., 2007; Hung et al., 2012). When it came to barley, however, the incubation period of 72 h revealed no discernible difference (Teixeira et al., 2016). There was a massive increase in non-structural carbohydrates during rice malting because new main cell walls were formed (Lee et al., 2007).
1.4.2 Proteins The alcohol-soluble prolamins, alkali-soluble glutelins, salt-soluble globulins, and water-soluble albumins are the key storage proteins observed in most cereals. After 2–3 days of imbibition during grain germination, proteolytic enzymes hydrolyze the store proteins into peptides and amino acids, enhancing the bioavailability of the nutrients (Taylor et al., 1985). Germination appears to have a varying impact on protein. Grain and seed protein content has been demonstrated to increase following germination in diverse ways (Laxmi et al., 2015; Otutu et al., 2014). As quinoa germinates, a rise in tryptophan and lysine amount, has been seen in several studies, whereas others have seen a reduction in total protein (Bhathal & Kaur, 2015). It is possible that the increase in proteins is due to the breakdown of lipids and carbohydrates during respiration and the formation of amino acids in the course of germination (Ongol et al., 2013; Jan et al., 2016). Proteases are also thought to be involved in protein degradation during germination. Wheat, barley, rye, and other cereal grains (Lorenz & D’Appolonia, 1980) and triticale, for example, have all been shown to lose prolamins as sprouting time rises (Koehler et al., 2007). The cereal’s proteins and amino acids break down into transportable amides as a result of water absorption, which are then delivered to the developing sections of the seedlings. This is contrary to the findings of many authors, who have noted a rise in crude proteins in barley, Brown rice (Moongngarm & Saetung, 2010), waxy wheat (Hung et al., 2012), and oats (Tian et al., 2010; Singkhornart & Ryu, 2011). During germination, however, the balance of protein synthesis to protein breakdown controls the protein content (Megat Rusydi et al., 2011). 1.4.2.1 γ-Aminobutyric Acid GABA is a non-protein amino acid. Its precursor is L-glutamine. It acts as an inhibitory neurotransmitter. There has been extensive research on GABA generation in brown rice, but wheat and barley germination have also been found to have variations in GABA concentration (Hung et al., 2012; Chung et al., 2009). During sprouting, GABA levels rise rapidly, regardless of species. The content of GABA increases
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2–5 times in brown rice seedlings depending on the varieties (Yao et al., 2008), and when sprouted for 2–4 days at 27–35 °C, it increased by up to 8- to 12-fold (Roohinejad et al., 2011; Cornejo et al., 2015). A reduction in glutamate content and elevated glutamate decarboxylase (GAD) activity made this possible. Steeping or soaking of grains prior to germination and environmental factors both have a substantial impact on the GABA content in cereal seedlings.
1.4.3 Lipids In cereals, lipids in the form of oil (triacylglycerols, TAG) are prevalent in the aleurone, scutellum, and embryo. The net conversion of oil to sugars that occurs during germination is required for the release of TAG from oil bodies. The initial release of esterified fatty acids from TAG is caused by lipases. It is possible to turn free lipid acids into sugars once they have been degraded by glyoxylate and oxidation cycles (Graham, 2008). The mentioned literature can be utilised to understand more about how TAG is converted into sugars during seed germination. Among cereals, oats are distinctive due to their high oil content relative to starch and protein concentration, were observed to break down their oil reserves from the embryo before those in the scutellum. After 1–2 days of imbibition, the TAG reserves in endosperm began to mobilise. This process happened at the same time when this tissue began to accumulate free fatty acids (Leonova et al., 2010). Not only did the number and composition of key FAs (Linoleic and linolenic acids) in the first 48 h of sprouting remain unchanged, but so did the proportion of both unbound and bound lipids (Hung et al., 2012). Linolenic acid was significantly more abundant in wheat seedlings that had sprouted for 9 days, although cis and trans oleic acid and trans linoleic acid concentration was lower. The most prevalent Fatty acids in wheat sprouts after 3 days of germination were oleic acid, palmitic acid, and linoleic acid (Marton et al., 2010). Furthermore, in this case, the metabolic dynamics of FAs are significantly influenced by treatments applied prior to germination and activity of lipase in whole grain tissues, with certain FA levels either increasing or decreasing during the course of germination.
1.4.4 Phytate and Minerals In many plant-based foods, such as mature grains and legumes, phytate is highly concentrated and serves as the primary phosphorus storage form (Kumar et al., 2010). Due to a lack of intestinal phytase, the bioavailability of Ca2+, Mg2+, Mn2+, Zn2+, Fe2+/3+, and Cu2+in humans is adversely affected by phytate, which chelates cations with a high affinity (Azeke et al., 2011).
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Phytases are histidine acid phosphatases with a large molecular weight that hydrolyze phytate to produce myo-inositol, orthophosphate, and inorganic phosphate. The phytase activity often increases throughout germination. In the case of barley, Sung et al. (2005) discovered that phytase activity was extremely low at the start of sprouting but climbed to an eightfold level within the first few days. Whole grain phytase concentrations differ significantly among cereal species, with Oats possessing the least amount and rye has the highest (Hübner & Arendt, 2013). Phytate concentration therefore decreases during germination to varying degrees (Azeke et al., 2011). In case of brown rice, the phytate content was reduced to 60% by carrying out the sprouting for 12–72 h (Liang et al., 2007), whereas in sorghum sprouting for 4 days reduced the phytate content up to 87% (Maghoub & Elhag, 1998). Minerals and phosphorus are more bioavailable as phytate level is reduced. In response to germination, wheat and barley both experienced a decrease in Ca concentration and an increase in Mg concentration (Plaza et al., 2003). Following 96 h of germination, the Ca (76.9%), Fe (18.1%), and Zn (65.3%) extractability from the whole grain of finger millet increased to 90.2%, 37.3%, and 85.8%, respectively (Mbithi-Mwikya et al., 2000). Lemmens et al. (2018) found that after sprouting, bio-accessibility of Zn and Fe rose from 15% to 27% and 14% to 37%, respectively, when wheat was hydrothermally treated.
1.4.5 Antioxidants Polyphenols, carotenoids, ascorbic acid, and tocopherols are antioxidants that protect seedling cell components from oxidative damage and are abundant in whole grains (Bailly, 2004). Phenolic acids, both unbound and bound, can be found in seeds, with bound forms predominating. The hydrolyzable tannins, lignins, cellulose, and proteins that make up bran and aleurone are related to these bound components (Engert et al., 2011). During germination, the total amount of polyphenols increases somewhat, Despite the fact that unbound and bound fraction contributions vary based on the species and sprouting conditions (Alvarez-Jubete et al., 2010b; Pal et al., 2016). When wheat is sprouted under controlled conditions, the unbound fraction grows after 2 days, whereas the bound fraction shrinks (Van Hung et al., 2011) and same is observed in seedlings of emmer or einkorn that have been grown for 12 days (Van Hung et al., 2011; Benincasa et al., 2015). For the flavonoids, Quercetin, catechin, and rutin in tartary buckwheat sprouts had higher concentrations than their whole grains (Pongrac et al., 2016). The β-carotene levels in wheat increased during germination, whereas its amount is variety dependent in barley malts, either showed slight increases or decreases (Yang et al., 2001; Goupy et al., 1999). It is likely due to the de novo production of vitamin C that germinated wheat and barley have higher vitamin C contents (Plaza et al., 2003; Yang et al., 2001; Lemmens et al., 2018), notwithstanding the fact that
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vitamin C concentration in grains is typically relatively low. Furthermore, it was also shown that rice that had been germinated had a much greater tocopherol concentration (Lee et al., 2007) because tocopherol is crucial for preventing non- enzymatic lipid oxidation when seedlings are growing.
1.5 Factors Affecting Nutritional Quality During Sprouting 1.5.1 Genotype and Seed Source The genotype plays the most important part in determining the nutritional content of grains that have been sprouted. Recent studies have focused on defining grains from various historical and contemporary cereal genotypes, as well as pseudocereal taxa, primarily in terms of their bioactive constituents. The environment has a well-known impact on whole grain biochemical composition during crop growth, particularly during development of grain. Research by Bellato et al. (2013) evaluated the polyphenol content, anti-radical activity, and 5-n alkylresorcinol concentration of 30 commercial durum wheat varieties grown in two distinct geographic locations in Central and Southern Italy. Overall variability was shown to be less dependent on the genotype-by-environment interaction (GxE) than independent outcomes (G and E), with environment accounting for the largest share of variance. Dry conditions during filling of grains increased alkylresorcinol concentrations, while high availability of water during grain growth favoured the accumulation of free phenols. The environment has also been found as the principal driver of overall variance in various quality parameters for winter and spring wheat types (Shewry et al., 2010). Wheat grain nutrition is also affected by the mother plant’s vulnerability to biotic stress. Numerous studies emphasising on comparing the nutritional value of organic and conventional cereals have produced conflicting findings (Mazzoncini et al., 2015). Older wheat varieties have greater nutrient utilisation in low-N conditions than modern wheat varieties, which solely relyon high amounts of available N (Di Silvestro et al., 2012). However, protein content is generally lower in organic commodities. Because phenolic compounds tend to accumulate under biotic stress conditions, it is often assumed that organic crops have larger quantities of secondary compounds. However, even for specific phenolic acids, the impact of the cultivation strategy on the concentration of secondary metabolites is typically negligible. Co-variables naturally alter the impact of various agronomic techniques and environmental conditions on plant secondary metabolites. Three Chilean landraces of quinoa exposed to 100 and 300 mM NaCl starting 34 days after planting showed significant changes in the protein profiles and amino acid composition of main seed storage proteins and bioactive compounds (Aloisi et al., 2016).
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1.5.2 Germination Conditions Grain “seed invigoration” treatments, as well as the germination conditions, both influence the biochemical changes that take place during sprouting, as well as subsequent seedling growth. During seed priming, Seeds are hydrated with a solution that promotes imbibition and the start of the first reversible germination stage but inhibits radicle protrusion (Lutts et al., 2016). There are three regularly utilised priming methods: osmo-, halo-, and hydropriming. Various osmotic solutions are used for soaking seeds such aspolyethylene glycol (osmopriming), salt solution (halopriming) and water (hydropriming). Researchers have recently concentrated their efforts on determining the ideal pre-sowing and sprouting conditions in terms of temperature and timing to produce sprouts with a high phytochemical content. Unfavorable germination conditions may lead seedlings to accumulate phytochemicals because of the stimulation of secondary metabolism (Liu et al., 2019). Rehydration causes significant levels of oxidative stress, consequently there is rise in the ROS formation which may harm the biomolecules in the seeds. Therefore, scavenging of ROS is crucial, done by non-enzymatic antioxidants (e.g. phenolics) and their amount rises when seeds germinate under stress conditions (Tan et al., 2013). The stress which a seed grain faces during the pre- treatments and sprouting should be highlighted because they may lower the percentage of germination and/ or the amount of dry matter produced. Therefore, in order to create sprouts with superior nutritional properties without affecting their yield, the commercial uses of manipulating germination parameters should be properly regulated for each species. 1.5.2.1 Temperature Most of the research has focused on non-graminous plants, such as lentil (Świeca & Baraniak, 2014), broccoli (Guo et al., 2016a), alfalfa, and radish (Oh & Rajashekar, 2009), rather than graminous plants like corn or wheat. GABA content rose after 72 h of steeping hullless barley sprouts in water at 5 °C (Chung et al., 2009). A 48-h period of germination followed by anaerobic and heat treatments, the GABA content of dark northern spring wheat and soft white winter wheat increased (Youn et al., 2011). The development of anthocyanin in a variety of plants is influenced by cold stress, including 12-day-old tartary seedlings cultivated at 4 °C for 4 days (Li et al., 2015). As a result of the enhanced anthocyanin content in stressed buckwheat sprouts, ROS were reduced and cell osmotic potential was decreased (Nagata et al., 2003). In addition, the more bright colours of anthocyanin-rich sprouts help the enhancement of their nutritional and health properties to attract more customers.
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1.5.2.2 Light Modulations Light is momentous for proper growth of the plant. Plant morphogenesis, the efficiency of the photosynthetic system, and the progression of metabolic pathways are all influenced by how photoreceptors react to variations in light intensity, quality, duration, and intermittency (Samuoliene et al., 2011). Additionally, variations in illumination could cause photoxidative alterations in plants, which modify how the mechanism of antioxidant defence functions (Samuoliene et al., 2011). Although the impacts of light intensity are widely known, there is currently little information on how light spectrum quality affects plant metabolism. In terms of sprout production, the most effective method to enhance the seedlings nutrient content appears to be the employment of light emitting diodes (LEDs) during seed germination. There are various ranges of LEDs from the near ultraviolet (UV) to the near infrared (IR), each having a distinct wavelength characteristic. Thus, in order to enhance the concentration of particular molecules, light spectra may be produced by choosing particular wavelengths and combining them. Various LED treatments have been studied to see how they affect pigment concentrations and the carbon-nitrogen metabolism. Blue light boosted the chlorophyll a/b ratio, total proteins, and chlorophyll fluorescence in the leaves of two rice cultivars with 14-day-old seedlings (purple and green leaf), whereas red + blue LEDs had the maximum anthocyanin content (Chen et al., 2014). Red and blue LED-grown barley seedlings have twice as much amino acid content as control plants cultivated in sunlight (Meng et al., 2015). Tartary buckwheat sprouts exposed to white light rather than blue or red light showed the most increase in total carotenoids (Tuan et al., 2013). In common and tartarian buckwheat, increasing daytime light periods resulted in higher contents of rutin, free amino acids, and vitamin C than growing under complete darkness (Kim et al., 2006). The total phenolic and flavonoid content of 7-day- old buckwheat sprouts grown in a dark environment, were higher than those grown in an open field in a pristine environment (Sharma et al., 2012). But in the earlier study, the natural light intervals did not last all the time during sprouting periods, and the germination conditions for other factors in the subsequent study varied. Additionally, sprouts that were grown in the light for a longer period of time (14 days) contained substantially more rutin and total flavonoids than sprouts that were grown in the dark (Yao et al., 2004). This might be accounted for by the fact that many structural genes engaged in the formation of flavonoids are activated in the presence of light, as evidenced by Tartar buckwheat sprouts that are 2, 4, and 6 days old in a light and dark environment versus those that were only exposed to darkness (Li et al., 2012). The duration, spectrum, and intensity of the light also have an impact on energy consumption (Tosti et al., 2018) and, consequently, Noteworthy is the price of growing indoors, which is the quite frequent circumstances in sprout production.
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1.5.2.3 Salt Stress Among the most significant abiotic factors that can put plants under stress is salinity, particularly early on in the process of a seedling’s development. But as of now, little is understood about how salinity affects the accumulation of phytochemicals in edible sprouts since investigations on the effects of high salt intake have mostly focused on germination rates and physiology. Since there haven’t been many studies on grains, data from some non-gramineous species is necessary for a thorough examination of the literature that is currently available on this subject. Treatments with NaCl solutions (10, 50, 100, and 200 mM) raised the number of carotenoids and phenolic compounds, including isoorientin, orientin, rutin, and vitexin in buckwheat sprouts regardless of growth stage (Lim et al., 2012). The total bound-phenolics percentage in einkorn rose up, until 50 mM NaCl solution, but durum wheat responded to salt by raising the overall amount of free phenolics while lowering the binders (Stagnari et al., 2017). Einkorn and emmer wheatgrass aqueous raw and denatured extracts were assessed for their total phenolic content, superoxide radical scavenging, reducing power, and ability to prevent the formation of reactive chemicals that react with thiobarbituric acid. It was found that for these genotypes, 25–50 mM NaCl provided the ideal balance between growth and quality features, which maximised polyphenol and antioxidant output (Falcinelli et al., 2017). The maximum GABA buildup was seen in seedlings of 5-day-old tartary buckwheat in a 34 mM NaCl solution (Zhu & Guo, 2015). 1.5.2.4 Hypoxia Stress During germination, a variety of species have been investigated to see how the composition of gases changes. As a result of hypoxic stress while sprouting, soybean (Guo et al., 2011, 2012), faba bean (Yang et al., 2013) and brown rice (Ding et al., 2016) seedlings accumulated GABA. Increased GABA synthesis is one way that plants respond to oxygen scarcity stress-induced cytosolic acidosis (Aurisano et al., 1995). In fact, hypoxic stress and acidic culture solutions as germination substrates are widely researched together. Under hypoxia, sprouted foxtail millet had the maximum GABA concentration when germinated at 33 °C with 1.9 L/min of air flow and a pH solution of 5.8 (Bai et al., 2008). An acetate buffer solution was found to be less effective than a citrate buffer solution (10 mmol/L). Tartary buckwheat germination at 31.25 °C, 1.04 L/min air flow, and 4.21 pH (citric acid buffer – 10 mmol/L) yielded the best GABA concentration, according to Guo et al. (2016b).
1.5.3 Biofortification Food biofortification programmes could use germinated whole grains as a delivery method. Wei et al. (2013) discovered that pre-germination soaking of brown rice kernels in FeSO4 solutions increased the Fe content in grains germinated for 24 h in a
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recent study. Fortification of brown rice sprouts with ferrous iron (Fe) raised Fe content by 1.1–15.6 times owing to Fe solutions accessing aleurone layers across the endosperm’s dorsal vascular bundle (Fukai et al., 2008; Wei et al., 2013). However, the comparatively low permeability of certain seed coverings prevents fortification with solutions that are high in iron. This was the situation with broccoli and radish that had been immersed in solutions of Fe(III)-EDTA and Fe(III)-citrate (Park et al., 2014). In the course of germination, seeds have the potential to store selenium (Se) and convert it from an inorganic to an organic form (i.e., proteins incorporating selenium). In order to boost the concentration of Se in seedlings, Se-biofortification during sprouting may be an effective technique. To test this hypothesis, the Se concentration in buckwheat tartary sprouts that are 3 days old, increased as the amount of external selenite treatments increased. The amount of selenite found in brown rice seedlings increased dramatically with both the amount of exogenous selenite present (up to 60 mole/L Na2SeO3) and the duration of time since germination (4-day old sprouts). Although the distribution of Se-containing proteins was found to be significantly uneven, germination time increased the conversion of inorganic selenium to protein-bound selenium (Liu et al., 2011). Germination media containing Na2SeO3 over a period of 24 h at 25 °C (Lazo-Vélez et al., 2016) can be used to create Se-enriched wheat kernels, along with some enzyme and performance properties (such as -amylase activity). Implementing fortification programmes during crop cycles is a successful method in order to increase the amount of macro- and microelements present in whole grains and to have an effect on their dynamics throughout the germination process that follows. An efficient agronomic technique to boost the Zn content in rice grains is foliar Zn fertilisation, which is used over the phases of the formation of the panicle and the filling of the grain (Wei et al., 2012). When Zn-containing urea was applied, maize grains’ levels of Zn and protein rose, producing better outcomes in soils with low Zn content (Messias et al., 2015). Common buckwheat’s ability to absorb selenium was greatly improved by foliar and soil applications of selenium, with soil applications of selenium exhibiting the strongest association with grain selenium concentration (Jiang et al., 2015).
1.6 Benefits of Sprouts 1.6.1 Antioxidant Activity Studies on plant extracts ability to neutralise free radicals have been a key focus. It has been found that sprouts are rich in antioxidants, both phenolic and non-phenolic. Ascorbic acid has been studied in sprouts (Hamilton & Vanderstoep, 1979). Glucosinolates found in broccoli and cabbage are capable of scavenging reactive oxygen species (ROS). It has so become increasingly popular to eat sprouts as a result of their useful qualities (Randhir et al., 2008; Majid & Nanda, 2018; Majid et al., 2016).
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1.6.2 Cytotoxic Activity Chemicals and nanoparticles are ubiquitous in our daily lives, therefore human exposure to them is unavoidable. It has been demonstrated that certain chemicals and nanoparticles severely hamper living cells. As a result of increased exposure to carcinogens, cancer has recently become a major public health issue. About 599,000 Americans lost their lives to cancer in just 1 year (Centers for Disease Control, Prevention, 2005). Scientists have investigated whether plant sprouts could be used to cure cancer because of the high mortality rate among cancer patients. On the other hand, a study conducted by Gawlik-Dziki and his colleagues found that sprouts had a strong inhibitory effect on prostate cancer progression (Gawlik-Dziki et al., 2012). Young red cabbage shoots have been shown to have greater anticancer action than mature veggies because of their high amount of Glucosinolates (Drozdowska et al., 2020). Buckwheat sprouts have been shown to have anticancer properties due to the modification in gene expression mediated by active chemicals in the grain (Gimenez & Zielinski, 2015). Rutin and quercetin, two phenolic components found in buckwheat, can annihilate cancer cells, block the cell cycle, and limit the proliferation of malignant cells (Guo et al., 2010; Sak, 2014).
1.6.3 Antidiabetic Activity Blood sugar levels in people with diabetes mellitus are abnormally high (hyperglycemia). Both defects in insulin secretion and actions can contribute to hyperglycemia, and in some situations, both can happen at the same time (Association, 2010). Seeking antagonists to halt or slow the hydrolysis of carbohydrates by enzymes like alpha-glucosidases, which prevents the buildup of sugar, is a major focus of research into treating diabetes (Bao et al., 2016). Researchers discovered that the natural antioxidants found in the sprouts of the majority of plant species can protect the body from the detrimental outcomes of oxidative stress and halt the degradation of complex carbohydrates into sugars.
1.6.4 Hypocholesterolemic and Anti-obesity Activity The body’s oxidative stress may be exacerbated by an increase in cholesterol intake, which could lead to an increase in LDL and LDL’s oxidised form (oxLDL). As a result, atherosclerosis and other heart-related conditions may develop (Gimenez & Zielinski, 2015). Research shows that sprouted seeds can protect against heart disease caused by an imbalance in cholesterol levels, both in vitro and in vivo. Buckwheat sprouts and seeds were studied by Lin et al. (2008) for their hypolipidemic properties. Studies
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found that buckwheat meals (seed and sprout meals) reduced LDL cholesterol levels considerably, indicating that the sprouts have an inhibitory effect on hypolipidemia (Lin et al., 2008). Alfalfa sprouts, however, have been allied to a decline in blood plasma cholesterol levels and a blocking effect on absorption (Harwood et al., 1993; Petit et al., 1995). Alfalfa saponins have been shown to increase the liver’s ability to turn cholesterol from the liver into bile salts, which leads to the elimination of the salts.
1.6.5 Antiviral Activity Globally, viral infections are a leading cause of death. HIV, hepatitis B and C, and influenza (Neagu et al., 2018) all have antiviral treatments that have been discovered in the past. Drug-resistant virus strains have emerged as a result of their widespread therapeutic use. In immunocompromised patients, the harmful effects of several antivirals hinder efforts to find a cure for viruses. In response to these challenges, scientists have stepped up their efforts to produce antiviral medicines from plant bioactive compounds (Hafidh et al., 2015). Influenza virus-induced markers of inflammation can be improved by consuming a small amount of broccoli for a few days (Noah et al., 2014). In addition, sprouts from mung beans have been shown to lessen the risk of viral infection. Antiviral phytoagents could be developed by comparing the effectiveness of various sprouts or sprout extracts against viruses. By finding antiviral medications from these extracts, drug-resistant viruses could be avoided.
1.7 Nutritional and Food Safety Issues Seed germination facilitates digestion and absorption of nutrients by breaking down lipids and proteins. Before being placed in a warm, humid environment for germination, seeds must first be soaked in water. Bacterial populations can flourish in these circumstances. Most contamination occurs during germination, when bacteria from seeds are consumed by sprouts and spread to other foods (US FDA, 1999). Studies have found that sprouts had counts that were 2 or 3 logs higher than seeds, which had counts between 3.0 and 6.0 log CFU/g. Germination is just one of several conceivable points at which sprouts can become contaminated. Some types of contamination are referred to as “pre- or post-harvest contamination,” depending on when they occur. The type of fertiliser used, the water utilised for irrigation, and the state of the soil are all potential pre-harvest contamination sources. Because of how they are handled and stored throughout transport, shipping, and storage, sprouts might get infected after harvest. Preserving the health benefits of sprouts by eating them raw is a common practise in Western countries. No procedures are taken to
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ensure that any harmful bacteria contained in sprouts have been removed before to ingestion, making sprout eating unsafe. The Food and Drug Administration (FDA) of the United States has issued a number of consumer-friendly guidelines on sprout eating (US FDA, 1999, 2012). Consumers are advised to boil the sprouts sufficiently in order to minimise or eliminate the microbial load, as cleaning the sprouts before to eating is useless. Additionally, approaches for decreasing or eradicating bacterial populations in sprouts that have been contaminated before to or after harvest have been investigated. Physical, biological, and pharmacological treatments are all on the list. Physical treatment involves heat (Jaquette et al., 1996), cooling (Tian et al., 2012), high pressure, irradiation, and supercritical carbon dioxide. Antibiotic metabolites, antagonistic bacteria, and bacteriophages make up biological treatments (Liao, 2008; Ye et al., 2010). An electrolyzed water treatment is a chemical intervention, which includes ozone (Singla et al., 2011), as well as chlorine (Bang et al., 2011). Research have shown that seeds may be infected with up to 106 CFU/g of bacteria (Zhang et al., 2011; Prokopowich & Blank, 1991; Robertson et al., 2002).
1.7.1 Preharvest Sources of Contamination There are two key elements that influence contamination of the final product: pathogen load and seed quality. Only a small percentage of the harvested seeds are used for sprouting, which is mostly for agricultural purposes. Harvest is typically the last stage before deciding whether or not to use the produce. Some seed growers ignore good agricultural practises (GAPs) because the seed is considered an agricultural product instead of a food (NACMCF, 1999; Robertson and others, 2002). Pathogens can enter a seed in a variety of ways. This includes manure, which is one of the sources used in fertiliser (CFIA, 2007). A cheap and efficient technique to raise the quality of agricultural soil is by adding animal manure. Campylobacter spp. and Salmonella spp. can be found in animal manure (Ostling & Lindgren, 1991; Guan & Holley, 2003). As a result, adding manure to the soil may introduce these pathogens. After then, the viruses have a chance to attach to the fruits and vegetables. Fimbriae, flagella and biofilms can all be used to attach foodborne pathogens to surfaces. Jeter and Matthysse discovered in a 2005 investigation that E. coli O157:H7 strains, but not other coli bacteria, were able to cling to alfalfa sprouts. More than one method for adhering to plant surfaces was suggested in their research. As a result, understanding how viruses attach to seeds and sprouts is critical if we are to avoid their spread. In addition, it is possible that dangerous bacteria can survive in seed fractures and cavities, indicating that seeds should be protected from damage. It is also possible that contamination could come from the place where the crops are grown. Fertilized grasslands and pastures are more prone to be tainted by harmful bacteria than are other types of fields because of the presence of animal faeces
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(Brackett, 1999). There is a chance that bacteria like Salmonella and E. coli O157:H7 to linger for several months in faeces or soil. As a result, contaminated fields would serve as a long-term reservoir for infections, contaminating the sprouts that grew there. Irrigation systems are another major source of contamination, as is their source of irrigation water. Due to a paucity of high-quality water, low-quality water is frequently used. Potable water, rainfall, groundwater, surface water, and effluent that hasn’t been properly treated are all sources of water that can be used for irrigation (Pachepsky et al., 2011). Because of this, the latter is more prone to infection by disease. The microbiological load of water has been established in a number of studies. According to studies by Cross (1997), there was an 8% risk of faecal coliforms in irrigation water samples in Alberta, Canada. In another investigation, two of the 96 water samples obtained from a river in northern Nigeria that is extensively used for irrigation contained E. coli O157 (Chigor et al., 2010). Johnson and his coworkers claim that Salmonella species and E. coli O157:H7 were found in southern Albertan water samples at rates of 0.9% and 6.2%, respectively (Johnson et al., 2003). According to these investigations, the quality of the cultured product can be negatively impacted by microorganisms in irrigation water. Direct contact with infectious farm workers might also contaminate seedlings. Workers in the field should have access to clean and sanitary restrooms, as well as proper hand-washing facilities. Workers should also acquire the appropriate hygiene training for their profession.
1.7.2 Postharvest Sources of Contamination Seeds and sprouts come in contact with debris and water during harvest, increasing the risk of infection. Furthermore, they are not separated until they are delivered. This means that the source of contamination must also be assessed in light of the presence of contaminated harvesting equipment, storage facilities, and transportation vessels. Because of the high level of human contact during harvest and post- harvest processing, employees with subpar personal hygiene are a source of infection. Food safety is a subject that many individuals lack even a basic awareness of. In a research on safety evaluations, 23 of the 45 enterprises examined (NACMCF, 1999) lacked hot water for cleaning. The sprouts are susceptible to various sources of contamination once they have been removed from the sprouting site. For instance, sprouts can become contaminated in eateries like restaurants and delis as well as the customer’s home. A lack of proper storage conditions encourages the growth of microorganisms. Using infected food handlers or eating at places with low hygiene standards can also spread germs into the food.
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1.7.3 Factors Affecting the Survival and Growth of Pathogens The pathogen’s infectious dosage and survival rate during storage determine the relevance of product contamination. Several factors, both inherent and extrinsic, influence the growth of bacteria in sprouts while they are being stored. Among them is the temperature at which products are kept in storage. Castro-Rosas and Escartin (2000) investigated the effects of refrigeration on pathogen viability. The amount of viable S. typhi was observed to be decreased by cooling after 15 days of storage by 50% whereas Vibrio cholera concentration reduced by 90%. It can be extrapolated from this finding that refrigerated sprout storage can lower the danger of acquiring a foodborne illness. Pathogen expansion is also influenced by factors such as humidity and gas environment composition. Sprouts are sometimes sold unpackaged to the public in open marketplaces. To avoid contamination from the air or from customers who have been ill, sprouts should not be sold this way. Maintaining product quality and microbiological safety require the use of proper packaging conditions. Pathogens’ ability to thrive and grow is also influenced by the availability of nutrients. A considerable rise in water content, which is favourable to bacterial development, was demonstrated by Hamilton and Vanderstoep (1979). It is further claimed by them that the seed and sprouts of alfalfa contain more protein and carbohydrate content than other plants, such as broccoli or lettuce. Sprout is a great substrate for bacterial development because of its approximately neutral pH and the other qualities listed above. Last but not least, bacterial growth on sprouts may be impacted by the development of biofilms, the presence of antimicrobial substances, and competing microorganisms. The high levels of background microflora (108 CFU/g) found in the soybean sprouts tested by Francis and O’Beirne (2001) prevented the expansion of L. monocytogenes. According to Wills and others (1984), organic acids like malic or citric have been found to prevent bacterial development in mung bean sprouts. Antimicrobial compounds in sprouts are often minimal, as evidenced by this finding. Salmonella and E. coli O157:H7 can thrive in the naturally occurring biofilms on sprouts, according to a study by Fett (2000). As a result, foodborne bacteria’ survival and growth on sprouts may be influenced by a variety of variables. To limit the risk of foodborne illness related with sprout eating, these parameters must be controlled to suppress growth of pathogens.
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Chapter 2
Barley Sprouts Mamta Thakur
and Sudha Rana
2.1 Introduction Consumer lifestyles today have changed to emphasize “healthier living and healthier eating,” and as a result, the demand is more focused on nutrient-dense, fresh, and wholesome foods having high concentrations of bioactive substances. Barley, scientifically known as Hordeum vulgare L. is a historical cereal grain that has long been utilized as a primary ingredient in the breweries (Kok et al., 2019; di Vaio et al., 2023). Along with proteins, the high starch content of barley which is employed for malt preparation and in brewing process, normally provides more than enough amino acids for yeast development and nitrogenous substances, which are crucial for producing beer foam. Barley outperforms almost all other cereal crops in respect of drought tolerance. Having higher nutritional value and amount of physiologically active substances, barley continues to be a significant grain in various Asian and Northern African civilizations today. Barley has primarily been utilised for animal feed composition, 1/3rd for malting, but very low amount i.e. 2% as a direct food ingredient, according to di Vaio et al. (2023). Barley seeds’ medicinal and nutritional benefits are further increased during sprouting. The result leads to higher levels of simple sugars, free amino acids, and organic acids along with the catabolism and breakdown of the primary macronutrients of barley, such as carbohydrates, protein, and fatty acids (Farooqui et al., 2018; Ortiz et al., 2021). On the contrary, it increases some M. Thakur (*) Department of Food Processing Technology, College of Dairy and Food Technology (Rajasthan University of Veterinary & Animal Sciences, Bikaner), Bassi, Jaipur, Rajasthan, India S. Rana Department of Food Science and Technology, College of Agriculture, Punjab Agriculture University, Ludhiana, Punjab, India © The Author(s), under exclusive license to Springer Nature Switzerland AG 2023 I. Majid et al. (eds.), Advances in Plant Sprouts, https://doi.org/10.1007/978-3-031-40916-5_2
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secondary metabolites like vitamin C and polyphenols in sprouts while decreasing anti-nutritional and indigestible elements like protease inhibitors and lectins (Sonia et al., 2009). Barley seeds have also been linked to numerous bioactivities, including anti-inflammatory, antidiabetic, antioxidant and anti-cancerous properties (Kanauchi et al., 2008; Lee et al., 2015; Aborus et al., 2017). As a result, barley sprouts or barley that has been germinated is a popular food in various developing nations and sprouting also increases the utilization of barley seeds as a functional ingredient in food items. The seed, including barley, used in human diets, can be improved in terms of nutrition by sprouting or germination. The malted or sprouted barley grains that still retain all of the original bran, germ, and endosperm are termed as “whole grains”, provided the sprout growth does not surpass kernel length and nutrient levels have should not decreased. The sprouting process that takes place in germinator chambers doesn’t have any detrimental effects on the environment. Due to the intricate biological and metabolic mechanisms that begin when the seed emerges from its latency stage, the biochemical changes in the chemical makeup of the barley take place during sprouting. As sprouting has commenced, the polysaccharides that make up the cell wall are broken down, allowing enzymes to enter the contents of cell. A varying amount of proteins, lipids, and carbohydrates are broken down into simpler and more readily available substances, like free amino acids, free fatty acids, and simple sugars, respectively (Nonagaki et al., 2010; Ortiz et al., 2021). Additionally, the process of sprouting could change the amount of minerals, vitamins, and phytonutrients like enzyme inhibitors, phenolic compounds, and phytic acid in addition to glucosinolates (Fernández-Orozco et al., 2008; Masood et al., 2014). It has been shown that in addition to being a sustainable process, the germination enhances the nutritional value and functional elements levels as well as flavour, digestibility, and accessibility of grains (Gan et al., 2017). However, the degree of nutritional variations changes during sprouting depends on a number of variables, including relative humidity, species and variety, temperature, sunlight, and oxygen access for aerobic respiration (Benincasa et al., 2019). Because of this, optimising the germination parameters is essential for enhancing a grain variety’s nutritional and bioactive qualities. Because NaCl inhibits the ability of barley seeds to germinate and reduces the rate of germination, Abdi et al. (2016) demonstrated that daily germination under NaCl also reduced considerably. Several studies regarding the impact of sprouting on the nutrient makeup of barley grains in the literature specifically consider the grains that were sprouted for malting purposes in the dark for about 48 h (Hoang et al., 2014; Springer & Mornhinweg, 2019; Rico et al., 2020). However, the evidence on modifications in the nutritional composition of barley sprouted in germinator chambers with continuous light is scarce and lacking (Ortiz et al., 2021). Additionally, the minimal processing and chemical or additive- free sprouting seeds have attracted the majority of recent interest. Considering their remarkable qualities, which include a distinctive colour, a robust flavour, and a substantial amount of bioactive compounds, they might be utilised to improve salads’ sensory qualities, garnish a wide range of high-quality foodstuffs, or create functional foods.
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The authors looked for English-language literature on barley sprouts or germination from 2000 to the present using Web of Science and Pubmed. The relevance of germination on the bioactive chemicals and bioactivities of barley has not been highlighted in any recent articles, and there is a lack of an updated study. We therefore summarized the changes in bioactive components and their contents in barley sprouts in this chapter after briefly describing the sprouting process. We next discussed the techno-functional features of barley sprouts also. This chapter also highlighted the health advantages of germinated barley seeds i.e. sprouts, mainly focusing on biological properties.
2.2 Process of Sprouting It has long been recognised that seeds can sprout, particularly in Eastern nations where eating sprouts is a significant part of culinary heritage. Due to consumer demand for exotic healthy meals and nutrition in Western nations starting in the 1980s, the sprouted seed consumption gained appeal for the same. The intricate process of seed germination begins with the dried seeds absorbing water and continues through the lengthening of embryonic axis (Bewley & Black, 2013). Stage I – initial phase includes the dry seed ingestion and the initial plateau period of water imbibition. Stage II – the intermediate phase involves the visible radicle protrusion via seed covering layers and the plateau phase of water intake. Stage III – the seedling development, also known as final phase in which numerous physico-chemical processes, including as the damage of endosperm and testes and leakage of cell- based solutes take place. Further, the different organelles, membranes, and DNA are repaired and the DNA, RNA, and proteins molecules are produced (Ma et al., 2017). The basic following steps can be used to perform the germination: (i) sterilisation (optional), (ii) soaking, and (iii) sprouting. The sprouting techniques for various seeds may differ, although the fundamental guidelines and steps are often constant (Fig. 2.1). Before the seeds are soaked, the sterilisation is carried out to stop the growth of microorganisms. The most widely used sterilising agents for seed germination, as per Limon et al. (2014), include sodium hypochlorite (NaClO) solutions – particularly 0.07% NaClO solution. Sterilization with NaClO solutions is typically done at room temperature for 5–30 min with a seed weight: solution volume ratio of 1:5 or 1:6. Additionally, it has been observed that sterilising seeds with ethanol or 70% ethanol only takes 3 min (Pajak et al., 2014; Wu et al., 2012). The barley seeds were surface-sterilized by Abdi et al. (2016) by keeping them in 10% NaClO solution for 15 min, followed by three washings in sterile distilled water. However, several researches (Guo et al., 2012; Guajardo-Flores et al., 2013) skipped the sterilising phase before soaking the seeds, likely due to concerns about the potentially harmful effects of the sterilisation reagents on the seeds and the safety of the food being consumed. Sterilization is, thus, not required for seed germination; instead, it should rely on the state of seeds, how frequently the water is changed during sprouting, and the objective of germination. However, a few research have
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Fig. 2.1 Illustration of various steps of barley sprouting process
examined the effects of sterilisation on the outcomes of germination (Sen et al., 2013; Munkager et al., 2020). Following this, seeds should be rehydrated by soaking them in water; the soaking time, temperature, and seed weight (g)/water volume (mL) ratio should all be taken into account. Typically, seeds can be steeped for a few hours to 1 day under ambient temperature (20–30 °C), with a seed weight/water volume ratio ranging from 1:1.5 to 1:20. Perveen et al. (2008) discovered that the germination rate for barley steeped in Ca (OH)2, KOH, and Mg(OH)2 solutions was higher (as 60%, 66%, and 62%, respectively) than water (35%). The fundamental qualities of various seeds, such as their propensity to absorb water, the thickness of their seed coats, and their size, should be linked to these variations in seed soaking conditions. After soaking, seeds can be sown by placing them in specialized incubators or germinators. Considerations for seed germination include light, time, temperature, humidity, and watering. Guan et al. (2019) followed 10–25 °C germination temperature and 6–10 h of soaking to produce germinated barley after sprouting seeds for 1–3 days, whereas Abdi et al. (2016) incubated barley seeds in dark at 25 °C for 1 week to induce germination. Sprouting of seeds is frequently carried out in the shade, and the temperature is typically maintained between 20 and 30 °C. Every day during sprouting, seeds should be watered to maintain a moderate level of humidity that will support their growth. Water should also be changed often, perhaps twice daily, to remove the metabolites of germination and to prevent the
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formation of microorganisms. The duration of sprouting depends on the goal of germination. Therefore, germination is a very quick, low-cost, safe, and environmentally beneficial approach to produce the sprouts. Just a few cultivated edible sprouts, including mung bean, soybean, and peanut sprouts, are currently included in human nutrition. The sprouting process/germination of barley seeds can be a significant bio-processing trend to create functional germinated seeds and sprouts due to the culture benefits and concentration of bioactive chemicals and bioactivities during germination.
2.3 Changes in Barley Composition/Properties During Sprouting The chemical make-up (lipids, proteins, ash, carbohydrate, and crude fibre) of barley grain varies greatly (Table 2.1). According to Ortiz et al. (2021), the substantial water intake during seed germination is the reason why the moisture content of sprouted grains was nine times higher than that of raw barley. Since the starch and sucrose during germination are broken down into mono- and disaccharides by enzymes, like amylases or invertases, this process may enable the production of products with lower added sugar contents without reducing sweetness perception and higher concentrations of simple sugars, free amino acids, and peptides that can act as flavour precursors through the aromatic compounds produced in the process (Desai et al., 2010; Pagand et al., 2017). Besides these, there are many changes which occur particularly in barley seed, discussed as below:
2.4 Changes in Carbohydrates About 58–65% (dry weight) of barley is made up of carbohydrates, primarily starch, which serves as a source of stored energy in plants. Starch is a significant industrial ingredient to processed foods and a major source of calories for the human diet. According to Contreras-Jiménez et al. (2019), the starch has a granular structure composed of amylose and amylopectin. The amount of total starch in barley that has been sprouted for 4 days at 17 °C reduces during the process (Vinje et al., 2015). Similarly, Ortiz et al. (2021) reported that the starch content of barley sprouts was significantly lower than that of native grains, falling by 18.50%.This decrease in starch can be caused its breakdown during sprouting and early growth due to a rise in the function of starch-degrading enzymes. When grains are steeped and sprouted, enzymes such as ß-amyloglucosidase, ß-amylase, α-glucosidase, and α-amylase, digest the starch molecules into smaller compounds, primarily sugars, which the embryo utilises as an energy source (Devi et al., 2015; Sharma et al., 2016). The
Sample Naked barley (cv. Beiqing No.7)
Changes in Sprouting Proteins and conditions amino acids Germination of – naked barley at 25 °C for 12, 24, and 36 h after sterilized using 0.3% NaClO solution for 20 min and then dried using infrared (600 W/m2, inlet air – 20 °C and flow velocity was 0.5 m/s) or hot air (60 °C and 0.01 m/s air velocity) Carbohydrates and fibre Lipids – –
Table 2.1 Changes in barley composition during sprouting Vitamins and minerals –
Phytochemicals References 76 phenolic compounds Ge et al. were found in control (2021) sample of which 33, and 36 were detected for infra-red and hot air dried samples; total phenolic content (TPC) higher in dried samples than control
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Barley (Hordeum vulgare L.)
Cleaned and washed with 0.07% sodium hypochlorite solution. Sprouting was done at 20 °C temp., 80% relative humidity for 6 days
Sprouting Sample conditions Grains soaked in Egyptian barely (Giza water for 12 h and germinated for 126 and 3–4 days and then Giza 132) dried in air for 48 h
Crude protein increased by 38.02% compared to control
Changes in Proteins and amino acids Higher protein levels (12.81%) in germinated barely than regular barley (9.91%); germinated barely showed higher value of leucine, lysine, methionine and phenylalanine
Starch decreased by 18.51% cellulose increases by 248.86% β-glucan decreased by 49.1%
Carbohydrates and fibre Rise in crude fibre content from 4.75% to 5.15% after germination; higher β-glucan in germinated sample
Crude fats increased by 42.04% compared to control
Lipids Lower fat content (2.10%) in germinated barely than regular barley (2.53%)
Vitamins and minerals Decrease in ash content from 3.24% to 2.81% after germination; increase in Ca, Mg, K,Cr and Mn of germinated sample but Fe showed reduction by 17.4% after germination; Rise in folic acid (by 627.5%) and pyrodoxine after germination but vitamin E decreased from 5.66% to 4.89% after germination – –
(continued)
Ortiz et al. (2021)
Phytochemicals References Rise in DPPH antioxidant Lotfy et al. (2021) activity from 56.60% to 82.12% after germination; higher total flavonoids in germinated barley (102.0 mg/100 g) than control (54.38 mg/100 g); higher TPC in germinated barley (86.1 mg/100 g) than 76.5 mg/100 g in normal barely; tannins decreased from 63.46 mg/100 g to 28.20 after germination
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Barley (hull-less H13)
Sample Highland barley
Sprouting conditions Pre-cleaning of barley grains was done using 0.1% sodium hypochlorite. After being cleaned, the grains were soaked in de-ionized water for 24 h at 25 °C. The process of germination took place for 1, 2 and 3 days in the germinator, at 80–85% RH and 30 °C. After soaking barley seeds in 0.1% NaClO for 30 min; seeds were steeped in water for 4 h; then germination was carried in darkness at 12.1–19.9 °C for 1.6–6.19 days with >90% RH
Table 2.1 (continued)
Rise in protein content significantly after germination in all sprouts besides the germination conditions used
Changes in Proteins and amino acids Crude protein decreased by 2.94% on 1st day of germination
Drop in crude fiber in germinated barley than regular barley; reduction in carbohydrate content of sprouted barley at low temperatures and short germination times but remained constant at other temperatures
Carbohydrates and fibre Starch decreased by 11.18% on 1st day of germination
Fat content dropped in barley sprouts significantly than non- germinated grain; no change in fatty acid composition at lower temperatures and shorter times
Lipids Crude fats decreased by 5.73% compared to control on 1st day of germination
Higher levels of vitamin B1, B2 and C at longer germination times and higher temperatures
Vitamins and minerals –
Rise in TPC of barley (4.4–9.5 μmol GA Eq/g) after germination; increase in antioxidant potential in sprouted barley
Phytochemicals –
Rico et al. (2020)
References Waleed et al. (2021)
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Sprouting Sample conditions Black barley Soaking in ultra-pure water and (Hordeum distichum L) hydrogen rich water at 25 °C and geminated in dark
Changes in Proteins and amino acids Decrease in protein levels in both cases; increased the levels of nonessential, essential and semi-essential amino acids in ultra-pure water treated sample Carbohydrates and fibre Reduction in starch contents in both groups; hydrogen rich water treated sample showed more decrease in starch levels; reduced total and soluble dietary fibre in hydrogen rich water treated sample Lipids Reduction in lipid in both groups; hydrogen rich water treated sample showed more decrease in fat levels
Vitamins and minerals Noticeable reduction in riboflavin and thiamine in both groups; but on 6th day of germination, the contents of thiamin and riboflavin were significantly raised in both cases; substantial rise in Ca, Cu, Zn, Fe, and Mn levels in both cases
(continued)
Phytochemicals References Guan et al. Presence of (2019) isopimpinellin, methyl jasmonate in hydrogen rich water treated germinated barley, which possess antiviral properties; reduction of free chlorogenic acid and ferulic acid in ultra-pure water and hydrogen rich water treated samples; DPPH based antioxidant potential increased by 58.5% and 37.4% in hydroegn and ultra-pure water treated samples, respectively
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Hulled barley
Sample Barley (Hordeum vulgare L. ssp. distichum) and (Hordeum vulgare var. nudum)
Sprouting conditions 5% H2O2 sanitized seeds were soaked (6 h) in dark, After draining, some soaked seeds (40 g) were spread on filter paper and left overnight in darkness. Germination lasted for 7 days, including 6 days light and dark modes rhythmically alternated. Harvested sprouts were freeze-dried and ground. Soaking in distilled water for 16 h, then germinated for 1 day at 25 ± 2 °C and dried at 40 °C; some grains were treated with microwaves (950 W for 2 min)
Table 2.1 (continued)
–
Changes in Proteins and amino acids – Vitamins and minerals –
–
Carbohydrates and fibre Lipids – –
– β-D-glucan content was 3.48%, 2.85% and 3.87% in unprocessed, germinated and microwave treated barley
Ahmad et al. β-D-glucan from (2016) germinated barley showed significantly higher reduction potential than β-D-glucan from microwaved barley; metal chelation action of germinated barley was higher (31.34%) than microwave treated sample (23.65%)
Phytochemicals References Aborus et al. 1.48 times higher total phenolic content (TPC) in (2017) Hordeum vulgare L. ssp. distichum than in Hordeum vulgare var. nudum Both varieties contained vanillic and ferulic acids, catechins and gallic acid as dominant polyphenols.
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Barley (Grimmett)
Barley (Schooner)
Sample Barley (Hordeum vulgare L.)
Firstly steeping was done for 24 h followed by sprouting at 19 °C for different time intervals i.e. 0, 2, 4, 6 and 8 days Firstly steeping was done for 24 h followed by sprouting at 19 °C for different time intervals i.e. 0, 2, 4, 6 and 8 days
Starch decreased by 58.6% on 8th day of germination
–
–
Crude fats increased by 18.75% compared to control at 8th day
Vitamins and minerals –
Crude fats increased by 10% compared to control on 8th day of germination
Carbohydrates and fibre Lipids – –
Starch decreased Crude protein by 7.14% at 8th decreased by 9.43% at 8th day day
Crude protein decreased by 40.74% on 8th day of germination
Changes in Sprouting Proteins and conditions amino acids – After steeping grains in water for 1 h at room temperature, the germination performed under dark in a room for 0 (control), 12, 24, 36, 48, 60, and 72 h at 25 °C and 90% RH
–
Phytochemicals Firstly, rise in TPC with germination time for initial 48 h and then started to decline; all samples showed DPPH antioxidant potential but sample germinated for 48 h exhiibted significantly higher activity than others and non-germinated extract had the lowest antioxidant activity –
(continued)
Chu et al. (2014)
Chu et al. (2014)
References Ha et al. (2016)
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Barley (Hordeum vulgare L.)
Sample Barley
Sprouting conditions Pre-cleaning of barley grains was done using 1.25% sodium hypochlorite. After being cleaned, the grains were soaked in tap water for 24 h at 16.5 °C. The process of germination took place for 5 days in the dark, at 98% RH and 16.5 °C. The grains were manually aerated once every 24 h and sprayed with tap water for 15 min at 12-h intervals. Barley was soaked for 6, 12, and 24 h in distilled water at a ratio of 1:4 (w/v). The sample was germinated with water sprayed three times daily at 25, 30, and 35 °C.
Table 2.1 (continued)
Crude protein content was increased by 14.31% compared to control in 24 h soaking time at 25 °C
Changes in Proteins and amino acids Crude protein increased by 24.73%
Total carbohydrate content was increased by 8.90% compared to control in 12 h soaking time at 30 °C
Crude fat content was increased by 13.60% compared to control in 24 h soaking time at 25 °C
Carbohydrates and fibre Lipids Starch increased Crude fats by 60.18% increased by 27.00%
Crude ash content was increased by 15.34% compared to control in 24 h soaking time at 35 °C
Vitamins and minerals Ash content increased by 7.87%
References Donkor et al. (2012)
Singkhornart and Ryu (2011)
Phytochemicals –
–
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amorphous and crystalline portions of starch are solubilized by the α-amylase. The α(1–4) and α(1–6) linkages are broken by the α-glucosidase that is present in the embryo when it is acting on the non-reducing ends. The embryo contains β-amylase, which hydrolyzes the α(1–4) bonds to produce maltose (Gupta et al., 2010). Debranching enzymes work on the α(1–6) linkages in the amylopectin while α-amylase is found inside the aleurone and acts on the α(1–4) linkages. It is notable that neither the process nor the timing of the selective starch hydrolysis (crystalline or amorphous) during the malting are entirely known. During germination, the hydrolysis of starch does not fully convert it to maltose or glucose (Contreras- Jiménez et al., 2019). After germination, some starch granules remain intact, according to scanning electron microscopy, whereas others mostly the tiny starch grains are damaged by the enzymatic reaction. The variations in amount of soluble carbohydrates during the sprouting and establishment of barley seedlings were identified by Shaik et al. (2014). Due to a partial breakdown of starch into simple sugars during seed sprouting, they saw that these levels surged during the initial days of sprouting and suffered a significant reduction from 9 to 12 days. On the other hand, due to the starch being broken down by amylases during germination, Watchararparpaiboon et al. (2010) discovered an increase in the concentration of reducing sugar. However, when the temperature of the steeping process increased, the amount of reducing sugar in geminated barley dropped. However, one of the primary elements regulating germination is temperature, which may have played a significant part in the formation of sugars. The part of food that remains undigested after passing through the stomach and small intestine is referred to as the dietary fibre, also termed as roughage or bulk. Therefore, despite being vital to human health, these chemicals do not add to the nutritional level of food. Dietary fibres can be divided into two categories: soluble and insoluble. Insoluble dietary fibre (IDF), also known as resistant starch, aids in the transit of food through the digestive tract, increasing the size of the stool and preventing irregular stools or constipation. Soluble dietary fibre (SDF) lowers blood cholesterol and glucose levels. Teixeira et al. (2016) found that sprouting barley grains for 3 days at 15 °C didn’t significantly affect the total fibre content, although Koehler et al. (2007) exhibited the reduction within initial 2 days of wheat sprouting at 15–20 °C. Barley sprouting had a significant positive impact on cellulose levels, increasing it by 188% (Ortiz et al., 2021). The ingestion of starch and the increase in structural carbohydrates synthesis in sprouts are two potential causes of this. Barley was sprouted for 6 days by Ortiz et al. (2021), which shown 50% decrease in the amount of ß-D-glucan. A similar reduction of ß-D-glucan content was noted by Senhofa et al. (2016) when sprouting barley for 48 h. The sprouting time in the later trial lasted just 48 h, as opposed to the initial study’s 6 days, which may help to explain the disparity in results for the influence of sprouting on ß-D-glucan of barley. It appears that the mobilisation of cell-wall soluble polysaccharides and subsequent breakdown into low-molecular components for use as an energy source is the cause of drop in the ß-D-glucan levels of barley grains while sprouting (Guine & dos Reis Correia, 2013).
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2.5 Changes in Proteins Proteins are comprised of multiple building blocks – amino acids which attracts a great attention worldwide due to their nutritional importance. Based on the cultivar, barley contains 8–12% more protein than other cereals (Contreras-Jiménez et al., 2019). It is pretty apparent that germination reduces protein content because sprouting produces endopeptidases. These are crucial for the development of seedlings because they enable the functioning of stored proteins by breaking them down (Faltermaier et al., 2015). These endopeptidases have optimum activity at 40–50 °C and pH 3.5–6.5. Hordeins, which are predominately alcohol-soluble prolamins are found in barley and are high in glutamine, lysine, proline, and cysteine levels. By the fifth day of germination, half of all seed protein had been broken down. Following this time, the protein content of the seed rapidly declined from 3.5% on the fifth day of germination to 1.9% on the tenth day. The ratio of globulins and prolamins was consistently decreasing during seed germination, but the ratio of albumins and glutelins was trending in the opposite direction (Yogesh & Matta, 2011). Additionally, Ortiz et al. (2021) discovered that sprouted barley has 38.6% more protein compared to native grains. As a result, only glutamic acid dropped (P > 0.05) during sprouting, while the concentrations (g/kg, DM) of aspartic acid, valine, threonine, isoleucine, alanine, lysine, and tryptophan raised. But there are many studies showing upsurge in protein (5–10%) of sprouted barley (Donkor et al., 2012; Teixeira et al., 2016). The increase can be explained by the respiratory losses of carbohydrates, but the decrease in protein was caused by the leaching of water-soluble peptides in the steeped water (Afify et al., 2012). Proteins are more easily soluble and digestible due to sprouting. In this regard, barley and sorghum have both shown a 1.2- to 2-fold increase in protein solubility on sprouting the grains at 17–27 °C for 3–5 days (Osman et al., 2002; Afify et al., 2012).
2.6 Changes in Lipids Lipids are hydrophobic biomolecules which are composed of triglycerides as oils, waxes, phospholipids, sterols, etc. Lipids make up the 3rd greatest portion of barley after proteins and carbohydrates. Different barley varieties have a fat content ranging from 1.5% to 3% (Contreras-Jiménez et al., 2019). A regulated metabolic process is required for mobilising the triglycerides from oil bodies. This action is initiated with germination and results in the net conversion of oil to carbohydrates (Graham, 2008). The esterified fatty acids (FAs) from triglycerides are first released by lipases. The β-oxidation and glyoxylate cycles can then be used to break down free fatty acids (FFAs), which can then be turned into sugars (Graham, 2008). Barley germinated over 5 days at 22 °C experiences an 8–15% drop in the lipid levels as a result of the lipase activities (Chung et al., 1989) However, according to Watchararparpaiboon et al. (2010), crude fat levels in germinated barley did not
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vary considerably from those in ungerminated barley. According to Ortiz et al. (2021), the sprouting process also had an impact on the amount of fatty acids present in the raw barley grain. During the 6 days of sprouting, the contents of C18: 3 n-3 and C18: 0 rose by 49% and 24%, respectively, whereas the contents of C18: 1 n-9 fell by 6%. Sprouting had no effect on the amount of linoleic acid, which is the main fatty acid found in raw as well as sprouted forms of barley. Polyunsaturated fatty acids (56.1–61.4%) comprises the majority of morphological fragments (green shoot, root fractions, and residual structures) of sprouts. Saturated fatty acids and monounsaturated fatty acids were in less amount in the green shoot fraction. Besides this, it was also noted that the amount of fat increased during the sprouting of barley (Fazaeli et al., 2012) as a result of both an increase in the production of structural lipids linked to seedling development and compositional variations brought on by the breakdown of many other chemical components.
2.7 Changes in Minerals and Vitamins The mineral or ash content represents the total inorganic matter present in any substance. It is basically the naturally occurring bio-element, considered as an essential nutrient to properly regulate the metabolic mechanisms and physiological methods. Adequate intake of minerals is vital to maintain the cell protection, functionality, health and homeostasis whereas their deficiency is the root cause of several disorders. For example, cobalt (Co), copper (Cu), iron (Fe), manganese (Mn), and zinc (Zn) are essential for body growth and development, reproduction, and blood formation, while calcium (Ca), magnesium (Mg), and phosphorus (P) in combination, aid in the growth and maintenance of bones and regulating the osmotic pressure of blood, cellular and intercellular fluids (Ciosek et al., 2021; Jomova et al., 2022). However, the phytates highly bind the metal ions in grains like barley, which has a detrimental impact on the bioactivity of mineral elements including Ca2+, Cu2+, Fe2+, Mg2+, Zn2+, and Mn2+. Due to their strong propensity for cations, phytates are considered an antinutrient factor (Ikram et al., 2021). Phytase activity is eight times higher in barley during the first few days of sprouting than it is at the beginning of the process (Boukid et al., 2017), yet phytate content decreases throughout germination to a different level. As per Farooqui et al. (2018), the sprouting process raised the concentration of macroelements like Ca, Mg, and P from 110 to 130, 160 to 180, and 500 mg/100 g, respectively, but K decreased from 300 to 250 mg/100 g in germinated barley. In contrast to zinc, which rises to 3.48 from 2.92 mg/100 g, the microelements Fe, Mg, and Cu decreased with germination from 8.70 to 7.1, 1.54 to 1.49, and 0.68 to 0.62 mg/100 g, respectively. Crude ash, however, did not differ significantly from raw barley in another study on germinated barley (Watchararparpaiboon et al., 2010). Vitamins refer to the organic molecules, required in small amount for proper growth and development of body. These compounds have different biological functions and chemical composition which is important for synthesis of essential
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cofactors, enzyme and coenzymes derived metabolic processes (Zempleni et al., 2013). These exist in nature in their natural state or as pro-vitamins or precursors. Only the catalytic actions of vitamins are carried out; they help with the synthesis and destruction of substances, resulting in the body’s metabolism of fats, carbohydrates, and proteins. Adults meet 3–29% of the Recommended Dietary Allowances for vitamins while consuming 100 g of barley. The steeping and sprouting circumstances during sprouting determine the changes in vitamin levels. In cereals like barley, vitamin C levels are normally insignificant or very low (Ikram et al., 2021), however vitamin C is generated during sprouting. The higher ascorbic acid levels were found in barley sprout, according to Danisova et al. (1994), likely as a result of its de novo synthesis. This vitamin is highly heat-sensitive and light-sensitive vitamin, so controlled variables must be carefully selected to preserve it. The B-complex vitamins rise significantly as a result of sprouting, which boosts seedling growth and productivity. Barley’s riboflavin content increased after 4 days of sprouting at 17–28 °C (Hucker et al., 2012).
2.8 Changes in Phytochemicals The polyphenolic compounds found in barley, such as flavonoids and phenolic acids, are abundant. It was discovered that germination increased the antioxidant and polyphenolic concentrations of barley. After germination, barley was shown to contain the hydroxybenzoic and hydroxycinnamic acids, flavan-3-ols, flavanols, procyanidins, and flavones (Ge et al., 2021). The de novo synthesis and transformation may be responsible for the rise in these polyphenolic metabolites during germination (Gan et al., 2017). As a result of possible enzymatic cell wall disintegration while sprouting that released phenolic compounds and their level dramatically increased (Xu et al., 2009). Although few polyphenolic compounds were not present after sprouting, this may be because they underwent restricted oxidation and breakdown while sprouting was taking place (Gan et al., 2017). Guan et al. (2019) discovered that the in-vitro antioxidant potential of black barley were improved as well as increased quantities of several bioactive constituents like free coumaric, conjugated vanillic, vanillic, syringic, and sinapic acid. By using hydrogen-rich water, they improved the black barley’s capacity to germinate more successfully at low temperature of 10–15 °C. However, this significantly reduces the amount of dietary fibre and increases the amount of calcium and iron in black barley that has been sprouted. Sprouts, as found in in-vitro studies, have a larger phytochemical concentration and better anti-inflammatory benefits, according to Aborus et al. (2017). Rico et al. (2020) also found that barley flour has excellent antioxidant qualities and a low glycemic index (GI) when the germination duration is between 0.8 and 6 days and the temperature is between 12 and 20 °C. During pre- and post-germination, barley flour’s total phenolic content, antioxidant activity, and flavonoid concentration varied from 2.12 to 1.85 mg ferulic acid equivalents/g, 11.37–14.36%, and 0.29–0.37 mg catechin equivalents/g (Farooqui et al., 2018). Similarly, Sharma and Gujral (2010)
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reported 34.28% DPPH scavenging activity from 16.9% in barley when sprouted at 24 °C on 2nd day.
2.9 Changes in Other Components The process of germination promotes the biochemical processes that enhance some nutrients including ferulic, phytic, and particularly γ-aminobutyric acid (GABA) (Watchararparpaiboon et al., 2010; Singkhornart & Ryu, 2011). Glutamate decarboxylase – an enzyme that catalyzes the L-glutamic acid decarboxylation, produces GABA, a non-protein amino acid molecule. GABA appears to control anxiety, relaxation, the immunity, fat metabolism, increase the brain metabolism, prevent the headaches or the consequences of cerebral arteriosclerosis, and plays a significant part in neuronal function and activation of hypotensive and diuretic impacts (Ma et al., 2015; Ochoa-de la Paz et al., 2021). In addition, the activity of the enzyme phytase, which is involved in the breakdown of phytate, tends to rise during germination. Sung et al. (2005) observed in barley a relatively low initial phytase activity that grew to an eightfold level within the first few days after sprouting. As a result, varying amounts of phytate concentration are reduced during germination. Minerals and phosphorus are more bioavailable as phytate level is reduced. In barley, sprouting for 3–7 days at 15–27 °C has been shown to boost peptidase activity by a factor of 2–5 (Osman et al., 2002). After germination, the amylase activity of barley rose. When barley was soaked at 25 °C rather than 30 or 35 °C, the enzyme activity following germination was higher. It is commonly known that during germination, cereals produce enzymes whose activity is affected by environmental parameters such as temperature, moisture, the type of chemicals employed, genotype, and environment (Watchararparpaiboon et al., 2010). Amylases reduce the paste viscosity of barley, as well as maize and wheat sprouts, by breaking down the amylose and amylopectin of the starch to produce smaller dextrins, maltose, and glucose (Ma et al., 2020).
2.10 Techno-functionality of Barley Sprouts Barley undergoes biochemical changes during sprouting that alter not only its physical and chemical composition but also its nutritional value as well as utilization to develop different food products. The barley flour’s brightness (L*) was reduced by germination. However, as the germination progressed, the intensity of redness (a*) and yellowness (b*) of flour augmented (Rico et al., 2020). During germination, starch and protein hydrolysis is responsible for such consequences, which are then followed by the production of Maillard reaction chemicals after drying treatment. Additionally, bran colours migrate into the water and the endosperm during the steeping time.
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The process of sprouting shows an improvement in the protein’s solubility and digestibility. Barley has also shown 1.2- to 2 times augmentation in the solubility of proteins when grown at 17–27 °C for 3–5 days (Ikram et al., 2021). The regular barely flour absorbed water at a rate of 70.30%, while germinated barely flour did so at a rate of 54.80%. After germination treatment, water absorption capacity (WAC) thus decreased (Lotfy et al., 2021), and this drop may have been brought on by the breakdown of starch in sprouting seeds. According to this theory, Fincher and Stone (1986) and Amri et al. (2016) demonstrated that barley grains when sprouted, a group of hydrolytic enzymes (produced in aleurone layer and scultellum of germinating grain) mobilise the nutritional reserves of the endosperm, primarily starch and proteins. Yaqoob et al. (2017) used the various ratios of wheat flour, taking raw and sprouted barley flour. The pasting qualities of flour blends for making cookies were greatly improved by incorporating sprouted barley flour. Peak viscosity, a measure of how easily starch granules dissolve and which is frequently associated with the quality of the finished product, rose when raw (1224–1312 cP) and sprouted (1210–1258 cP) barley flour were used in comparison to control wheat flour (1212 cP). Because sprouting causes starch to break down due to enzyme activity, the peak viscosity of wheat flour and sprouted barley blends is a little lower than that of unsprouted barley and wheat flour blends. Additionally, the sprouting shortens the molecular chain length of β-glucan, which reduces the viscosity of sprouted barley flour than regular barley flour (Yaqoob et al., 2017). A different study found that β-glucan suspensions made from barley that had been sprouted for 24 h at 25 °C showed 40% lesser viscosity values compared to a solution made from untreated barley. Sprouting decreased the mean molecular weight (MW) of β-glucan and, thus, its potential to thicken by 22% (Ahmad et al., 2016). In another work, Park et al. (2022) created a 3D-printed cheese cake using powdered barley sprouts as the printing ink and examined the physico-chemical and functional aspects. Barley sprout powder’s functional characteristics, such as its water and oil holding capacity as well as water solubility index were 2.25%, 2.02% and 29.55%, respectively. Therefore, it can be said that sprouting affects the colour characteristics of barley in addition to enhancing its nutritional content. The solubility is increased during the sprouting phase, but the viscosity and water absorption properties are decreased. In order to give nourishment and regulate consistency in products like beverages, sprouted barleys are an excellent option.
2.11 Health Advantages of Barley Sprouting Germination is regarded as a realistic, economic and value-added method for enhancing the nutritional value and quality of barley. Sprouting contributes to enrich the some secondary metabolites, like as polyphenols and flavonoids, improving the grain’s antioxidant potential through the phenyl propanoid pathway and the acetate/ malonate pathway (Babenko et al., 2019; Glagoleva et al., 2022). At different stages
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of germination, these chemicals’ compositions and quantities significantly vary. The polyphenolic concentration of naked barley at different phases of germination varied substantially, according to Kruma et al. (2016). Germination produces useful secondary metabolites such phenolic acids, flavonoids, and condensed tannins. For the purpose of producing barley sprout ferment, Rico et al. (2020) optimised the germination time and temperature. The glycemic index of germinated barley flours was higher than that of the non-germinated samples. ORAC antioxidant capacity, total phenolics, and γ-aminobutyric acid increased by two to fourfold during sprouting. Bound ferulic acid concentrations increased whereas procyanidin B levels decreased during germination. The sprouting has increased the barley’s overall anthocyanin content. However, after germination, it was shown that naked barley had a substantially higher total antioxidant capacity (TAC), and hot air drying had a higher TAC content than infrared drying (Ge et al., 2021). It has been demonstrated that the phenolic bioactive compounds in barley sprouts can control chronic hyperglycemia (Ramakrishna et al., 2017). During germination, the plant embryo restarts growing after a period of quiescence, which is brought on by the ingestion of water. The substances with phytochemical qualities and health-maintaining benefits (glucosinolates and natural antioxidants) are discovered after germination. These substances can play an important role, among others, in the prevention of several diseases. Anti-nutritive substances like trypsin inhibitor, phytic acid, and tannins reduced in concentration during germination (Marton et al., 2010). Thus, germination can produce nutrients that are useful and good for the body, helping to maintain health. Sprouts’ nutrient concentration is very high even after they are eaten during the early stages of growth (Donkor et al., 2012; Pajak et al., 2014). Barley sprouts are rich in soluble fibre called beta-glucan, which helps control blood sugar levels and prevents the diabetes. Based on its effects on low-density-lipoprotein (LDL) and total cholesterol, soluble fibre reduces the incidence of CVD. Clinical studies have demonstrated that eating barley sprouts lowers the CVD risk and improves the lipid metabolism in persons with questionable cholesterol levels (Byun et al., 2015). Sprouts’ high concentration of hexacosanol and β-glucans improve blood cholesterol metabolism and encourage bile acid excretion. A cellular sensor of energy metabolism and a regulator of cholesterol metabolism, AMP-activated protein kinase, was shown to be affected by barley sprout extract, which contains polyphenols, by Lee et al. (2015). Mice’s intracellular total and free cholesterol concentrations were lowered by 19.65 mg/g of total polyphenols in barley sprout extract, which resulted in reductions of 24% and 18%, respectively, in intracellular quantities. Kanauchi et al. (2008) examined the anti-carcinogenic properties of germinated barley, a prebiotic based mixture that contains around 80% hemicellulose and insoluble protein fibre rich in glutamine. This food product made from germinated barley impacts the development of colon cancer in its early stages and hinders the transformation of hyper proliferative epithelia. In comparison to the control diet, its administration raised the content of luminal short chain fatty acids, particularly butyrate, which is formed in the colon, and encouraged the synthesis of acetate. The in-vivo research revealed decreased succinate synthesis as well as decreased expression of
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β-catenin, abnormal crypt foci, and β-catechin formations. Cecal B-glycosidase, heat shock protein 25 (HSP25) positive cells content, and activities of solute carrier and tumour suppressor gene slc5a8 all increased in the test group in comparison to the control. Additionally, sprouts made from two types of powder highland barley were the subject of a study by Aborus et al. (2017), which showed that sprouts (in vitro) have a higher phytochemical content and better anti-inflammatory benefits The presence of saponarin, a multifunction flavonoid present in barley sprouts further showed the anti-obesity activity in barley sprout hot water extract (BSE) when given to C57/BL/6N mice (Kim et al., 2021).
2.12 Challenges and Future Aspects Due to their numerous health advantages and desired taste, the sprouts are becoming more and more well-liked by people globally. Sprouts offer a number of benefits, including a better nutritional value, a fresh and succulent taste, quick development cycles, and easy growing environments. However, sprouts are significantly more susceptible to microbial growth, diminishing their microbial safety because the germination parameters are favourable for the bacterial growth. The microbial contamination of sprouts can come from a variety of pre- and post-harvest contaminant sources, such as handling risks, seeds, germination media, storage conditions, and soaking water. Before harvest, contamination can occur from water used for irrigation, soil, animal and human faeces, pests, and human intervention (Iwu & Okoh, 2019). On the other hand, the post-harvest contamination sources include transportation vehicles, ice, machineries, harvesting instruments and containers, water used to wash harvested commodities, and food handlers (Olaimat & Holley, 2012). The majority of sprout-driven outbreaks, as per FDA (Food and Drug Administration), were brought on by seeds that were contaminated with bacterial pathogens prior to germination (Miyahira & Antunes, 2021). Raw barley sprouts should not be consumed by those who are more susceptible, such as small children, the elderly, immune-compromised individuals, and those who are chronically ill. In addition to the sprouts’ microbiological quality, the pre-harvest sprouting results in severe financial losses since it reduces the grain production, end-use quality, and the seed viability for sowing (Nakamura, 2018). The temperature and humidity that are present throughout grain production, harvest, and storage or transit affect a seed’s ability to germinate. In order to maintain viability and vigour under unfavourable conditions, the premium seed is either dried to levels below a threshold or hermetically wrapped for storage and shipping. Temperature swings between the day and night, usually between 15 and 25 °C, coincide with seed maturation and filling, producing seeds with little or no dormancy. During physiological maturity and harvesting, the hot temperatures and moist situations may coexist, and barley may experience premature sprouting (Gualano & Benech-Arnold, 2009). The germination of grain grown under these circumstances can be acceptable, but it
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can later degrade quickly. Pre-harvest sprouting reduces sprouting energy, lowers extract levels, and produces malt of lower quality, all of which are undesirable in the manufacture of malted barley (Edney et al., 2013). The germination parameters must therefore be carefully managed for grain germination. Considering necessary precautionary actions like Good Agricultural Practices, Good Manufacturing Practices, Food Safety Management Practices on farm level, and Sanitation SOP (Standard Operating Procedures), post-harvest handling, as well as processing levels, are the first steps for preventing the incidences of bacterial contamination (EFSA, 2011). In order to lower the pathogen population and make seeds fit for human consumption, seeds need also be treated chemically, physically, or biologically before being used for germination. To enhance the quality of sprouts, new technologies must be developed or quickly adopted. A relatively recent effort to improve the nutrition and final quality of product involves the use of controlled germination process by treating grains with controllable physical energy. To encourage seed germination, these procedures apply energy in the forms of hydrostatic pressure, electrical energy, acoustic energy, cold plasma, light, etc. (Ding & Feng, 2019). Under ideal circumstances, cold plasma (CP) can encourage sprout growth, hence increasing the yield of sprouts. Similar to it, ultrasound processing did not affect sprouting and actually increased it. By enhancing water absorption and oxygen availability through sonic cavitation, ultrasound enhances the porosity of the seed. Cell membrane rupture aids in the mobilisation of endosperm nutrients. However, more research is necessary to ascertain the primary (physical and/or physiological) cause of the increased germination rate (Miano et al., 2015). CP can encourage the buildup of bioactive chemicals in sprouts, which will then improve biological processes and have an antiproliferative and antioxidant effect. Reactive species can efficiently be removed using CP by inactivating the microorganisms on seeds and sprouts (Liu et al., 2022). Barley seeds’ phenolic concentration and antioxidant activity can both be increased by germination (Ha et al., 2016). The identification of antioxidant compounds, however, was not entirely established yet. After soaking, germination, and kilning, relative to unprocessed samples, the amounts of individual tocopherols and total vitamin E contents were both decreased. However, once again, it is unknown what substances cause such variations in these vitamins (Do, 2016). Therefore, the usage of sprouts in various food formulations can be a suitable approach for improving their high nutritional value, provided their hygienic-sanitary safety is established. As an essential component of functional foods, the assurance of microbiological safety helps barley sprouts prevent chronic diseases.
2.13 Conclusion The use of barley sprouts or development of sprout-based formulations is expected to rise with respect to the consumer demand for natural, fresh and nutritious foods, free of pollution. Barley undergoes substantial improvements during germination,
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including the mobilisation of starch reserves by the action of α-amylase, a transition in the nitrogen-containing constituents toward oligopeptides and free amino acids, the hydrolysis of triacylglycerols resulting in increased saturated to unsaturated fatty acids ratio, a significant drop in anti-nutritional factors and an increase in minerals. Barley sprouts can therefore be utilised to make novel food products in addition to their traditional application in the production of barley malt, suggesting them as “promising functional foods.” But more investigation is essential to determine the optimisation of germination process and the pre- and post-harvest methodologies to lower microbial hazards without changing the nutraceutical compositions of sprouts. Additionally, there is very little evidence from clinical studies to support particular claims about the health benefits of barley sprouts, therefore researchers should focus in this regard. It is also important to comprehend how the recent processing techniques improve nutrition in germinated barley. Food scientists should study the functionality of barley sprouts during different food formulations to produce healthy foods.
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Xu, J. G., Tian, C. R., Hu, Q. P., Luo, J. Y., Wang, X. D., & Tian, X. D. (2009). Dynamic changes in phenolic compounds and antioxidant activity in oats (Avena nuda L.) during steeping and germination. Journal of Agricultural and Food Chemistry, 57(21), 10392–10398. Yaqoob, S., Baba, W., Masoodi, F. A., Bazaz, R., & Shafi, M. (2017). Effect of sprouting on barley flour and cookie quality of wheat–barley flour blends. Forum of Nutrition, 16, 175–183. Yogesh, K., & Matta, N. K. (2011). Changing protein profiles in developing and germinating barley seeds. Annals of Biological Research, 2(6), 318–329. Zempleni, J., Suttie, J. W., Gregory, J. F., III, & Stover, P. J. (Eds.). (2013). Handbook of vitamins. CRC Press.
Chapter 3
Buckwheat Sprouts D. Sowdhanya, Jyoti Singh, Prasad Rasane, Sawinder Kaur, Jaspreet Kaur, and Mukul Kumar
3.1 Introduction Germination is an ancient process practiced for softening the endosperm, increasing the bioavailability of seeds, and depleting the anti-nutritional compounds. The process initiates with the imbibition of water molecules in dry mature seeds, and swelling of grains due to water absorption and after a period of respiration, it activates endogenous enzymes like PAL and CHI leading to the sprout growth (radicle protrusion). Sprouts can be considered as “functional food” as it enhances the human health and body system and it possess some disease-preventing properties along with nutritional value owned by them normally (Miyahira et al., 2021). As quoted by Benincasa et al. (2019) some definitions for “sprouts” and “sprouted grains” were provided by western agencies and Associations. “Sprouts” (Regulation (EC) No 208/2013) are “the product obtained from the germination of seeds and their development in water or another medium, harvested before the development of true leaves and which is intended to be eaten whole, including the seed”. “Sprouted grains” are defined by the American Association of Cereal Chemists (AACC) with the endorsement of the United States Department of Agriculture (USDA) as follows: “malted or sprouted grains containing all of the original bran, germ, and endosperm shall be considered whole grains as long as sprout growth does not exceed kernel length and nutrient values have not diminished. These grains should be labelled as malted or sprouted whole grain”. As stated by Han and Yang (2015) seed germination is an essential parameter to determine the fertility of seeds and the proper environment should be provided in terms of water, temperature, light, soil, and nutrients to regulate the plant growth by influencing phytohormones such as gibberellic acids (GA) and abscisic acid (ABA). D. Sowdhanya · J. Singh (*) · P. Rasane · S. Kaur · J. Kaur · M. Kumar Department of Food Technology and Nutrition, Lovely Professional University, Phagwara, Punjab, India © The Author(s), under exclusive license to Springer Nature Switzerland AG 2023 I. Majid et al. (eds.), Advances in Plant Sprouts, https://doi.org/10.1007/978-3-031-40916-5_3
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To obtain good quality sprouts, some sprouting characteristics need to be considered as stated by Devi et al. (2015). Sprouting rate depends upon the age of the seeds, seed viability decreases during the ageing of seeds. The modification in the chemical composition will take place due to the activation of enzymes during germination. There will be elevation in protein, vitamin, and monosaccharides and depletion in the quantity of complex structure like starch, sucrose, and fat and anti-nutritional compounds. Chaplygina et al. (2020) explain that generation of active compounds which are energy and building material occurs due to the decomposition of macromolecules over a particular period. The monosaccharide elevation graph occurs after germination due to the enzymes present in scutellum and aleurone cells such as α-amylase and α-glucosidase activation in denovo synthesis. In addition, detachment of disulfide linkages holding β-amylase and protein together occurs. All the released enzyme acts on the respective compound, hydrolysis takes place resulting in higher production of simple sugars (Saithalavi et al., 2021). Buckwheat (Fagopyrum esculentum Moench.; 2n = 2x = 16), is an ancient pseudo crop belonging to the Polygonaceae family and Fagopyrum genus (Mizuno & Yasui, 2019). This dicotyledonous crop of both perennial and annual species found to have both diploid (2n = 2x = 16), and (tetraploid 2n = 4x = 32) chromosome numbers encountering an average haploid genome size of 1.2 Gb (Singh et al., 2020). Zhu (2016) reported that crops of Poaceae family such as rice (Oryza sativa) and wheat (Triticum aestivum) and Polygonaceae family of buckwheat (Fagopyrum esculentum) possess similar morphological structure and chemical composition with some difference. They both comprises of starchy endosperm, and non-starchy fiber rich aleurone layer. Buckwheat seeds are rich in high quality protein with well-balanced amino acids, starch composition, fiber, vitamins, minerals, and bioactive components. Species of outcrossing, self/cross pollinated traditional crop composed of bioactive components such as flavonoids, phytosterols, myoinositol, D-chiro- inositol, free and bound phenolic acids, and phenylpropanoid glycosides. The consumption is gradually increasing worldwide considering the health benefits of buckwheat and cultivated as major crop and functional food source (Li, 2019).
3.1.1 Buckwheat Sprouts Government of India declared “Nutri Cereals” as two pseudocereals namely amaranth and buckwheat. As buckwheat contains abundant nutritive value and it is feasible for the stable production and supply, hunger of poor community people will be quenched. The biological composition of every cereal contains few antinutritional compounds which reduces the bioavailability by binding together. Shreeja et al. (2021) states that most widely accepted method to deteriorate antinutritional compound and pumping up the functional compounds is possible by germination. Briatia et al. (2017) states that the ideology of introducing buckwheat sprouts as a new vegetable product was done by Kim et al. (2004). The physical appearance of
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sprout resembles the yellow colour with attractive fragrance, soft and crispy texture, devoid of bean flavour. It is considered as vegetable product due to the detachment of pericarp from the cotyledons. In Northeast Asia, these sprouts are consumed in the form of fresh vegetable or as salad along with noodles (Nam et al., 2015). It has been consumed as traditional food in Tsugaru District, in Aomori Prefecture, in the north of Japan to combat the shortage of vegetable crop during winter. It is germinated using hot spring heat and it also recommended to consume in higher quantity as it has possibilities in prevention of cancer as stated by Yamanouchi et al. (2018). Sprouting of buckwheat is practiced and recommended by many scientists due to the exponential rise in bioactive compounds due to several biochemical and enzymatic reactions. There is an increase in γ-aminobutyric acid (GABA), free amino acids and flavonoids especially rutin (Ma et al., 2020). There is an expansion in the accumulation of flavonoids as stated by Mansur et al. (2019) namely orientin, isooreintin, vitexin, isovitexin, Quercetin-3-O-robinobioside, and rutin which is directly proportional in increasing antioxidant potential with 5–7 days of germination. In addition, total phenolic content (TPC) contributes a way higher to antioxidant potential, in buckwheat sprouts significant amount of TPC was detected. The higher count of TPC is due to the rupture of cell wall releasing phenolic compounds (Chen et al., 2022). Explanations of Jang et al. (2019) reveals that flavone C-glucosides and phenolic compounds promote buckwheat sprouts as therapeutic food possessing antioxidant potential and anti-inflammatory activity. Considering these health beneficial components, Nam et al. (2018a, b) quotes that buckwheat as “popular health food”.
3.1.2 History and Origin The Swiss plant taxonomist De Candolle claimed that the buckwheat was originated from Siberia and northern part of China. In 1957, Nakao opposed his statement and expressed that lot of wild buckwheat species were widely spread in southern part of China and later it was considered as the place of origin for buckwheat crop (Zhou et al., 2018). The transitional agricultural growth in China started from the cultivation of dry millets in Loess plateau and Yellow river catchment in North China and rice cultivation in middle and lower Yangtze valley followed by growth of roots and tubers in third centre of southern China along the Zhujiang river south of Nanling mountains (Hunt et al., 2018). The birthplace of buckwheat was assumed to be native to temperate east Asia in particular eastern site of Himalayas and Southern China. The conclusion on domestication of buckwheat was determined based on the discoveries on wild species of common buckwheat. The cultivated species of common buckwheat was found to be originated in Yunnan province on the edge of Tibetan plateau and Sichuan province of southwestern China (Suvorova & Zhou, 2018). The origin of Tartary buckwheat was found to be in Daliangshan region, borderlands of Yunnan and Sichuan province based on the growth and cultivation of large amount of buckwheat species. The cultivation of this species was performed
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since 5000–6000 years ago. According to the present data, 10,000 accessions were identified and maintained globally. Out of that 2800 accessions are being preserved by Institute of Crop Sciences, Chinese Academy of Agricultural Sciences (CAAS) (Dar et al., 2021).
3.1.3 Geographical Distribution The cultivation and development of buckwheat crop was diffused to various countries and continents through different routes. It was diffused eastward to Korean and Japan, westward to Europe and North America, southward to Indian subcontinent (Semjon, 2021). Suvorova and Zhou (2018) described the journey of buckwheat from China, it started diffusing from China to Korean peninsula where it is widely called as hunger crop. Further, it travelled into Japan and spread to western part of Central Asia. The route way continued from Central Asia to Tibet, Bhutan and entered Nepal and India. There is historical evidence of cultivation in Japan since early Jomon period (7000–5000 years BP) (Woo et al., 2016). The crop moved to Europe through Siberia and south of Russia around 1200–1300 AD. From Europe, it started entering countries like Ukraine, Germany, Slovenia, followed by Belgium, France, Italy, United States and Dutch in seventeenth century (Zhou et al., 2018). The growth of wide varieties of buckwheat species can be noticed in the mountainous States of North-Western regions, viz. Jammu and Kashmir, Himachal Pradesh, Ladakh and Uttarakhand. The scattered growth in the regions of North Eastern states of Sikkim, Assam, Arunachal Pradesh, Nagaland, West Bengal and Manipur and intermittent cultivation in Chhattisgarh and Tamilnadu (Dar et al., 2021).
3.1.4 Taxonomy and Botanical Classification Buckwheat belongs to the tribe of Polygonaceae, family of flowering plants and subfamily of Polygonoideae. The term genus Polygonum was mentioned in the book Genera Plantarum by Antonie Laurent de Jussieu in 1789 (Semjon, 2021). The subfamilies of Polygonaceae family was classified into two divisions based on their morphological characteristics namely Polygonoideae Eaton and Eriogonoideae Arn (Wang et al., 2017). The Polygonaceae family has seven genera namely Antenoron, Fagopyrum, Fallopia, Koenigia, Polygonum, Pteroxygonum, and Reynoutria. In early days there were several generic names provided by different scientist namely Fagopyrum Tourn by Tourn (1742), Polygonum Linn by Linn (1753), Fagopyrum Miller by Miller (1754) and Fagopyrum Moench by Moench (1756). Zhou et al. (2018) reported that there were oscillations in determining the genera name and in considering as an individual species or grouping under Polygonum species. After the broad research, scientist Yukio find out the difference in basic chromosome number of Polygonum and Fagopyrum. Polygonum has chromosome number of
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n = 10, 11, 12 whereas Fagopyrum has n = 8 chromosome number. Based on the morphological, palynological, and cytological studies Fagopyrum is considered as an individual species. The wide range of species of this genus broadly categorized into two phylogenetic groups, the cymosum group and urophyllum group (Semjon, 2021) as mentioned in the Table 3.1. The cultivated species of agriculturally significant buckwheat are common buckwheat or sweet buckwheat (F. esculentum Moench) and Tartary buckwheat or bitter buckwheat (F. tataricum) (Zhu, 2016). The synonym for sweet buckwheat are F. esculentum subsp. Ancestralis Ohnishi and for bitter buckwheat are Polygonum tataricum L., F. suffruticosum, F. Schmidt, F. dentatum Moench, Fagopyrum rotundatum Bab., and F. subdentatum Gilib (Luitel et al., 2017).
3.1.5 Common Names Used in Different Regions of the World Buckwheat is a ubiquitous crop grown across worldwide due to the presence of several nutritional, functional and nutraceutical properties and it was named in different ways in every region based on its geographical condition and so on (Table 3.2).
3.1.6 Morphological Characteristics Buckwheat is an annual herbaceous plant with free branches growing from 0.5 to 1.5 cm tall. The green to red coloured stem with sucker for adaptation to thrive in unfavourable conditions have a stem diameter between 0.3 and 0.5 cm. Leaf is a simple, petiolate, green coloured and heart shaped leaf with a length of 2–8 cm long Table 3.1 Fagopyrum species Group name Cymosum group Urophyllum group
Species name F. esculentum and F. tartaricum F. cymosum, F. homotropicum, F. lineare, F. pilus, F. acutatum, F. callianthum, F. capillatum, F. caudatum, F. crispatifolium, F. densovillosum, F. dibotrys, F. emarginatum, F. giganteum, F. gilesii, F. gracilipedoides, F. gracillipes, F. hailuogouense, F. hybridum, F. homotropicum, F. jinshaense, F. leptopodium, F. lineare, F. luojishanense, F. macrocarpum, F. megaspartanium, F. pleioremosum, F. pugense, F. qiangcai, F. rubifolium, F. static, F. sagittatum, F. urophyllum, F. wenchuanense, F. zuogongense
Species category References Cultivated species Wild Luitel et al. species (2017) and Semjon (2021)
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Table 3.2 Common names of Fagopyrum esculentum (buckwheat) Country United States
Name Sweet buckwheat or common buckwheat, Tartary buckwheat or bitter buckwheat China Tian qiao (common buckwheat), kuqiao (tartary buckwheat) Nepal Mithe Phaper (common buckwheat), Tite Phaper (tartary buckwheat) India Ogal (common buckwheat), Phaper (tartary buckwheat) Bhutan Jare (common buckwheat), Bjo (tartary buckwheat) Japan Soba Russia Grecichakul’turnaja Ukraine Grechka Poland Tatarkagryka, gryka, poganka Czech Republic and Pohanka Slovakia Sweden Bovete Denmark Boghvede Finland Tattari (common buckwheat) Slovenia Ajda, hajdina, idina Bosnia, Serbia, Heljda Montenegro, Croatia France Sarrasin, ble’noir, renouee’, bouquette Berton (North West Gwinizh-du France) Italy Fagopiro, granosaraceno, Sarasin, faggina Germany Buchweizen or Heidekorn Korea Maemil
References Semjon (2021), Luitel et al. (2017) and Zhou et al. (2018)
and 3.4–5.2 cm wide (Luitel et al., 2017). The colour of leaf margin ranging from green, pink, and red colour. The shape of leaf blade is hastate and sagittate, leaf base is cordate or hastate (Dar et al., 2021). It is a heteromorphic self-incompatible species (SI) with an average number of inflorescences of 4–10. The incomplete flowers of Fagopyrum esculentum have colour ranging from white to pink. The flowers are positioned in the racemes at the ends of the branches or on short pedicles in a cluster formation. It has no petals but composed of calyx carrying five petals like sepal which is pink to dark pink in colour (Woo et al., 2016). The dimorphic species have floral architecture of two types namely thrum and pin. The characteristic of thrum and pin flower has short style, high anthers, short pistils, and large pollen grains and long style, low anthers, long pistils, and small pollen grains respectively (Takeshima et al., 2019). Buckwheat bears a triquetrous flower in acute angle called as achene which is 4–9 cm long. Achene is composed of two major parts such as hull and groat. Fruit coat and pericarp make up the portion of hull and the parts of seed coat, endosperm and embryo comes under the dehulled achene, groat (Tömösközi &
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Langó, 2017). Dar et al. (2021) findings reveal that the seed coat is 1–3 cm cell thick, and seed has the physical shape of triangular, conical and ovulate with grey to brown in color. The endosperm containing starchy rich granule seed has 1000 seed weight ranging from 18 to 26 g.
3.1.7 Production and Consumption The global leading producers of buckwheat are China, the Russia Federation, Ukraine, and Kazakhstan. The other major producers include Slovenia, Poland, Hungary, and Brazil. China is leading the chart globally for about 40 years in the production of buckwheat species (Semjon, 2021). Zhang and Xu (2017) reported as China is the worldwide largest producer in general and the higher concentration of production is found in the regions of Inner Mongolia, Shanxi and Shaanxi provinces. The major producers of buckwheat are China, Brazil, France, the United States, and Canada, and the largest quantity of 1,20,000 tonnes of buckwheat is imported by Japan per year (Woo et al., 2016; Suvorova & Zhou, 2018). In 2013, the leading producers were Russia, China, Ukraine and Poland with global production of 2,347,558 tonnes (Zhu, 2016). Tömösközi and Langó (2017) confess that crop production exceeded in 2014 by 2 million tonnes per annum with a cultivation area of 2.5 million ha (FAOSTAT, 2014). Suvorova and Zhou (2018) represented the statistical data of global production of buckwheat species in 2014. Ukraine contributed 2,00,000 tons of buckwheat globally, 30 thousand tons in Belarus. Poland with the annual production of 1,00,000 tons and it is dominating the crop production in European Union. In Japan buckwheat production was around 33 thousand tonnes, 10,000 tonnes production in Nepal, 4000 tons in Bhutan, 2000 tons in Korea, 1000 tons in Slovenia. Singh et al. (2020) expressed that France was leading producer in 2018 with an annual production of 3735 kg/ha. The figured estimation of India buckwheat production is not available. The cultivation of crop is dispersed from Jammu and Kashmir in the north to Arunchal Pradesh in west, and Tamilnadu in the south. In India, the distribution of crop is observed in the regions of Jammu Kashmir, Himachal Pradesh, Uttarakhand, West Bengal (Kalimpong, Coochbehar, New Jalpaiguri and Darjeeling region), Sikkim, Assam (Upper Assam), Arunachal Pradesh, Nagaland, Meghalaya (Higher elevation region), Manipur, Kerala, Tamil Nadu (Nilgiris and Palani hills) and Chhattisgarh (Babu et al., 2018). The top 10 leading producers in 2016 are Russia, China, Ukraine, Kazakhstan, Poland, the United States, Japan, Brazil, Lithuania, and France. The highest concentration of production was seen in Russia with 1.19 million tonnes production in 1.12 Mha followed by China (404,259 tons) and Ukraine (176,430 tons) (FAOSTAT, 2016). According to current data, the top 10 global leading producers are Russia (892.16 MT), followed by China (503.99 MT), Ukraine (97.64 MT), USA (86.40 MT), Brazil (65.12 MT), Japan (44.80 MT), Kazakhstan (40.09 MT), Belarus (28.30 MT), Tanzania Rep (25.77 MT) and Nepal (11.72 MT) (FAOSTAT, 2020) as represented in Fig. 3.1.
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Production status in 2020 (MT)
Russia
China
Ukraine
USA
Brazil
Japan
Kazakhstan
Belarus
Tanzania Rep Nepal
Fig. 3.1 Production Status of buckwheat in 2020 (in MT). (Source by FAOSTAT, 2020)
The underutilised buckwheat is consumed largely by Asian and European population in the form of groats, flour, noodles, and tea (Sim et al., 2020). The buckwheat consumption has gained popularity in recent days because of their qualitative nutritional contents, including high unsaturated fatty acids, amino acids, peptides, flavonoids, and other phenolic compounds (Qin et al., 2017) and it has high adaptive growing conditions such as climatic variables, infertile soil, fluctuating temperature, and water stress regime (Luitel et al., 2017). Starowicz et al. (2018) reported the commercial buckwheat products available in the market are bread, cake, gluten free bread, honey, tea, noodles, vinegar, sprouts, tarhana, alcoholic beverages such as beer and shochu.
3.2 Various Germination Treatments Involved in Sprouting Yiming et al. (2015) performed germination by treating the seeds through a sequential procedure of cleaning, soaking, and washing. The washed seeds were spread in flat containing moist paper towel, and it was covered with aluminum foil. The desirable conditions of temperature at 37 °C, and dark environment was given for 7 days. The experimentation of Yang et al. (2021) have shown that seeds were initially sterilized with 1% hypochlorite solutionfor 30 min. The sterilization was done to prevent the microbial contamination of bacteria, yeast, and fungi and to improve the seed germination (Jafari et al., 2016). With the aid of distilled water, soaking was done for 5 h at 28 °C under darkness. To obtain a proper sprout growth, germination
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was carried out at 25 °C and 28 °C, 80% relative humidity under 12 h cycle in dark environment. The equipped germination method was carried out by Chen et al. (2022), the preliminary step of soaking performed by immersing mature seeds in deionised water for 4 h under normal room temperature. The device set up containing incubator with water spraying device and water draining basket is used. The immersed seeds are spread evenly over the basket, to prevent the dryness and to improve germination rate water spraying device is fixed. The water is sprayed at frequent intervals and temperature is maintained at 25 °C. Almuhayawi et al. (2021) followed the germination method of laser irradiation which involves exposing the soaked seeds to laser beam. The light source of He-Ne was used to improve the crop production which is propagated in a longitudinal direction 12 cm away from the treated seeds. The pre-treated seeds placed in controlled growth chamber were maintained under some specified conditions such as temperature of 25 °C, relative humidity of 60%, photoperiod of 16 h light/8 h dark by white florescent tubes with photosynthetically active radiation till the sprout growth occurs. The experimentation with the aim of increasing anthocyanin content was performed by Seo et al. (2015) by dipping pre-treated seeds in L-Phe solution along with exposure to laser light of white, blue, red and red blue. The pre-treatment was carried out by soaking the seeds in sterile solution of 10% sodium hypochlorite at 25 °C for 3 h and optimum conditions in controlled growth chamber at 25 °C, 70% relative humidity under dark condition with a photo period of 18 h day/6 h night maintained for the sprout growth. The alternative method of soaking with 0.2% sodium bicarbonate (NaHCO3) for 5 h was carried out by Qin et al. (2017) and germinated the seeds by sowing in plastic boxes and covered with aluminium foil providing dark condition at 25 °C for 120 h and frequently spraying 0.2% NaHCO3 solution. Sterilized seed with 5% sodium hypochlorite for 30 min followed by immersing seeds in metallic additive solutions of Al2 (SO4)3, CuSO4 and ZnSO4 at 25 °C for 8 h to create stress conditions (Wang et al., 2013). Sathasivam et al. (2021) experimented with the addition of growth hormones such as 6-benzyl amino purine (BAP), kinetin, zeatin, and thidiazuron (TDZ) were treated to germinated seeds maintained in plant growth chamber at 25 °C in white cool fluorescent light of 35 μmol s−1 m−2 under photoperiod of 18 h light and 6 h dark followed by harvesting after 10 days of germination. The elicitors like salicylic acid, jasmonic acid and chitosan were used for treating buckwheat seeds by Park et al. (2019) followed by germinating in incubator at 25 °C for 72 h. The surface sterilization of seeds done with 10% sodium hypochlorite for 30 min and imbibed with water at 25 °C overnight. The swollen seeds were germinated at 25 ± 2 °C and 60 ± 2% relative humidity in plant pots for the period of 8 days. At every interval of 6 h, the combination of sucrose and calcium chloride solution should be sprayed for 5 min over the seeds in the replacement of water. Hao et al. (2016) performed germination on dehusked buckwheat seeds using slightly acidic electrolysed water by washing, soaking for 12 h, germinating at 20 ± 1 °C and 85–90% relative humidity and spraying with treated solutions once in a day.
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3.3 Chemical Composition of Buckwheat and Its Sprouts 3.3.1 Nutritional Composition of Buckwheat Buckwheat is composed of higher concentration of protein, carbohydrates, total and reducing sugars, dietary fiber, lipids, micro and macro elements, vitamins like vitamin B1, B2, B6, E, and C (Vollmannová et al., 2021). This pseudo crop possesses health benefit providing compound such polyphenol, flavonoid such as orientin, quercetin, vitexin, isovitexin, and isoorientin and anthocyanidins (Kumari & Chaudhary, 2020). 3.3.1.1 Carbohydrate The total carbohydrate present in the buckwheat grain and buckwheat sprouts ranges from 71–76% to 69.49%. The findings of Shreeja et al. (2021) reveals that there is a reduction in carbohydrate content of 4% after germination. The reduction is observed due to carbohydrate was utilized as a fuel source for the embryonic growth of sprouts. This crop has high calorific and carbohydrate content compared to staple food such as wheat and it can be replaced in the place of traditionally consumed wheat (Nepali et al., 2019). 3.3.1.2 Protein The second major component protein in buckwheat is soluble in nature with balanced amino acid profile. The proportion of amino acid is similar or comparatively higher than that of wheat containing essential amino acids such as lysine, threonine, and valine (Tömösközi & Langó, 2017). The raw buckwheat and buckwheat sprouts with 10–15% and 12–14% protein respectively contains major four protein fractions namely albumin, globulin, prolamin and glutelin. Podolska et al. (2021) reported the concentration of protein fractions with highest percent of albumin with 5.68–7.58% followed by glutelin with 1.32–1.67%, prolamin with 0.45–0.81% and globulin with 0.29–0.84%. The absence of gluten promotes the buckwheat as gluten free food for treating prophylaxis of gastrointestinal diseases in particular celiac disease. Sytar et al. (2018) claims that protein bioavailability of buckwheat is on higher scale rather than cereals with highest amino acid score (AAS) of 100. Mattila et al. (2018a, b) states that amount of protein can be affected by certain factors such as type of cultivar, grade of processing, environmental conditions. The findings of Shreeja et al. (2021) exhibits that there is positive increase in protein content after germination. The yield of high protein content can be achieved by optimising higher sprouting temperature, longer sprouting time and inhibition of water before germination. The physiological properties of protein found to be similar with dietary fiber. They have therapeutic properties such as hypocholesteromic,
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antiobesity, anticonstipation, and chemoprotective activity against tumorigenesis. The observations of Thakur et al. (2021) reveals that series of consequential biochemical reaction increases the protein content. It initiates with α-amylase activity on starch-protein complex followed by protease activity on cleavage of peptide bonds. Yiming et al. (2015) studies show that the germinated buckwheat has increased nutritional value with some decreased functional activity. Two significant reasons are considered for the increment in protein and amino acid content by Jin et al. (2022) which are degradation of polyphenol-protein complex results in reducing phytic acid and higher activity of protease on globulin and glutelin leads to push up the concentrations of soluble protein. 3.3.1.3 Fat The result of several findings estimates the lipid content of buckwheat was around 2.1–3.45%. In germinated buckwheat the lipid content was observed in the range of 2.05–3.09%. Shreeja et al. (2021) interpretations display that there is decrease in lipid content after germination. The depletion is due to lipid is deteriorated into CO2 and H2O during catabolic activity of sprouting. Omösközi et al. reviewed that lipid is concentrated in the embryonic region of seed in the bound form which is double the amount greater than free lipid content. Lipid breakdown due to lipolytic activity was consumed as energy source for the germination of seeds, this is results in the reduction of fat proportion (Thakur et al., 2021). Jang et al. (2019) explains that energy required for germination is produced by hydrolysis of triglycerides through tricarboxylic acid (TCA). 3.3.1.4 Fiber Buckwheat is a good source of dietary fiber with an average fiber content of 10/100 g (Nepali et al., 2019). Tömösközi and Langó (2017) stated that buckwheat has higher fiber content compared to amaranth and quinoa, lesser than whole wheat grain and like cereal grains. The fiber is present in free form without bounding to antinutrient compounds like phytic acid. The dietary fiber is found in larger amount in common buckwheat than Tartary buckwheat. The Tartary buckwheat has 8.4% dietary fiber composing of 8.2% insoluble dietary fiber and 0.2% soluble fiber (Li, 2019). The health beneficial dietary fiber comes under the category of resistant starch has potential effect in treating medical conditions like obesity and reduces the risk of colon colorectal adenoma and colorectal cancer (Zhang et al., 2020). The research findings of Kasar et al. (2021) shows that the concentrations of insoluble dietary fiber (ISDF) and soluble dietary fiber (SDF) increases in ascending order from endosperm, grit, bran, and hull. Fiber is highly concentrated in hull portion of buckwheat which is composed of complex cell wall material like cellulose, hemicellulose, lignin, and pectin. During germination there is a modification in these complex matrixes, interruption in protein-carbohydrate interaction leads to
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enhanced quantification of cellular structures like lignin, cellulose, and hemicellulose (Thakur et al., 2021). The germinated buckwheat sprouts reported with higher fiber content due to breakdown of complex starch molecules. The generation of new primary cells during germination results in the elevation of fiber content (Yang et al., 2021). 3.3.1.5 Energy The energy value is determined for raw and germinated buckwheat, and it was observed around 328–359.64 Kcal and 345.06 Kcal respectively. Shreeja et al. (2021) explained that after the germination the percent of total carbohydrate, energy, ash, and fat got diminished and the percent of protein, fiber and moisture got elevated. The moisture level gets increased after germination due to water uptake by the grain to carry out the metabolic activity (Table 3.3). 3.3.1.6 Starch and Sugars Buckwheat contains higher concentrations of storage carbohydrates which is accumulated in the endosperm region which acts as a storage reservoir and fuel energy for the plant growth (Semjon, 2021). The morphological structure of starch granules Table 3.3 Major nutrients present in buckwheat and its sprouts Buckwheat Buckwheat grains sprouts References 71–76 59.19–69.49 Sinkovič et al. (2021), Shreeja et al. (2021), Nepali et al. (2019), Nalinkumar and Singh (2020) and Verma et al. (2020) 10–15 12–17.46 Sinkovič et al. (2021), Sturza et al. (2020), Nalinkumar and Singh (2020), Shreeja et al. (2021), Nepali et al. (2019), Deng et al. (2015), Zhu (2016), Verma et al. (2020) and Thakur et al. (2021) Fat (%) 1.40–3.45 1.15–3.09 Zhu (2016), Sinkovič et al. (2021), Sturza et al. (2020), Shreeja et al. (2021), Nalinkumar and Singh (2020), Nepali et al. (2019), Deng et al. (2015), Verma et al. (2020) and Thakur et al. (2021) Energy (Kcal) 328–359.64 345.06 Shreeja et al. (2021), Nalinkumar and Singh (2020) and Verma et al. (2020) Fiber (%) 0.92–19.8 1.44–12.92 Sinkovič et al. (2021), Sturza et al. (2020), Nalinkumar and Singh (2020), Shreeja et al. (2021), Verma et al. (2020) and Thakur et al. (2021) Moisture (%) 9.24–11.03 10.18–12.77 Sturza et al. (2020) and Shreeja et al. (2021) Ash (%) 1.6–2.5 1.00–1.76 Sinkovič et al. (2021), Thakur et al. (2021), Sturza et al. (2020), Shreeja et al. (2021), Deng et al. (2015), Verma et al. (2020) and Thakur et al. (2021) Nutrients Total carbohydrate (%) Protein (%)
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varies from round, oval or polygonal with size ranges from 2 to 15 μm (Zhu, 2016). Nalinkumar and Singh (2020) describes the composition of starch molecules, with 15–25% of amylose, which is a predominant polysaccharide, 7–35% resistant starch (RS) and rest of the portion is amylopectin. An A type polymorph starch granules containing amylose with 12–45 glucose subunit in its chain and amylopectin with extra-long chain (DP > 100). Verma et al. (2020) recommended that consumption of 30 g of buckwheat in the daily diet as it is composed of retrograded and resistant starch. These kind of starch helps in the reduction of serum and LDL cholesterol, triglycerides and increases the level of HDL cholesterol which helps in reducing the risk of cardiovascular diseases. The combination of buckwheat protein and resistant starch have positive impact on diseases like hyperlipidaemia. Apart from starch, buckwheat is also enriched with sugar compounds which includes monosaccharides such as glucose, fructose, maltose, disaccharides such as sucrose, fagopyritol, mono, di, and tri-galactosyl derivatives of D-chiro-inositol. The fagopyritol content ranges from 128.2 to 277.4 mg/100 g (Raguindin et al., 2021). The galactosyl derivatives of D-chiro-inositol lowers blood pressure, increase glucose concentration, improves insulin resistance by enhancing the action of insulin, and increases the level of plasma triglycerides (Li, 2019). The findings of Kasar et al. (2021) reveal that the hydrophilic nature of polysaccharide contribute to good water holding capacity and the polysaccharides along with polar amino acid residues of protein contribute to water binding capacity (91.1%) due to their greater affinity to water molecules. Protein and carbohydrates can entrap the oil molecule by capillary action and show good oil absorption capacity (92.5%). Shreeja et al. (2021) stated that starch breakdown was performed by the catalytic activity of enzymes such as α-amylase and β-amylase which result in starch degradation after the germination process. Yiming et al. (2015) findings inferred that complex molecule breakdown into simpler form such as monosaccharides during germination. These simple sugar acts as sprouting energy during germination and gets accumulated in the buckwheat which result in the increased level of sugars after germination. Thou the higher sugar level is observed at the end of the germination due to starch hydrolysis, there is degradation of soluble sugars at initial stage due to anoxia (anaerobic metabolism), the stage between imbibition and rupture of hilum. The highest content of trehalose was found in the sprouts. The reduction in starch and amylose content was observed in the sprouts which is due to the energy supply required for germination. Comparatively amylose content has reduced in larger quantity than amylopectin, this exhibits that linear/slightly branched structure and smaller molecular size of amylose favoured the metabolic breakdown process (Yang et al., 2021) (Table 3.4). 3.3.1.7 Minerals The minor elements such as minerals are important to maintain physiological function in the human body. It is present in abundant in nature including several mineral elements such as Mg, P, K, S, Ca, Na, V, Fe, Co, Mo, Cr, Se, Cu, Mn, and Zn (Li,
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Table 3.4 Starch and Sugars present in raw and germinated buckwheat grains Buckwheat Nutritional composition Buckwheat grains sprouts References Starch (%) 42.83–69.77 39.25 Shreeja et al. (2021) and Deng et al. (2015) Total sugar (%) 1.84 3.24 Shreeja et al. (2021) Reducing sugar (%) 0.62 0.91 Shreeja et al. (2021) Glucose (%) 0.03–0.13 0.94–2.61 Sturza et al. (2020) and Mattila et al. (2018a, b) Fructose (%) 0.20 0.20–0.63 Sturza et al. (2020) Maltose (%) 0.11 0.05–0.06 Sturza et al. (2020) Non reducing sugar (%) 0.74 0.21–0.23 Sturza et al. (2020)
2019). Tomoskozi et al. stated the mineral comparison between amaranth and quinoa. The mineral composition is found to be higher than quinoa and similar to amaranth. The mineral deposition is higher towards the regions of bran than endosperm. The composition of mineral in plant seeds are affected by cultivating varieties and planting regions. Semjon (2021) expressed that foliar fertilization have shown increased level of Selenium (Se) and it shows beneficial effect in preparing nutrient enriched food product. Se enriched food have positive correlation in reducing chance of occurrence of heart diseases and hypothyroidism (Zhu, 2016). Nepali et al. (2019) described the nutritional characteristics of magnesium which could prevent arteriosclerosis and myocardial infarction, relaxes blood vessels, reduce the occurrence of headache, treat hypertension and depression. Copper aids in the production of red blood cells. The composition of mineral in buckwheat seed is mentioned in the Table 3.5. The treatments like soaking and germination have a positive impact on improving the mineral content. Thakur et al. (2021) studies prove that the trace minerals like copper, iron, zinc, and manganese have inflated to 153.77%, 97.67%, 61.42%, and 34.72% respectively. The bioaccessibility of iron from tartary buckwheat sprouts was found to be intermediate compared to its grain and groats as the Fe bio accessibility was higher in husk portion (Pongrac et al., 2016). 3.3.1.8 Vitamins Buckwheat is recognized as a good source of both fat soluble and water-soluble vitamins such as vitamin A, E and B complex, vitamin C respectively (Nalinkumar & Singh, 2020). Li (2019) discussed the occurrence form of vitamin E, lipid soluble antioxidant namely α-tocopherol, γ-tocopherol, and δ-tocopherol. Vitamin E has an antioxidant activity to fight against proliferative cells responsible for the cause of degenerative diseases such as cancer, tumor, oxidative stress, Type 2 diabetes, hepatic disease, and ageing (Zhang et al., 2020). Vitamin C also known as ascorbic acid, a natural antioxidant, its deficiency causing scurvy can be prevented by the
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Table 3.5 Mineral composition of buckwheat grains Nutritional composition Major minerals Magnesium (Mg)
Quantity (mg/g)
References
1.2–2.4
Phosphorus (P)
3.2–4
Sulphur (S) Potassium (K)
1.5–1.7 4.2–6.22
Calcium (Ca)
0.6–14.13
Trace minerals Sodium (Na)
Quantity (mg/kg) 21–36
Sinkovič et al. (2021), Podolska et al. (2021) and Verma et al. (2020) Sinkovič et al. (2021), Podolska et al. (2021), Zhang and Xu (2017) and Verma et al. (2020) Sinkovič et al. (2021) Sinkovič et al. (2021), Shreeja et al. (2021), Podolska et al. (2021) and Verma et al. (2020) Sinkovič et al. (2021), Podolska et al. (2021), Zhang and Xu (2017) and Verma et al. (2020) References
Vanadium (V) Chromium (Cr) Manganese (Mn)
0.05–0.09 0.20–0.40 12.22–18.7
Iron (Fe)
22–86.9
Cobalt (Co) Copper (Cu)
0.04–0.05 0.5–8.7
Zinc (Zn)
0.2–63.07
Molybdenum (Mo) Selenium (Se)
0.79–1.39 50–83
Sinkovič et al. (2021), Verma et al. (2020), Shreeja et al. (2021) and Podolska et al. (2021) Sinkovič et al. (2021) Sinkovič et al. (2021) Sinkovič et al. (2021), Podolska et al. (2021), Zhang and Xu (2017), Verma et al. (2020) and Thakur et al. (2021) Sinkovič et al. (2021), Podolska et al. (2021), Zhang and Xu (2017), Verma et al. (2020) and Thakur et al. (2021) Sinkovič et al. (2021) Sinkovič et al. (2021), Podolska et al. (2021), Zhang and Xu (2017), Verma et al. (2020) and Thakur et al. (2021) Sinkovič et al. (2021), Podolska et al. (2021), Zhang and Xu (2017), Verma et al. (2020) and Thakur et al. (2021) Sinkovič et al. (2021) Zhang and Xu (2017) and Verma et al. (2020)
consumption of buckwheat. It acts as a cofactor enzyme for many other nutrients increasing its bioavailability. Zhu (2016) explained that carotenoids present predominantly in the form of β-Carotene providing health benefits associated with carcinogenic diseases such as cardiovascular disease, macular degeneration, cardiovascular disease, UV induced skin damage, and cataracts. The other forms of carotenoids present in buckwheat are α-carotene, 9-cis-β-Carotene, 13-cis-β- Carotene, β-cryptoxanthin, and zeaxanthin. The dense source of B complex vitamin accumulated in highly in the regions of bran, followed by peripheral regions of endosperm and embryo (Semjon et al., 2021). Bastida et al. (2015) and Verma et al. (2020) demonstrated the forms of B complex vitamins namely Thiamine (vitamin B1) with 0.19–4.60 (mg/g), riboflavin (vitamin B2) with 0.55–1.4 (mg/g), niacin (vitamin B3) with 2.61–18.00 (mg/g), pantothenic acid (vitamin B5) with 11 μg/g, and pyridoxine (vitamin B6) with 1.5 μg/g. Daily dose of 6% pyridoxine shows effective action in the reduction of blood plasma homocysteine levels, folate which is considered as the most required
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Table 3.6 Vitamins present in buckwheat and its sprouts Buckwheat sprouts (mg/100 g)
Nutritional composition Vitamin E
Buckwheat seeds (mg/100 g) 1.3–16.4
α-Tocopherol
0.13
0.13–4.55
γ-Tocopherol
0.43
0.44–23.83
δ-Tocopherol
0.04
0.04 0.88
Vitamin C
7.10–13.05
13.75–21.39
β-Carotene
0.2–21
0.09–2.1
References Zhang and Xu (2017) and Verma et al. (2020) Sim et al. (2020) and Jeong et al. (2018) Sim et al. (2020) and Jeong et al. (2018) Sim et al. (2020) and Jeong et al. (2018) Verma et al. (2020), Bastida et al. (2015) and Sim et al. (2020) Verma et al. (2020), Sim et al. (2020) and Jeong et al. (2018)
nutrient for pregnant women because it helps in the generation of new cells which helps in healthy baby development (Nepali et al., 2019). The studies of Yiming et al. (2015) revealed that vitamin content increased about 3.5 times after germination period of 7 days. The amount of various vitamins present in different cultivars of edible buckwheat seeds and its sprouts discussed in the following Table 3.6.
3.3.2 Functional/Bioactive Composition of Buckwheat and Its Sprouts The world-wide dispersed buckwheat is cultivated and consumed in many regions due to the presence of bioactive compounds such as flavonoids, phenolic acids, phytosterols, fatty acids, amino acids and fagopyrins (Sytar et al., 2018). 3.3.2.1 Amino Acids The amino acid present in buckwheat is composed in well balanced form with high nutritional profile compared to other pseudo crops. Semjon (2021) discussed about the Osborne fractions associated with its amino acids. The four major Osborne fractions namely albumin, globulin, prolamin and glutelin. Albumin containing higher proportion of histidine, threonine, valine, phenylalanine, leucine, isoleucine, and lysine; globulin shows higher concentrations of methionine and lysine; prolamin encompassing with maximum percentage of histidine, threonine, valine, leucine, and isoleucine and finally glutelin was found to be rich in threonine, histidine, valine, leucine, and isoleucine.
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The most essential amino acids such as lysine, threonine, valine, arginine, and aspartic acid present in higher contents and lower concentrations of glutamine and proline was found in buckwheat rather than other cereals and wheat comparatively (Verma et al., 2020). Li (2019) explained that the first limiting amino acids such as arginine and lysine and second limiting amino acids include threonine and methionine. The low ratios of lysine/arginine and methionine/glycine have strong correlation in lowering the risk of cholesterol and hyperlipidemic effect. Nalinkumar and Singh (2020) stated that these first and second limiting amino acid also contribute for high digestibility with 80% digestibility coefficient. It results in increasing the biological value (BV) compared to true cereals such as wheat, rice, barley, and oats. The amino acids are accumulated in the outer shell of seed and micro elements such as Fe, Zn and K is rich in albumin and Ca, Mg, and Mn is greater in globulin and Na is found in prolamin and glutelin. Buckwheat can also be used as complementary food source to meet out the essential amino acids which lacks in cereals like lysine (Nepali et al., 2019). The list of essential amino acid along with its proportion is tabulated in the Table 3.7.
Table 3.7 Amino acid composition of buckwheat grains and its sprouts Nutritional composition Buckwheat Non-essential amino acids Alanine (%) 3.0–6.36 Aspartic acid (%) 5.2–11.80 Cystine (%) 0.15–0.18 Glutamic acid (%) 9.7–19.38 Glycine (%) 34.2–8.40 Proline (%) 2.6–7.93 Tyrosine (%) 1.5–3.03 Serine (%) 2.4–10.54 Essential amino acids Threonine (%) 1.9–4.04 Valine (%) 3.42–7.05 Isoleucine (%) 2.6–5.35 Leucine (%) 2.8–8.71 Methionine (%) 0.99–2.3 Phenylalanine (%) 2.0–6.08 Tryptophan (%) 1.5–2.14 Semi-essential amino acids Histidine (%) 1.4–5.39 Arginine (%) 5.4–13.18 Total amino acids 118.12 (mg/100 g DW)
Buckwheat sprouts 0.56–0.78 1.03–1.28 0.04–0.11 1.87–2.29 0.64–0.73 0.31–0.52 0.29–0.39 0.58–0.68 0.43–0.59 0.52–0.80 0.45–0.62 0.80–1.11 0.09–0.19 0.53–0.63
0.30–0.42 0.90–1.15
References Nepali et al. (2019), Deng et al. (2015), Zhang and Xu (2017) and Yiming et al. (2015)
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3.3.2.2 Phytosterols and Fatty Acids Buckwheat is a rich source of fatty acids such as palmitic, oleic, linoleic, arachidonic, stearic, behenic and lignoceric acid (Nalinkumar & Singh, 2020) (Table 3.8). Tömösközi and Langó (2017) expressed the major fatty acids present in buckwheat such as palmitic (C16:0) with 53.8%, oleic (C18:1) with 39.5% and linoleic (C18:2) acids with 27.9% contributing in total 95% of unsaturated fatty acids. In total, 40% is composed of polyunsaturated fatty acids (PUFA) which is considered as therapeutic component to decrease the risk of heart diseases. Li (2019) quantified the accumulation of fatty acid in embryo around 6.4% followed by hull around 0.4–0.9% and the least is found in endosperm around 0.2%. Semjon (2021) described the stability of fatty acids present in buckwheat. The composition of fatty acid remains constant and stable even after the storage period of 25 days with negligible change. Yiming et al. (2015) stated that contents of fatty acid such as palmitic acid, stearic acid, eicosenoic acid and arachidic acid have increased and linoleic acid contents have diminished. 3.3.2.3 Phenolic Compounds Phenolic compounds are phytochemicals composed of secondary metabolites, ubiquitous in plant materials. The structural configuration of polyphenol comprises of multiple hydroxyl groups (-OH) attached to one or more benzene rings within a molecule (Li, 2019). Polyphenolic compounds include flavonoids, anthocyanins, phenolic acids, anthraquinones, phenylpropanoid glycosides, and carotenoids. Polyphenols associated with various pharmacological properties such as antioxidant, antibacterial, antiadhesive and anti-inflammatory properties (Zhang et al., 2020) and provides physiological health benefits such as reducing the prevalence of chronic diseases like cardiovascular diseases, neurological disorder, and cancer (Zhu, 2016). Livadariu and Maximilian (2017) stated that the polyphenols act as a Table 3.8 Fatty acid composition of buckwheat grains Fatty acid composition Palmitic acid (C16:0) Stearic acid (C18:0) Oleic acid (C18:1) Linoleic acid (C18:2) Linolenic acid (C18:3) Arachidic acid (C20:0) Gadoleic acid (C20:1) Behenic acid (C22:0) Erucic acid (C22:1) Lignoceric acid (C24:0)
Quantity 12.4–18.6 1.2–2.3 35.9–39.5 34.4–39.7 1.4–2.6 0.9–1.5 1.9–3.0 1.4–1.5 0.2–0.6 0.6–0.9
References Verma et al. (2020) and Semjon (2021)
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plant protective agent against pathogens, ultraviolet radiation and microbes and the overall phenolic content was found to be increased after the germination period. 3.3.2.4 Flavonoids Flavonoids are the natural antioxidants, secondary metabolites, phytonutrients which have physiological health benefits such as reducing the risk of cardiovascular diseases, helps in reducing blood cholesterol level, blood pressure and microcirculation improver (Fu et al., 2020). Nam et al. (2018a, b) described the pharmacological properties such as anti-ageing, anti-tumour, and anti-inflammatory effects on human body. They are identified ubiquitously in plant species with working mechanism involved in plant growth and development, pigment generation, protective action against high light, ultraviolet radiation, and pathogens (Park et al., 2017). Recent findings exhibit that nearly 30 flavonoid compounds were found in the buckwheat. The basic chemical structure consists of 15 skeleton carbon compounds in the arrangement of (C6-C3-C6) (Jing et al., 2016). They belong to large group of benzo- γ-pyrone derivatives which are synthesized in the phenylpropanoid pathway with the production of various subclasses of flavonoids. The initiation of pathway occurs with the production of cinnamic acid with the aid of the enzymatic activity. The enzymes involved in the conversion of phenylalanine to secondary metabolites are phenylalanine ammonialyase (PAL), chalcone-synthase (CHS), chalcone isomerase (CHI), dihydroflavonol-reductase (DFR) and glycosyltransferases (GT) (Qin et al., 2017). The classification of flavonoids based on their molecular structure and their aromatic rings containing double bonds are responsible for their physical properties (Li, 2019). The main classification of flavonoids includes flavones, flavanols, isoflavones, flavonols, flavanones and flavanonols. According to glycosidic linkage pattern, they may either contain O-glycosides or C-glycosides. The bonding between sugar and hydroxyl group of flavonoid aglycone represents O-linkage and bond formation between anomeric carbon of sugar and carbon of flavonoid skeleton (C-C bond) represents C-linkage (Jang et al., 2021). Mansur et al. (2022) classified C-glycosyl flavones into four main components namely orientin (luteolin 8-C-glycoside), isoorientin (luteolin 6-C-glycoside), vitexin (apigenin 8-C-glycoside) and isovitexin (apigenin 6-C-glycoside). Rutin (quercetin 3-O-rhamnoside), quercetin, isoquercetin, and quercetin-3-O- robinobioside comes under the category of flavonol O-glycosides (Verma et al., 2020). Flavan-3-ol compound include catechin, epicatechin, epicatechin-3-O-p-hydroxybenzoate, epicatechin-3-O-(3,4-di-O-methyl)-gallate and catechin-7-O-glucoside. The composition of flavonoids, phenolic compounds, anthocyanins, and phenolic acids are mentioned in the following Table 3.9. The quantity and composition of phenolic compounds are influenced by several factors such as soil location, climatic changes, growth stages, seed composition, testa, size, and shape of the seed (Nalinkumar & Singh, 2020), geographical origin and growth phase of the plant,
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Table 3.9 Bioactive components present in buckwheat sprouts Bioactive components Total phenolic compounds (mg GAE/100 g F.W.) Total flavonoid compounds (mg/100 g QE F.W.) Flavone Orientin (mg/g DW)
Buckwheat Buckwheat grains sprouts References 298.96–365.67 66.4–490.07 Sturza et al. (2020), Beitane et al. (2018), Sim et al. (2020) and Wiczkowski et al. (2016) 47–275.58 72.71– Sim et al. (2020), Nam et al. 211.73 (2018a, b) and Sturza et al. (2020) 0.04–8.90
Isoorientin (mg/g DW)
0.17–16.67
Vitexin (mg/g DW)
0.18–9.84
Isovitexin (mg/g DW)
0.15–17.90
Flavonol Quercetin (mg/100 g DW)
5.00–9.71
Quercetin – 3-O robinobioside (mg/g DW)
0.02–4.56
Rutin (mg/g DW)
0.12–29.16
Isoquercetin (mg/g DW) Flavan-3-ol Catechin (mg/g DW)
0.04
Epicatechin (mg/100 g DW) Phenolic acids 4 hydroxybenzoic acid (μg/g DW) Chlorogenic acid (μg/g DW)
Caffeic acid (μg/g DW)
Nam et al. (2018a, b), Jang et al. (2019), Mansur et al. (2022) and Sim et al. (2020) Nam et al. (2018a, b), Jang et al. (2019), Mansur et al. (2022) and Sim et al. (2020) Nam et al. (2018a, b), Jang et al. (2019), Beitane et al. (2018), Mansur et al. (2022) and Sim et al. (2020) Nam et al. (2018a, b), Jang et al. (2019), Mansur et al. (2022) and Sim et al. (2020) Beitane et al. (2018) and Mansur et al. (2022) Nam et al. (2018a, b), Jang et al. (2019), Park et al. (2017) and Mansur et al. (2022) Nam et al. (2018a, b), Jang et al. (2019), Beitane et al. (2018), Park et al. (2017, 2019), Mansur et al. (2022), Sathasivam et al. (2021) and Sim et al. (2020) Mansur et al. (2022)
0.06–0.97
Beitane et al. (2018), Park et al. (2017, 2019) and Mansur et al. (2022) 15.50–20.64 Beitane et al. (2018) 1.79–84.85
Park et al. (2017, 2019) and Sathasivam et al. (2021) 54.13–163.9 Park et al. (2017, 2019), Mansur et al. (2022), Sathasivam et al. (2021) and Wiczkowski et al. (2016) 9.04–115.63 Park et al. (2017, 2019), Mansur et al. (2022) and Sathasivam et al. (2021) (continued)
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Table 3.9 (continued) Bioactive components Gallic acid (μg/g DW) Syringic acid (mg/g DW) p-Coumaric acid (mg/g DW) Ferulic acid (mg/g DW) Sinapic acid (mg/g DW) Ellagic acid (mg/g DW) Trans-cinnamic acid (mg/g DW) γ-Aminobutyric acid Anthocyanins Cyanidin-3-O-galactoside (mg/g DW) Cyanidin 3-O-galactopyranosyl- rhamnoside (mg/g DW) Cyanidin-3-O-glucoside (mg/g DW) Cyanidin-3-O-rutinoiside (mg/g DW) Proanthocyanidins (μg/g DW)
Buckwheat grains
Buckwheat sprouts 5.61–12.17
0.006–0.34 0.003 0.08 0.04–0.10
References Park et al. (2019) and Mansur et al. (2022) Mansur et al. (2022) Mansur et al. (2022), and Sathasivam et al. (2021) Mansur et al. (2022) Mansur et al. (2022) Mansur et al. (2022) Sathasivam et al. (2021)
3.65–6.03
Sim et al. (2020)
0.2–0.11
Mansur et al. (2022)
0.1–1.57
Mansur et al. (2022)
0.01–0.05
Mansur et al. (2022) and Seo et al. (2015) Mansur et al. (2022) and Seo et al. (2015) Wiczkowski et al. (2016)
0.007 0.006–1.2
0.02–0.11 15.7–20.7
plant tissue, maturation cycle, harvesting period and processing method (Vollmannova et al., 2021). Rutin (quercetin-3-rutinosid) is a representative and predominant flavonol glycoside present in buckwheat with significant functional properties (Tömösközi & Langó, 2017). It is composed of flavonol quercetin and disaccharide rutinose which has high antioxidant capacity due to the ability to generate reactive oxygen. Kumari and Chaudhary (2020) exhibits the pharmacological activities including antioxidant, anti-diabetic, anti-inflammatory, anti-neuroprotective, cytotoxic effects, cardiac hypertrophy inhibition, antihypertensive, antiplatelet aggregation, and anti-asthmatic effects. Quercetin-3-O-robinobioside have similar structure to rutin comprises of galactose-rhamnose attached to quercetin (Nam et al., 2018a, b). It possesses with functional properties like antioxidant, anti-inflammatory, anti-carcinogenic, and antihypertensive effects. Bastida et al. (2015) states that quercetin known as aglycone of rutin have immense antioxidant potential. This compound shows higher antioxidant activity than rutin and buckwheat seeds are the only pseudocrop variety to be contained with quercetin. Common buckwheat sprouts quantified for the presence of predominant flavonoids using quadrupole of-fight mass spectrometer (Q-TOF/MS). The results of
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Mansur et al. (2019) exhibit the components with respect to its peak in chromatogram. The components identified were luteolin-C-glucoside (orientin and isoorientin) at m/z 357.0614, vitexin at m/z 431.0983, isovitexin at m/z 431.0981, rutin at m/z 609.1464. 3.3.2.5 Phenolic Acids Phenolic acids determined in buckwheat seeds are chlorogenic acid, syringic acid, p-coumaric acid, ferulic acid, sinapic acid, caffeic acid, gallic acid, ellagic acid. The most predominant phenolic acid found in buckwheat is chlorogenic acid with protective action against degenerative disease, ageing, and blood pressure. Wiczkowski et al. (2016) describes the concentration of phenolic acids accumulated in different portions of buckwheat like cotyledons are enriched with cinnamic acid, o-coumaric (2-hydroxycinnamic) and synaptic (4-hydroxy-3,5-dimethoxycinnamic) acid; 3,4-dihydroxycinnamic acid is present in large quantities in hypocotyl region. Gan et al. (2017) describes that γ-Aminobutyric acid (GABA) is a non-protein amino acid synthesized from L-glutamic acid to GABA with the application of enzyme called glutamate decarboxylase (GAD), a pyridoxal 5′-phosphate dependent enzyme and the intermediate compound polyamine cane be converted into γ-aminobutyraldehyde via diamine oxidase (DAO) enzyme. This bioactive component is known for its depressive neurotransmitter activity, regulation of blood pressure and heart rate, increase insulin secretion from pancreas, helps to relieve pain and anxiety (Sim et al., 2020). The phenolic acid content increased up to 125.32% in buckwheat sprouts. The superior activity of phenylalanine ammonia-lyase (PAL) enzyme along with de novo biosynthesis of phytochemicals altogether enhances the phenolic acid content proportion (Thakur et al., 2021). The highest flavonoid content of 1090.25 mg/100 g DW and 221% of D chiro inositol was examined by Wang et al. (2013) while performing germination at 25 °C. Many studies reveals that at this temperature is considered as the most suitable temperature for germinating buckwheat sprouts. The phenolic compounds have positive correlation with antioxidant activity i.e. directly proportional to each other. The correlation of TPC explained by Wiczkowski et al. (2016) which is highest with Trolox equivalent antioxidant capacity (TCEA) followed by oxygen radical absorbance capacity (ORAC), Photoluminescence – antioxidant capacities of water soluble (PCL-ACW), DPPH assay. In buckwheat sprouts, maximum antioxidant activity was studied in TCEA and ORAC which is trifold times higher in cotyledons than hypocotyls followed by PCL-ACW which is double the time greater in cotyledon. 3.3.2.6 Anthocyanins Anthocyanins are the water-soluble pigments yet important bioactive component having highest antioxidant potential. They are categorized into subclasses of anthocyanins such as Cyanidin-3-O-galactoside, Cyanidin 3-O-galactopyranosyl-
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rhamnoside, Cyanidin-3-O-glucoside, Cyanidin-3-O-rutinoiside and proanthocyanidins (Mansur et al., 2022). Seo et al. (2015) demonstrated the biosynthesis pathway of anthocyanins where L-phenylalanine, primary precursor gets converted to glycosides of anthocyanidins by forming intermediate product such as dihydrokaempferol. 3.3.2.7 Mode of Action on Bioactive Compounds Accumulation There is a biochemical pathway leading to increasing the secondary metabolites such as phenols and flavonoids in germinated seeds. Sim et al. (2020) describes that during germination two major enzymes namely phenylalanine ammonia lyase (PAL) and tyrosine ammonia lyase (TAL) are important signal transducers taking part in shikimic acid-phenylpropanoid metabolism converting L-phenylalanine to trans-coumaric acid and L-tyrosine to p-cinnamic acid respectively. These metabolic products are responsible for the accumulation of higher number of bioactive compounds. Chen et al. (2022) explains that the products of p-cinnamic acid namely p-coumaroyl-CoA and malonyl-CoA leads to the production of 4,2′,4′,6′-tetrahydroxychalcone and 4,2′,4′-trihydroxychalcone. In the presence of enzymatic catalase such as chalcone synthase (CHS) and chalcone reductase (CHR) reacts on 4,2′,4′,6′-tetrahydroxychalcone to yield the secondary metabolites like flavone, flavonol, and flavanol. Flavanone is generated by the action of chalcone isomerase (CHI) on 4,2′,4′-trihydroxychalcone.
3.4 Antinutritional Properties Antinutritional compounds are present widely in outer layer (seed coat) of cereals, pulses, and legumes to prevent the food from external predators like insects, fly, birds, and animals. It has properties like protein agglutination, mineral binding capacity, complex formation with starch and in consumption causes digestion impartment in stomach and metabolic disrupt in small intestine (Deng et al., 2015). Bobkov (2016) stated that true digestibility of buckwheat is 80% which is lesser than cereals and the protein absorption is hindered by the presence of antinutritional compounds namely phytic acid, tannins, trypsin, or protease inhibitor and saponins.
3.4.1 Phytic Acid Phytic acid is a (myo-inositol 1,2,3,4,5,6-hexakisphosphate, InsP6) the main reservoir of phosphate, present in the form of phytate salts, an antinutrient compound found in the outer layers of seed ranging from 1% to 3% dry matter. It chelates with bivalent minerals like calcium, copper, iron, and zinc, energy providing molecules
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such as starch, protein and enzyme result in complex molecule formation making it unavailable for human absorption and assimilation (Mattila et al., 2018a, b). Bobkov (2016) described that evenly distributed phytic acid is present in vacuolar derived protein bodies, accumulated highest in embryo and aleurone layers ranging from 0.21% to 0.72% and maximum found in higher concentrations in buckwheat compared to legumes and cereals. Dulinski et al. (2020) it chelates with Ca2+ ions and cofactors like Mg2+ and Zn2+ resulting in inhibition of α-amylase and protease activity. Cankurtaran and Bilgiçli (2021) determined the phytic acid of 633.80 mg/100 g in raw buckwheat flour. During germination, hydrolysis of phytic acid occurred due to the higher rate activity of phytase. The hydrolysis results in the depletion of the phytic acid content by 17.42% through the conversion of phytic acid into myoinositol and phosphoric acid (Thakur et al., 2021).
3.4.2 Condensed Tannins Condensed tannins (CT) composed of complex molecules containing group of both beneficial and non-beneficial phenolic compounds. They are capable of binding predominantly with protein and starch result in the reduction of nutritional value of food (Mattila et al., 2018a, b). Tannin is concentrated in bran layer than in endosperm region. Bobkov (2016) mentioned that higher concentration of 6% present in bran and embryo and lowest concentration of 0.3% and 0.1–0.3% found in hull and flour proportions. The most significant method to reduce the tannin content is germination which depleted 59.91% under 72 h treatment. The repulsive interaction caused by hydrophobic bonding between polyphenolic compounds and organic compounds (proteins, carbohydrates) leads to the reduction of tannin content (Thakur et al., 2021).
3.4.3 Trypsin Inhibitor Trypsin inhibitor is the antinutrient present widely in cereals, legumes, and pseudo cereal like buckwheat. Their working mechanism is to inhibit the activity of trypsin resulting in poor digestion and metabolism of protein molecules (Zhang et al., 2015). Ungerminated seed was found to have 51.34 U/g but after the first day of germination, Wang et al. (2013) examined 61% reduction containing 19–22 U/g of trypsin inhibitor.
3.4.4 Methods to Remove Anti-nutritional Compounds The removal of antinutritional compound is an important criterion to improve the better utilization of crop cereals by deactivating the antinutrient compounds, increasing bioavailability, improve nutritional value and digestibility. The most
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common method used is boiling, soaking and germination and in recent days, microwave heating and high hydrostatic pressure techniques are widely used (Deng et al., 2015). Dulinski et al. (2020) stated the methods such as thermal hydrolysis and enzymatic activity for phytate dephosphorylation. Three major applicable enzymes are 3-phytase A (E.C.3.1.3.8), 6-phytase A (E.C.3.1.3.26) and phytase B (E.C.3.1.3.2)
3.5 Effect of Different Treatments on Growth and Chemical Composition of Buckwheat Sprouts 3.5.1 Effect of Laser Light Treatment The laser light of He-Ne source illuminated on soaked seeds at 632 nm which had a positive impact on the germination efficiency. Almuhayawi et al. (2021) findings show that higher biomass accumulation, fresh and dry weight, higher seed size was observed in common buckwheat sprouts exposed to laser beam. The light absorbed by phytochromes increases the chemical energy required for the cell lysis, and activation of enzymes like protease ad amylase to yield both quantitative and qualitative sprouts. Indirect means of improving germination is photosynthetic pathway, chlorophyll and carotenoid components aids in this process. In his study, significant amount of carotenoids in specific lutein followed by α-carotene played a predominant role in the growth of sprouts. Elevation in macro minerals such as K, P and Na was found along with higher phenolic and flavonoid content. The greater the phenolic content, higher the antioxidant activity. Phenolic compounds such as rutin, predominant flavonoid which has higher antioxidant potential followed by catechin and p-coumaric acid were estimated in three-fold number in laser treated sprouts. The presence of sinapic acid and isovertexin; velutin was found in tartary and common buckwheat sprouts respectively. The precursor of the anthocyanin, L-Phe used in treating the seeds for germination which have shown 30% increase in predominant anthocyanin named rutin. Seo et al. (2015) declared that optimum concentration of 5 mM shows the maximum anthocyanin accumulation in sprouts, higher concentration deteriorates the biosynthesis of rutin. Higher content of phenolic acid, flavonoid content, and vitexin was accumulated under red blue light and maximum accumulation of chlorogenic acid and rutin found in red, blue, and white light. Comparing the growth conditions under darkness, double the amount of anthocyanin increased under white light. Comparative analysis of buckwheat with and without testa performed under 3 different lights namely white, red, and blue by Livadariu and Maximilian (2017). The germinated seeds with testa and without testa have shown better growth characteristics such as sprout rate, fresh weight of sprouts, cotyledons, root, hypocotyls, and its length under red light in exception fresh weight of cotyledons (seeds without testa) had better value under blue light. The bioactive components such as
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polyphenols, flavonoids and its associated antioxidant activity was found higher quantity in seeds with and without testa under red and blue light. The highest shoot and root length was examined under red and fluorescent light respectively as per the observations of Nam et al. (2018a, b). The effect of blue light abundantly increases the polyphenols double the quantity, flavonoids by triple the amount and the accumulation of its subclasses such as rutin ranks first followed by C-glycosylflavones (isoorentin and isovitexin), vitexin, quercetin-3-O- robinobioside, and orientin. Shorter wavelength (460 nm) of blue light with cryptochromes responsible for the gene expression of flavonoid synthesis (chalcone synthesis and glycosylation of aglycones) and phototropins together contributes for the higher accumulation of flavonoid and phenolic components. Growth characteristics such as shoot and root length, fresh and dry weight of sprouts examined by Nam et al. (2018a, b) and he states that better morphological features observed under red light. Fluorescent and blue light improves the accumulation of flavonoid such as vitexin, isovitexin, orientin, isoorientin and rutin respectively.
3.5.2 Effect of Ultrasound Assisted Solvent Extraction The implementation of green technique is an emerging technique in that case ultrasound assisted DES (deep eutectic solvent) extraction method was equipped by Mansur et al. (2019). The DES solvent sprout extracts in specific 80% CCTG-DES (Choline Chloride Triethylene Glycol) have reported for the highest flavonoid content with imbibition of 20% water (w/v). The addition of water decreases the viscosity, increases the polarity which enhances the elution of components. Either inclusion of higher water amount or exclusion of water decreases the elution efficiency. Usage of 80% CCTG-DES implies the highest storage stability of 80% of the eluted components such as orientin, isoorientin, vitexin and isovitexin.
3.5.3 Effect of Sodium Bicarbonate Solution Salinity stress is one of the abiotic stresses implied to improve the function metabolites during germination. Sodium bicarbonate (NaHCO3) was applied by Qin et al. (2017) in his experimentation and studied the accumulation of bioactive compounds. There is a correlation between accumulation of phenolic compounds (29.31 ± 0.45 mg/g), flavonoidssuch as rutin, quercetin, isoquercetin, and kaempferol (26.69 ± 0.40 mg/g), free D-chiro-inositol (DCI) (766.21%) and α-glucosidase inhibitory activity (180.09%) was examined highest at the lower NaHCO3 concentration of 0.05% after 96 h of germination.
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3.5.4 Effect of Metallic Additives The metal additive solutions of Al3+, Cu2+, and Zn2+ have increased the flavonoid content by 1315.52 mg/100 g DW at 1000 mg/L, 30% higher than control at 0–60 mg/L, and 1189.42 mg/100 g DW at 500 mg respectively. The findings of Wang et al. (2013) declared that all these metal additives have higher efficiency at lower concentrations. When the additives concentration elevates, it depreciates the enzymatic activities of phenylalanine ammonia lyase (PAL) and chalcone isomerase (CHI) involved in phenylpropanoid pathway resulting in lesser accumulation of bioactive compounds. As trypsin inhibitor is thermostable, there is 25% trypsin inhibitor (TI) retention in processed flour. These metal additives reduce TI concentration exponentially, comparatively Al3+ have highest reduction capacity at 1000 mg/L from 51.34 to 4.90 U/g of TI.
3.5.5 Effect of Plant Growth Hormones The treatment with four different growth regulators such as 6-benzyl amino purine (BAP), kinetin, zeatin, and thidiazuron (TDZ) were applied to seeds by Sathasivam et al. (2021). Among those lower concentrations of 0.1 mg/L of BAP reported for the highest fresh weight (g), root and shoot length (cm) of sprouts and the lowest observations found in TDZ. BAP is considered as one of the best growth regulators and as significant cytokinin responsible for the morphogenic aptitude in plant growth. 0.1 mg/L of TDZ was reported for the highest accumulation of total phenolic contents and 3 important flavonoids namely rutin, caffeic acid, 4-hydroxybenzoic acid followed by 1 mg/L of kinetin, BAP and zeatin. Conclusions of Park et al. (2017) reveals that lowest concentration of 0.5 mg/L of gibberellic acid (GA) followed by 3-indoleic acid (IAA) reported for the highest fresh weight, root and shoot length. Negative impact was observed in the usage of higher dose. Highest total phenolic content of 1580.49 ± 11.19 μg/g DW was observed at 0.1 mg/L of IAA, highest peak value of catechin and caffeic acid at 0.5 mg/L of IAA and rutin at 1.0 mg/L of IAA followed by 0.1 mg/L of GA. Auxins such as IAA and GA are important phytohormones responsible for cell division, elongation, differentiation and plant growth and development respectively. The aforementioned statement proves the results of Park et al. (2017) that GA and IAA have shown highest growth characteristics and phenylpropanoid accumulation respectively.
3.5.6 Effect of Elicitors Highest phenolic content and 6 phenolic compounds such as caffeic acid, chlorogenic acid, gallic acid, rutin, catechine, (−) epicatechine were observed by Park et al. (2019) in the germinated seeds treated with 0.1% chitosan and 150 μM
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jasmonic acid at 72 h of germination. These elicitors create a stress which helps in the plant growth and development. Chitosan and jasmonic acid elicitation involves in the elevation of production of phenylpropanoids and increase the total phenolic content respectively.
3.5.7 Effect of Sucrose and Calcium Chloride The studies on sucrose treated germinated seeds was carried out by Jeong et al. (2018) exhibits that height and weight of sprouts was affected but in contrast phenolic acids and flavonoids namely orientin, isoorientin, vitexin, isovitexin, and rutin have increased in the range of 52–68% at 3% sucrose concentration. The observations of α-tocopherol, γ-tocopherol and vitamin C have found highest 4.55 mg/100 g, 23.83 mg/100 g, and 29.39 mg/100 g respectively at ideal 3% sucrose concentration and elevation of 85.7% of β-carotene at 5% sucrose level which have positive correlation with antioxidant activity. Secondary metabolites such as phenolic acid and flavonoids (orientin, isoorientin, vitexin, isovitexin and rutin) was found its maximum of 490.07 ± 6.92 GAE mg/100 g FW and 182.88 ± 3.08 CE mg/100 g FW at 3% sucrose and 7.5 mM CaCl2. Maximum value of γ-aminobutyric acid was noticed at 3% sucrose solution. Sim et al. (2020) explained the synergistic effect between the combination of sucrose and calcium chloride. As sucrose depletes the plant growth by causing water deficiency, osmotic pressure and oxidative stress, calcium takes part in signal transduction combining both calmodulin and protein kinase results in regulating the plant growth. 3% sucrose elicitation showed highest free radical scavenging and lipid peroxidation inhibitory activity. In contrast DPPH, ABTS scavenging activities and reducing power reported its peak activity at 3% sucrose and 7.5 mM CaCl2. Both the treatments contribute the highest antioxidant activity.
3.5.8 Effect of Slightly Acidic Electrolysed Water (SAEW) Stress condition is provided to grains by treating with slightly acidic electrolysed water which improves γ-Aminobutyric acid (GABA). The significant increase in GABA was observed by Hao et al. (2016) which was 143.20 ± 5.91 mg/100 g much higher than control on 6th day of germination. After 6th day, insignificant decrease was observed. There is a relationship between activity of glutamic acid decarboxylase (GAD) and GABA. SAEW increases the GAD activity which directly elevates the GABA levels. Rutin, predominant flavonoid examined highest value of 739.9 ± 24.7 mg/100 g on 8th day of germination. SAEW increases the rutin content by preventing the conversion of rutin to quercetin which is catalysed by rutin degrading enzyme (RDE) and increases the PAL activity. In addition, it controls the bacterial contamination significantly by immersing the seeds in SAEW solution
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during germination of grains. Liang et al. (2019) studied the bacterial contamination on SAEW treatment on buckwheat grains. Microflora contamination is one of the major issues during germination because water plays a significant role. SAEW have shown positive and better results compared to tap water. The results of applying SAEW as disinfectant in germination have shown 0.60–1.30 log10 CFU/g incase of fungi namely yeast and mold, and 1.98–3.06 log10 CFU/g incase of total aerobic bacteria.
3.6 Agricultural Utilization The underutilized buckwheat is an ecological adaptable pseudo crop and, it withstands and grow at extreme environmental conditions. They produce high nutritive quality crop by utilizing lesser amount of fertilizers and nutrients with high efficiency and shows good weed suppression activity (Domingos & Bilsborrow, 2021). Buckwheat has diversified function in agriculture, it can perform as cover crop, break crop, smother crop, nutrient conserving crop, gourd crop, and green manure crop as shown in (Maurya et al., 2021).
3.6.1 Cover Crop Cover crop is considered as an important and efficient agricultural strategy which helps to withhold nutrient retention in soil, maintains the soil in fertile nature, reduce water and soil erosion, enhance soil health and its structure. Apart from providing these beneficiaries, it also used as weed suppresser in field crop and prevent the use of chemical fertilizer (Smith et al., 2020). Masilionyte et al. (2017) experimented using cover crop as buckwheat in crop rotation technique to have positive impact on weed suppression. The infusion of cover crop in crop rotation aids in reducing the yield loss of cash crop and increase the competency for light, nutrients, and space of weed, providing unfavorable growth conditions for weed. The mixed cropping of buckwheat along with white mustard have shown positive results in weed suppression. Lopes et al. (2021) studies reveal that intercropping improves the soil health and use of resource efficiency is on higher score. The combination of legume-cereal cropping pattern considered as the best method compared to others hence buckwheat and maize is used in the intercropping to enrich the nutrient content of soil. Babu et al. (2018) described that intercropping and crop diversification like maize cropping in uplands with buckwheat result in enhancing land profitability, make crop adaptable to climatic change and increase the farmer income. The weed suppression activity is achieved by the presence of 3 natural phytotoxins present in stems of buckwheat namely fagomine, 4-piperidone, and 2-piperidine. Tursun et al. (2018) tested the growth of buckwheat with apricot orchard. The apricot orchard
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yield was decreased by growth of weeds such as Euclidium syriacum (L.) R. Br., Veronica polita Fr., and Polygonum aviculare L causing plum pox virus. Compared to using weedicide or other chemicals, buckwheat has shown 73% contribution in weed suppression maintaining the fertility of soil.
3.6.2 Insect Pollinator Crop The crop pollination can be enhanced by cropping pollinators with wild flowering species. The findings of Nagano et al. (2021) revealed that buckwheat cultivation along the field margins of wild flowering species increased the crop pollination in exponential manner. Buckwheat is a self-incompatible species, distylous flower, acts an insect attractant which result in improving the pollination of wild flower. The insects such as bees, butterflies, hoverflies, and beetles get attracted abundantly to the flowers of buckwheat and this interaction results in pollinator sharing between pollinator and wildflower. The higher efficiency of pollination is seen in similar morphological structure between crop and pollinator. Fagopyrum esculentum sp., is a short duration and most importantly cultivated crop in mountain regions. It is a moisture loving annual crop grown preferably in cold climatic conditions especially in hilly regions above 1600 m high. Manhare and Painkra (2018) studied on the entamophilic honeybees as pollinators to observe the production yield of buckwheat and honey. The extended flowering period of buckwheat for 30 days results in the production of 50–100 kg of honey per hectare. The usage of bee attractants such as honey solution (10%) and jaggery solution (15%) have reflected the increased production of buckwheat.
3.6.3 Biological Fertilizer The left-over waste material such as buckwheat husk obtained after processing of buckwheat groat into flour and various products is rich in organic minerals. Pociene and Šlinkšienė (2022) studied on the formulation of granulated organic fertilizer from buckwheat husk. The buckwheat husk ash is composed of primary and secondary soil enriching nutrients such as P2O5, K2O, CaO, and MgO, and micronutrients such as Zn, Mn, Cu, Fe, Co, and Mo. The higher concentrations of major and minor nutrients are determined in fertilizer ranging from 35.92% water soluble potassium to 38.63% hydrochloric acid soluble potassium; CaO, and MgO was observed in the levels of 12.18% and 3.56% respectively. The bulk density determines the strength of fertilizer ranging from 6.18 to 7.39 N/granules with spherical shape. The granular spherical shape should be maintained by storing at 21–23 °C and 70–75% humidity. Ozyazici and Turan (2021) optimized and determined the nutrient composition of buckwheat fertilizer obtained by vermicomposting process. Vermicomposting involves modifying organic material into humus like substance by earthworms with
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additional help of microbes. The major soil nutrients such as N, P, and K ranging from 1.65–1.92%, 0.23–0.27%, to 0.75–0.84% have positive impact on physical, chemical, and biological attributes of soil. The secondary major nutrients such as Ca and Mg present in stable form varies from 0.03–0.06% to 0.20% respectively. The micronutrients containing 40 ppm of Fe, 33.7 ppm of Mn, 9.5 ppm of Cu, and 8.7 ppm of Zn.
3.6.4 Trap Crop Trap cropping is the technique to divert, attract or intercept the selected pest towards non-crop habitat to protect and prevent damage caused by insect to economically important cash crop (Shrestha et al., 2019). This tactic is an attractive and promising method to entrap and divert damage causing insects towards trap crop, insects consider trap crop as host which is cultivated in the same surface region closer to main cash crop. Sarkar et al. (2018) demonstrated that Thrips tabaci Lindeman (Thysanoptera: Thripidae) nutrient absorbing weed of onion (Allium cepa L. (Amaryllidaceae)) is controlled by cultivation of buckwheat along with the onion. For the effective trap cropping implementation, the crop should be attractive to the targeted insect and this green technique acts as a defendable and foolproof mechanism to insects. Buckland et al. (2017) worked to solve the complication of reduction in the yield of 60% onion caused by Iris yellow spot virus, a thrips vectored Tospovirus. The research findings on application of buckwheat as trap crop sorted out this worldwide complication by attracting and depletion of onion thrips adult.
3.7 Application/Utilization of Buckwheat The highly nutritious buckwheat is consumed as main course of meal and considered as a functional food source. Depending upon the habitat, culture, lifestyle and food pattern, the processing, cooking method and consumption of buckwheat varies in each and every region throughout the world. Patil and Jena (2020) described the global consumption of buckwheat utilized in different formulations, (Fig. 3.2) the raw buckwheat flour (kanchopitho) is incorporated in chapathi, noodles, and pancake. In Nepal, it is consumed in the form of tea, vinegar, jam, momo, sweet, dorpa dal, chhyang or jaand (beer), roasted grain, macaroni, porridge, sattu and pakauda. In Japan, the most consumable form of buckwheat is noodles followed by beer, locally prepared beverage called Chang. Zhou et al. (2015) buckwheat contains similar starch properties of corn, and it has an amylose content of 21–26% in overall 70% of starch content. The presence of starch appreciates the production of products such as cookies, biscuits, noodles, and pasta and they have high biological value due to the polymeric substances such as protein, starch, flavonoids, and other bioactive components.
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Bread
Bun
Sour dough
Biscuit
Cookie
Pancake
Cake
Bakery products Tea
Noodles Commercial products
Extruded products Pasta
Honey
Traditional products
Macaroni
Beer
Chappathi
Porridge
Sattu
Pakauda
Couscos Makgeolli
Dumpling
Fig. 3.2 Different products utilized from buckwheat
3.7.1 Bakery Products Farzana et al. (2021) studied on the incorporation of buckwheat flour in the cake formulated with wheat flour. The cake was prepared using wheat, 5 different concentrations of buckwheat from 0% to 40%, sugar, salt, yeast, and egg made into cake batter, baked at 170–180 °C for 20–30 min. The incorporation of buckwheat flour at 30% concentration was highly acceptable and it showed elevation in the nutritional composition of protein, fat, fiber, sodium, potassium, calcium, and iron from 10–14 g, 34–39 g, 0.27–0.35 g, 120–124 mg, 200–271 mg, 25–28 mg, to 3.6–5.09 mg respectively. Hussain et al. (2017) worked on designing gluten free food for celiac disease patients considering buckwheat for its high crude protein, high amino acid, high lysine content and gluten free content. He prepared most market demandable cookies using ingredients such as wheat flour, buckwheat flour, sugar, fat, salt, egg, and baking powder in desirable proportions. The prepared cookie with the incorporation of buckwheat flour to wheat flour was observed for its macro and micronutrients. The comparative analysis between wheat and buckwheat flour relies on the gradient of protein, fat and fiber increased from 12.53–15.79%, 1.42–1.81% to 0.70–1.93%.
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Bastida et al. (2015) reviewed on the incorporation of buckwheat flour into wheat bread. Numerous research findings reveals that the antioxidant capacity of bread incorporated with buckwheat flour was on higher range and was stable even after the storage period of 4 weeks. Buckwheat contains volatile aromatic compounds such as 2,5-dimethyl-4-hydroxy-3(2H)-furanone, (E,E)-2,4-decadienal, phenylacetaldehyde, 2-methoxy-4-vinylphenol, E-2-nonenal, decanal, hexanal and salicylaldehyde (Jing et al., 2016). These aromatic compounds and reducing sugars such as glucose, fructose, and maltose are responsible for attaining the highly acceptable organoleptic bread product.
3.7.2 Extruded Products Starowicz et al. (2018) analyzed the sensory evaluation of various extruded products such as spaghetti, pasta and noodles prepared with buckwheat flour. The prepared extruded products have better bulking properties, firm and dense structure, increased adhesiveness, high pasting property, highly acceptable organoleptic characteristics like color, texture, taste, flavour and aroma. The bioactive component, rutin was found significantly higher in the products prepared with buckwheat flour. Man et al. (2016) optimized the pasta preparation with infusion of buckwheat flour at different concentrations from 0% to 25%. The approach of buckwheat flour into pasta was considered due to its high nutritional and biological value, high amino acid profile, well balanced major and minor nutrients, and high antioxidant potential. The highly consumable pasta was prepared with wheat flour, buckwheat flour and egg. The product formulated with 25% buckwheat flour was found to be highly acceptable in terms of sensory attribute, nutritional profile, and physico- chemical property of pasta. Defries et al. (2018) conducted the comparative analysis to determine the satiety levels of snack product such as buckwheat groats and buckwheat pita against reference snack sample such as corn groat and rice bread respectively. These buckwheat snack products have beneficial satiety compounds which release gastrointestinal hormones and have higher crude protein and fiber content increase the satiety and helps in weight loss reduction. The organoleptic characteristics were found to be similar and the energy value of buckwheat snack product is moderately higher than reference sample.
3.7.3 Fermented Products Khan et al. (2017) formulated the thymorutin green tea with the combination of buckwheat (Fagopyrum esculentum) and wild thyme (Thymus serpyllum) leaves and flowers. The green tea is considered as a major source of antioxidant in recent days and consumed in daily diet. The tea is prepared by a sequential process of
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cleaning, washing, steaming, dehydration, chopping, addition of additives and packaging. The major antioxidant compound present in thymorutin green tea is rutin, which contributes around 1.99%, and additional nutrients such as caffeine with 0.35%, ethanol with 18.93%, thymol with 0.21%. Rutin increases strength of capillary walls, improve the dissolubility of more oxygen, increases elasticity of veins and thymol blood vessels. Suzuku et al. (2020) explained the production of vinegar with the buckwheat as raw ingredient. The process gets initiated with boiling of buckwheat with koji resulting in alcoholic fermentation followed by acetic fermentation. The resultant product of buckwheat vinegar stands out from other vinegar both nutritionally and organoleptically. It develops unique flavor and aroma due to the formation of aldehyde and ketone groups and contains abundant antioxidant and flavonoid compounds. Brasil et al. (2020) have attempted the replacement of barley malt with buckwheat malt in beer production. The malt is usually used as adjunct, and it determines the beer quality. The buckwheat is used as it has higher nutritional value and similar structural and chemical characteristics to cereals such as wheat and barley and cheap source of crop. The beer is produced using buckwheat adjunct in a serious of procedure and the properties of buckwheat mash reflected same as barley mash. The buckwheat malt as adjunct satisfied the quality attributes of beer and have improved antioxidant, polyphenolic content. 5-methylfurfural produced a spicy and sweet coconut like aroma which is developed during beer manufacturing.
3.7.4 Traditional Products Semjon et al. (2021) discussed the traditional foods prepared and consumed in different regions of the world, Tatarcanepirohy, commonly called as Tartar dumpling in Eastern part of Slovakia, Soba noodles (buckwheat noodles) in Japan, buckwheat pasta, cookies, and sourdough. Demir and Demir (2016), Couscous is a Turkish based traditional food prepared generally with wheat granules, durum semolina, wheat flour, milk. It is popular dish in Spain, Italy, Greece, and Portugal and industrially known as pasta. Buckwheat and legume flour was infused into the traditional Turkish food to reduce malnutrition in population. The resultant product has comparatively better cooking, physical and chemical properties than traditionally prepared wheat-based couscous. He suggested the preparation of flour-based products like couscous, noodles, pasta, and pancakes can be prepared with buckwheat flour or with combinations of legume flour rather than the use of wheat flour to enhance the nutritional quality of the product. Park et al. (2018), Makgeolli is a traditional alcoholic beverage of Korea. A traditional fermentation starter Nuruk composed of fungi, yeast and lactic acid bacteria is used in brewing along with starch and sugar. Buckwheat is used as starch source
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and fermented by R.oligosporus resulting in the formation of L-carnitine. L-carnitine has a therapeutic property to reduce the risk of heart disease.
3.7.5 Commercial Products Lodhi and Vadnere (2019) discussed the worldwide consumed energy drink and buckwheat infusion under the category of caffeinated beverages. The common ingredients involved in energy drink are taurine, pyridoxine, niacin, niacin, cyanocobalmin, riboflavin, glucuronolactone, inositol, caffeine, theobromine and theophylline, ginkgo biloba extract, yohimbine, ginseng extract, and sugars. The plant derived beverage have multitude nutritional and therapeutic value due to the presence of larger amount of rutin from 2% to 10%, predominant buckwheat flavonoid compound. Hussain (2018) formulated the instant porridge with underutilized crop as buckwheat. The buckwheat is brought into formulation considering its nutritional and functional properties namely high protein ratio, bioactive components, vitamins, and minerals, reduces and regulates blood pressure, maintains blood sugar level, prevent fat deposition, and lowers blood cholesterol respectively. The grits are processed in a sequential manner starting from preliminary process like cleaning and washing followed by roasting at 120 °C for 10 min, cooling, milling, and dehulling. The porridge can be prepared by reconstituting grits with milk and sugar is added as sweeteners. The study reveals that porridge was found with higher functional, physicochemical, and nutritional properties and observed that roasting in a processing step improves the nutritional value.
3.8 Conclusion Buckwheat is ubiquitous crop, belonging to the Polygonaceae family and Fagopyrum genus. They comprise of starchy endosperm and non-starchy fiber rich aleurone layer similar to cereals such as rice and wheat. Buckwheat sprout is composed of higher concentration of protein with improved protein digestibility, total and reducing sugars, micro and macro elements, vitamins like vitamin B1, B2, B6, E, and C, well balanced amino acid profile comparatively higher than buckwheat grains and stands out from other pseudocrops and cereals, due to the presence of health benefits providing compound such polyphenol, flavonoid such as orientin, quercetin, vitexin, isovitexin, and isoorientin and anthocyanidins. Sprouting is performed as a sequential operation of washing, soaking, and germinating under controlled growth chamber by maintaining ambient conditions and stress conditions provided by some elicitors, and abiotic compounds. The mode of action, biochemical pathways and enzymatic actions involved in rising the chemical composition of protein, amino acid, total sugars, phenolic acids,
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flavonoids in particularly rutin, and anthocyanins was discussed in detail. The effect of various treatments and stress factors on germinating seeds and the modifications observed in growth characteristics and functional composition, prevention from bacterial contamination was explained. Antinutritional compounds such as phytic acid, trypsin inhibitor, tannin, and saponin present in buckwheat which causes harmful effect and disturbs the physiological metabolism of human body and act as a barrier for the proper utilization and consumption by human population. There are several methods to deduce the concentration of antinutrients such as smoking, boiling, roasting, and germination. Among these methods, germination is considered as the ideal method as it decreases the antinutrient content and proliferates the proportion of nutritional and functional compounds. The energy value, protein, fat, and fiber are found to in dense quantities in buckwheat sprouts. The major flavonoid component present in buckwheat is rutin and quercetin since it has an abundant antioxidant potential and due to the presence of these bioactive compounds, buckwheat is promoted as a functional food source. Considering this functional and nutraceutical properties this crop attains recognition, consumed by various countries, and industrially commercialized as tea, beer, honey, instant porridge mix, fermented beverages, and energy drink. The crop is considered as farmer’s friendly crop since it grows in extreme climatic conditions, grows, and utilizes using lesser amount of fertilizer with absorption of higher efficiency of nutrients, the waste obtained after processing can be converted into organic fertilizer, forage crop, trap crop, insect pollinator crop and so on. The multipurpose crop with multitude properties considered as functional food crop and agronomically important crop. It should be consumed and utilized in wide range to prevent the exploitation and to enhance the human health.
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Sarkar, S. C., Wang, E., Wu, S., & Lei, Z. (2018). Application of trap cropping as companion plants for the management of agricultural pests: A review. Insects, 9(4), 128. https://doi.org/10.3390/ insects9040128 Sathasivam, R., Kim, M. C., Chung, Y. S., & Park, S. U. (2021). Effect of cytokinins on growth and phenylpropanoid accumulation in Tartary buckwheat sprouts (Fagopyrum esculentum Moench). Journal of Aridland Agriculture, 7, 89–94. https://doi.org/10.25081/jaa.2021.v7.7198 Semjon, B. (2021). Buckwheat as vanished plant ingredient in traditional meals in the eastern part of Slovak Republic. In Indicators of change in cultural heritage. Wydawnictwo Uniwersytetu Rolniczego im. Hugona Kołłątaja. Seo, J. M., Arasu, M. V., Kim, Y. B., Park, S. U., & Kim, S. J. (2015). Phenylalanine and LED lights enhance phenolic compound production in Tartary buckwheat sprouts. Food Chemistry, 177, 204–213. https://doi.org/10.1016/j.foodchem.2014.12.094 Shreeja, K., Devi, S. S., Suneetha, W. J., & Prabhakar, B. N. (2021). Effect of germination on nutritional composition of common buckwheat (Fagopyrum esculentum Moench). International Research Journal of Pure and Applied Chemistry, 1–7. https://doi.org/10.9734/IRJPAC/2021/ v22i130350 Shrestha, B., Finke, D. L., & Piñero, J. C. (2019). The ‘botanical triad’: The presence of insectary plants enhances natural enemy abundance on trap crop plants in an organic cabbage agro- ecosystem. Insects, 10(6), 181. Sim, U., Sung, J., Lee, H., Heo, H., Jeong, H. S., & Lee, J. (2020). Effect of calcium chloride and sucrose on the composition of bioactive compounds and antioxidant activities in buckwheat sprouts. Food Chemistry, 312, 126075. https://doi.org/10.1016/j.foodchem.2019.126075 Singh, M., Malhotra, N., & Sharma, K. (2020). Buckwheat (Fagopyrum sp.) genetic resources: What can they contribute towards nutritional security of changing world? Genetic Resources and Crop Evolution, 67(7), 1639–1658. https://doi.org/10.1007/s10722-020-00961-0 Sinkovič, L., Sinkovič, D. K., & Meglič, V. (2021). Milling fractions composition of common (Fagopyrum esculentum Moench) and Tartary (Fagopyrum tataricum (L.) Gaertn.) buckwheat. Food Chemistry, 365, 130459. https://doi.org/10.1016/j.foodchem.2021.130459 Smith, R. G., Warren, N. D., & Cordeau, S. (2020). Are cover crop mixtures better at suppressing weeds than cover crop monocultures? Weed Science, 68(2), 186–194. https://doi.org/10.1017/ wsc.2020.12 Starowicz, M., Koutsidis, G., & Zieliński, H. (2018). Sensory analysis and aroma compounds of buckwheat containing products – A review. Critical Reviews in Food Science and Nutrition, 58(11), 1767–1779. https://doi.org/10.1080/10408398.2017.1284742 Sturza, A., Păucean, A., Chiș, M. S., Mureșan, V., Vodnar, D. C., Man, S. M., et al. (2020). Influence of buckwheat and buckwheat sprouts flours on the nutritional and textural parameters of wheat buns. Applied Sciences, 10(22), 7969. https://doi.org/10.3390/app10227969 Suvorova, G., & Zhou, M. (2018). Distribution of cultivated buckwheat resources in the world. In Buckwheat germplasm in the world (pp. 21–35). Academic. https://doi.org/10.1016/B978- 0-12-811006-5.00003-3 Suzuki, T., Noda, T., Morishita, T., Ishiguro, K., Otsuka, S., & Brunori, A. (2020). Present status and future perspectives of breeding for buckwheat quality. Breeding Science, 19018. https:// doi.org/10.1270/jsbbs.19018 Sytar, O., Biel, W., Smetanska, I., & Brestic, M. (2018). Bioactive compounds and their biofunctional properties of different buckwheat germplasms for food processing. In Buckwheat germplasm in the world (pp. 191–204). Academic Press. Takeshima, R., Nishio, T., Komatsu, S., Kurauchi, N., & Matsui, K. (2019). Identification of a gene encoding polygalacturonase expressed specifically in short styles in distylous common buckwheat (Fagopyrum esculentum). Heredity, 123(4), 492–502. https://doi.org/10.1038/ s41437-019-0227-x Thakur, P., Kumar, K., Ahmed, N., Chauhan, D., Rizvi, Q. U. E. H., Jan, S., et al. (2021). Effect of soaking and germination treatments on nutritional, anti-nutritional, and bioactive properties of amaranth (Amaranthus hypochondriacus L.), quinoa (Chenopodium quinoa L.), and
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Chapter 4
Brown Rice Sprouts: A Leading Functional Food Product Bharat Garg, Shikha Yashveer, Manjeet Singh, and Jyoti Duhan
Rice is considered a staple food in a number of different cultures, but it is especially common in South, East, and Southeast Asia (Mridha et al., 2022). The cereal grain is consumed the second most frequently after wheat. Figure 4.1 symbolise the pericarp and aleurone layers that make up the major part of the grain: the husk, the endosperm (white rice), the germ, and the bran. It is also the dominant contributor to carbohydrates in the diet. People are now drawn to meals with healthy and natural ingredients to help keep their immune systems strong and prevent heart disease (Meydani, 2000). The role of rice in a country’s diet and how it is processed vary from one country to the other (Fig. 4.2). Most people eat white rice, made by removing the husk as well as bran layers, which contain nutrients and chemicals that are good for our health (Patil & Khan, 2011; Watanabe et al., 2004). After harvest, to make rice easier to consume, the grain is frequently dried, milled, and packaged. The first stage in husk milling is removing the paddy to uncover the brown rice grain (BR) below. BR has an outer coating of bran that is brown in colour (Saleh et al., 2019). There are more bioactive components like γ-oryzanol, phenolic acids, Gamma-aminobutyric acid (GABA), flavonoids, γ-tocotrienol, and α-tocopherolin the entire BR grain (Pang et al., 2018). These components are found in very high concentrations in the rice bran layers covering the grain’s exterior (Sharif et al., 2014). On the other side, BR has a wrong texture B. Garg (*) · S. Yashveer Department of Molecular Biology, Biotechnology & Bioinformatics, College of Basic Sciences & Humanities, CCS Haryana Agricultural University, Hisar, Haryana, India M. Singh Department of Genetics & Plant Breeding, College of Agriculture, CCS Haryana Agricultural University, Hisar, Haryana, India J. Duhan Department of Biochemistry, College of Basic Sciences & Humanities, CCS Haryana Agricultural University, Hisar, Haryana, India © The Author(s), under exclusive license to Springer Nature Switzerland AG 2023 I. Majid et al. (eds.), Advances in Plant Sprouts, https://doi.org/10.1007/978-3-031-40916-5_4
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Fig. 4.1 Different parts of rice grain
Fig. 4.2 Rice category types
and is hard to eat. Due to these drawbacks, sprouted brown rice (SBR) was already made available (Cho & Lim, 2016). Also, BR must germinate (which makes SBR) to get the nutrients needed for good health. Germination makes the boiled BR kernels absorb more water and become softer, which makes them better to eat. People are interested in SBR because it is softer and tastes better than milled rice and BR. It’s also better for our health. Many studies have shown that taking GBR frequently helps prevent long term diseases like colorectal cancer (Shao & Bao, 2015). GBR also helps to prevent Alzheimer’s disease because it has more GABA than other foods. GBR is an adequate food because it is easily digestible and has more bioactive components than regular BR. The way to make GBR is straightforward. Depending on ambient temperature, the BR is immersed for one or two nights before germination. Because this procedure alters the BR’s minerals, the finished product is more delicious, healthier, and simpler to chew. It has been hypothesized that the GBR involved in improving brain function and lowering in the number of lipids in the blood.
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4.1 What Is SBR? “Germinated brown rice(GBR)” is another name for SBR. Germination is a process that starts when a dry seed takes in water and ends when the embryonic axis forms (Montemurro et al., 2019). Sprouts are grown from grains that have germinated. This is a simple process that usually does not need sunlight or soil. So, it does not depend on the season. It is essential in the summer and when it rains, when fresh veggies are harder to find (Bains et al., 2014; Cauchon et al., 2017). GBR fixes the problems with BR because it has a better quality, nutrition, and health benefits (Ren et al., 2020; Yodpitak et al., 2019). During the soaking process, the water is quickly absorbed. Several biochemical processes soften the texture, break down polymers, and speed up some phytochemical compounds’ manufacturing process and buildup (Cho & Lim, 2016; Yodpitak et al., 2019). Rice can be germinated by immersed it into lukewarm water (35–40 °C) over ten hours, draining it, and keeping it moist for 20–24 h. Every 3–4 h, the water should be changed to prevent microbial activity (which usually gives off a bad smell) and to keep the water temperature consistent during the soaking period. The result is a sprout about 1 mm long that grows from the BR grain. The grain gets the most nutrients (Patil & Khan, 2011).
4.1.1 Germination The metabolic changes of germination begin by turning on the enzymes that are still in the seed. As a result, numerous studies on the process of grain sprouting have investigated the changes in enzyme activity as well as the composition of the grain that occur as a direct result of germinating. Understanding the biosynthetic activity of germination is essential to controlling or improving the composition changes during germination. Germination usually starts when dry seeds take in water and meet the right specific environment to grow. In BR, sprouting is the process up to the extent in which the rice seed has a root system that is 3–5 mm long and pre-germination stage when the rice grain has a root system that is about 0.5–1 mm long and has started to grow (Watanabe et al., 2004). Even though there are differences between plant species in how they take in water and how their enzymes react, germination can be broken down into three stages: the first stage, known as the imbibition process; second stage is the stimulation of metabolism (respiration and the metabolism of carbohydrates), and final stage is the growth of roots and shoots, as well as the emergence from the husk (Figs. 4.3 and 4.4).
4.1.2 Parameters for Germination For the factory output of germinated rice with regular qualitative and quantitative characteristics, it is crucial to find the best conditions for the germination of BR. Both internal and external factors affect germination. Internal factors include
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Fig. 4.3 Germination stages
Fig. 4.4 Phases of germination. (Yodpitak et al., 2019)
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the cultivar of BR and how the brown rice is milled and stored. External factors include temperature, humidity, O2 or air, amount of light exposure, and acidity level. For BR seeds to grow, they need to be in contact with water. There are two ways to do this: soak the seeds or put them in water for a short time and then let them grow in the air. The germination rate was higher with atmospheric germination than just soaking in water, so the germinated rice had deeper and more extensive roots and sprouts (Lu et al., 2010). Capanzana and Buckle (1997) tested malting level, protein, thiamine, and amylase activity under different soaking and sprouting conditions. After soaking for 24 h at 25 °C, they determined that sprouting in the atmosphere at 30 °C for three days is ideal for BR. Long germination periods cause microbial activity and cause roots and sprouts to grow too much, making GBR often unfit for use as food. The growth of microorganisms and the development of sourflavours can be slowed down by rinsing the seeds often while soaking or germinating in the air. Microorganism growth could also be slowed by changing the ingredients in the cleaning solution, such as using chitosan solutions or electrolysed water (Lu et al., 2010). To produce GBR, you should also think about the germination percentage, microbial contamination, and physiological abnormalities in bio-functional components.
4.1.3 Bioactive Components in GBR Recent researches have revealed that sprouting can cause bioactive chemicals to build up in GBR seeds and sprouts. These compounds include γ-oryzanols, a lipophilic fraction, and the main substances in BR. Vitamin E, phenolics, and phytosterols are some of the other phyto-compounds in BR. Through the germination process, these bioactive components can be made or changed. So, GBR has higher levels of some of these bioactive substances than BR, which suggests that germination may make BR more bioactive. All structures are downloaded from Pubchem database. Here, we give a brief overview of the main bioactive components listed in Table 4.1 and the analytical methods used to find them in Table 4.2. 1. Antioxidants Antioxidants are molecules that guard the body’s tissue from detrimental superoxide anion and oxygen free radicles; they could be negative impact for a person’s metabolism if they are not stopped. Phytochemicals are chemicals found in plants that can help fight free radicals and have other health benefits. Phenolics are plant chemicals with hydroxyl groups from the phenyl ring together with an antioxidant effects (Van Hung, 2016). Most of these nutrients can be found in BR, but phenolic acids are the most prevalent (Gong et al., 2017). Phenolics are a phytochemical type with ring structures together with hydroxyl groups (Tan et al., 2017). Some good examples of phenolics are polyphenolic compounds, tannins, flavonoids, stilbenes and
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Table 4.1 Main bioactive components in GBR Class Family Antioxidants Phenolics
Flavonoids
Oil components
proanthocyanins and Anthocyanins Vitamins Phytosterols γ-Oryzanol
Others
DFs Amino acids
Compounds Syringic acid, ferulic acid (cis, trans), feruloylsucrose,Gallic acid, Trans-p- coumaric acid, vanillic acid, caffeic acid, sinapic acid, ellagic acid, protocatechuic acid, p-hydroxybenzoic acid Tricin, Flavones, quercetin, apigenin, kaempferol, myricetin cyanidin-3-O-galactoside, cyanidin-3-O-glucoside, epicatechin, catechin Tocopherols, B vitamins (B1, B3, B6), tocotrienols Stigmasterol, β-sitosterol, stigmastanol, δ7-avenasterol, campesterol Cycloartanylferulate, campesterylferulate, 24-methylene cycloartanylferulate Pectin, hemicelluloses and mucilage Alanine, aspartic acid, arginine, glutamic acid, glycine, cystine, histidine, leucine, lysine, isoleucine, serine, threonine, phenylalanine, tryptophan, valine, tyrosine
Table 4.2 Analytical methods Sr. No. 1
3
Method of analysis Microwave-assisted extraction (MAE) Ultrasound-assisted extraction (UAE) Fluorescent DCF
4
UV-visible spectroscopy
5 6
Amino acid auto analyser Reversed-phase HPLC method Gas chromatography HPLC method
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Enzymatic-gravimetric method
Bioactive compound Phenolic acids, Antioxidants
References Sato et al. (2004)
Phenolic acids, Antioxidants
Sato et al. (2004)
Flavonoids, Tocopherols and tocotrienols Anthocyanins, proanthocyanins and Phytic acid GABA γ-oryzanol
Srisawat et al. (2010) Perera et al. (2018)
Phytosterols Essential amino acid DFs
Cao et al. (2015) Xu and Godber (1999) Zubair et al. (2012) Amankwah et al. (2015) Tiansawang et al. (2016)
coumarins (Liu et al., 2007). Rice has three different kinds of phenolics: unbound, solublized, and linked. The linked form is the most common of the three (Adom et al., 2002). There are many phenolics in the germ and rice husk coatings (Adom et al., 2005). Because BR is not polished or milled, the phenolics in the germ and husk coatings stay in good shape. The small intestine is the best place to absorb free phenolics.
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In contrast, bound phenolics tend to stay in the intestines and are only removed through the intestinal wall, where they engage with the microflora to promote the ratio of Firmicutes to Bacteroidetes (Martínez et al., 2013). Monohydroxybenzoic acid and 4-coumaric acid make up the two major subgroups of phenolic acids (Liu, 2007). BR has a lot of phenolic compounds. The main one is trans-ferulic acid, a bound hydroxycinnamic acid (Gong et al., 2017). The most prevalent polyphenols in BR is trans 3-hydroxycinnamic acid, a hydroxycinnamic acid derivative that makes up about 98% of the bound form (Gong et al., 2017). The bound form of cis- ferulate, an analogue of trans-ferulic acid, is also present in large amounts (Gong et al., 2017). BR has three main soluble phenolic compounds: feruloylsucrose, sinapoyl sucrose, and ferulic acid (Fig. 4.5). BR is different from white rice because it still has germ and bran layers, which are full of nutrients like antioxidants (Cáceres et al., 2017). So, even though BR is healthier than white rice, it is eaten less often because of how it looks, how long it takes to cook, how much it costs, how little it is available and how few people know how healthy it is (Adebamowo et al., 2017). Aside from cooking, several methods, such as germination, have been emphasized to make BR taste better and make its nutrients more bioavailable. Germination changes the texture and makes it easier for the nutrients and phytochemicals to be used by the body (Tian et al., 2010; Patil & Khan, 2011). Also, these phenolic compounds play a big part in preventing some long-term illnesses, like sugar related disorders, heart diseases etc. (Hudson et al., 2000; Jung et al., 2007). Flavonoids are another type of antioxidant found in BR. Flavonoids has a 15-carbon skeleton that is made up of two hydroxyl rings together with heterocyclic ring. The phenolic hydroxyls in flavonoids (Flavanols) are what make them work as antioxidants. Most flavonoids in BR are flavones, and tricin (Fig. 4.6) is the most important flavonoid, making up more than a 75% of overall flavonoids in BR (Goufo et al., 2014). In low amounts, other flavonoids like apigenin, luteolin, isorhamnetin, quercetin, myricetin, and kaempferol (Goufo et al., 2014). 2. Gamma-aminobutyric acid It was identified as an important component of the vertebrate central nervous system (CNS), gamma-aminobutyric acid, also known as GABA (Fig. 4.7), has served as the primary inhibitory neurotransmitter in the mammalian cortex. It was studied in plants 50 years ago and is made by a chemical process called the “GABA shunt.” Even putting the seeds in water before they sprout may be enough to raise the GABA content. Because GABA cannot pass through the blood-brain barrier, it may help lower blood pressure in the vascular and autonomic nervous systems (Hayakawa et al., 2002). The antihypertensive effect of GABA may come from its effect on presynaptic GABAB receptors, which reduces the release of noradrenaline (Hayakawa et al., 2002). GABA-rich GBR fractions stopped the growth of some cancerous cells and boosted the immune system (Oh & Oh, 2003, 2004).
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Fig. 4.5 Structure of some essential phenolic acids in BR
3. Oil components Most of the biofunctional materials in BR and GBR are “lipophilic,” which means they like fat. The by-products of milling rice, especially the bran and embryo, have much oil. There is not enough research on how the rice bran oil (RBO) in GBR works in the body. However, some studies showed that germination did not change the number of oil components like phytosterol (Fig. 4.8), c-oryzanol, squalene, and vitamin
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Fig. 4.6 Structure of some important Flavonoids in BR Fig. 4.7 Gamma- Aminobutyric Acid (GABA)
E or made them a little bit more (Jayadeep & Malleshi, 2011). Kwak et al. (2013) looked at how the lipid components in BR changed during sprouting. They found that many lipid components in GBR oil did not change. Even though RBO has bioactive components, it has not been used very often because it is easy to oxidise. Lichtenstein, Ausman, and Carrasco found in 1994 that RBO lowers cholesterol more than other vegetable oils. RBO may have a high effect on lowering cholesterol because it has bioactive components that are not usually found in other plant oils. These include c-oryzanol, linoleic, tocotrienols, and phytosterols. 4. Gamma-oryzanol Gamma-oryzanol was made up of a group of ten or more compounds that were linked to triterpenes and ferulic acid by ester bonds (Fig. 4.9). The main compounds of c-oryzanol in GBR are cycloartenyl ferulate, 24-methylene cycloartenyl ferulate, and campestanyl ferulate, which make up about 90% of GBR c-oryzanol (Jayadeep & Malleshi, 2011). The main job of the ferulate part of c-oryzanol is to fight free
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Fig. 4.8 Structure of important phytosterols
Fig. 4.9 γ- Oryzanol
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radicals. Using linoleic acid as a model, a study found that the main components of c-oryzanol (24-methylenecycloartanyl ferulate, cycloartenylferulate, and campestanylferulate) stopped linoleic acid from being oxidised by ultraviolet light. However, their effect was not as strong as free ferulic acid and a-tocopherol (Xu & Godber, 2001). This antioxidant effect of c-oryzanol on cholesterol oxidation is probably linked to the fact that it lowers cholesterol. Researchers have shown that c-oryzanolcan lower cholesterol (Wilson et al., 2007). 5. Tocols Vitamin E (tocopherols), most important chain-breaking antioxidant that dissolves in lipids, also crucial for protecting the body’s nerve cells. The methyl groups on a chromanol ring are used to sort the tocopherols into different groups. Tocopherols are those whose chromanol rings have a saturated phytyl tail, while tocotrienols are those whose isoprenoid side chains are not saturated (Fig. 4.10). Germination changes the composition and amount of vitamin E, primarily independent of the type of rice. The most vitamin E is found in GBR in c-tocotrienol, followed by a-tocopherol, tocotrienol, and c-tocopherol (Jayadeep & Malleshi, 2011). The c-tocotrienol is also very good at stopping cancer cells from growing. Kannappan et al. looked into the effect of c-tocotrienol on the death of tumour cells (apoptosis) in 2010. In a way that depended on dose and time, Gamma-Tocotrienol could stop the activation of the STAT3 mechanism that is closely associated to tumour growth. They also found that c-tocotrienol turned off the proteins VEGF, Cyclin D1, and MMP9, which are needed for tumour cell growth, blood vessel growth, and movement. 6. Policosanols Long-chain aliphatic primary alcohols with 20–35 carbons are called policosanols. Some examples are 1-tetracosanol, 1-heptacosanol, 1-hexacosanol, 1-nonacosanol, 1-octacosanol, 1-tetratriacontanol, 1-triacontanol and 1-dotriacontanol. They are discovered in variety of plant parts, including seeds, fruits, leaves as well as nuts. They can be extracted from sugar cane, beeswax, sorghum, wheat, maize and BR.
Fig. 4.10 Tocol
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7. Other Oil Components BR has more bioactive oil components than c-oryzanol, policosanol and tocols. These substances are much less than GABA or c-oryzanol, but their biofunctions seem essential. Also, little is known about how germination changes its composition and functions. Plants have many phytosterols, but stigmasterol, campesterol, and b-sitosterolare the most common ones (Brufau et al., 2008). Plant sterols are more than 100 different phytosterols and more than 400 other triterpenes (Moreau et al., 2002). phytosterols and triterpenoids are both natural substances that come from hydroxylated polycyclic isopentenoids with a structure of 1,2-cyclopentanophenanthrene. 8. Dietary Fibers (DFs) DFs are carbohydrate polymers that can be eaten but are not digested by small intestine enzymes. They can be put into two groups: those that dissolve in water and those that do not. Soluble DFs includes pectin, mucilage, and some hemicelluloses. They thicken the stool in the intestines and reduce the blood sugar and cholesterol in the body. Insoluble fibers like cellulose are facilitate to keep the colon healthy by increasing the volume of faeces, helping the bowl move, and getting rid of toxic metabolites. It was said that the anti-diabetic effect of GBR came from the insoluble compound DFs (Seki et al., 2005). Using male Wistar strain rats fed a control diet (white rice), Seki et al. (2005) found that removing oil molecules and soluble components from destarched and macerated pre-GBR bran lowered post-prandial glucose levels and plasma insulin concentrations. A high fiber diet may lower a food’s glycemic index and the number of people with Type 2 diabetes. DFs are found in many foods, but if we eat BR and GBR as our main foods, we may get enough of this health-promoting substance.
4.1.4 Germination Effects on BR Nutritional Content Dry seed’s metabolic activity rise as soon as it gets watery during soaking. During germination, different parts of the grain undergo many changes at biochemical level. During sprouting, only water and oxygen are taken in by the seed. Because no outside nutrients are added, most of the good nutritional changes come from breaking down complex compounds into simpler ones and turning them into essential parts, as well as breaking down parts that are bad for the seed’s health (Chavan & Kadam, 1989). 1. Changes in Starch and Sugar Reduction as a Result of Germination Starch is the primary ingredient in rice kernel (Zhou et al., 2002). The endosperm is where this polysaccharide is stored most of the time. During germination, it is broken down into sugars that the seedling can use (Ayernor & Ocloo, 2007). The starch breakdown is a complicated biochemical pathway controlled by hormonal and
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metabolic actions (Perata et al., 1997). For the metabolic changes in dry seeds to happen, they need enough energy. Most of this energy comes from the breakdown of residual starch, which makes up 90% of BR. After germination, the total amount of starch, amylose, and amylopectin in BR slowly decreases. The primary way that GBR loses starch is by reducing amylose (Zheng et al., 2006). Alpha-amylase may have been created during protein synthesis at the seedling stage rather than already being present in the seeds, even though a minor amount of alpha-amylase is discovered in dry seeds (Moongngarm & Saetung, 2010). Furthermore, α-amylase, the enzyme responsible for the conversion of starch into maltose units, is primarily found in BR kernels and starts to work when imbibition begins (Palmiano & Juliano, 1972). Because of the actions of amylolytic enzymes, germination causes the quantity of amylose in BR to go down. Jiamyangyuen and Ooraikul (2008) looked at how germination affected the nutritional and sensory characteristics of BR. They found that germination made cooked GBR more cohesive and soft but made it less sticky and bland. Such things are probably caused by the way amylolytic enzymes break down amylose during germination and by the cracks that form in rice kernels throughout sprouting and drying. Moongngarm (2010) said that BR flour became much harder to make a paste with as germination time went up. This might have happened because the amylases and proteases turned on during germination broke down the starch and proteins connected to starch granules. 2. Variations in GABA Concentration The amino acid known as GABA is one that is absent from proteins. It is made when glutamate reacts with glutamate decarboxylase (Ding et al., 2019). The difference between the ungerminated BR samples and the germinated samples were significant (P .05). The most GABA was found in GBR samples that had been germinating for 36 h (P .05). From 0 h to 36 h, the germination process made 1.5 times as much GABA as before (He et al., 2022). BR already has glutamate in it, which is why it can make GABA levels rise when it germinates. Glutamate decarboxylase is triggered during germination to decarboxylate glutamate and generates GABA. BR’s GABA content rises due to this process (Diez-Gutiérrez et al., 2020). 3. Changes in Protein and Amino Acids Several researchers have looked at what happens to the storage proteins in BR when it germinates. Zheng et al. (2007) discovered that sprouting enhanced the amount of albumin and gluten proteins in BR and lowered the amount of globulin and gliadin proteins. This made the proteins more bioavailable. Chen et al. (2003) found that the amount of soluble protein went up for the first six days of germination but then went down. Intact rice grains were immersed in bottled water for half day at 30 °C and allowed to sprout in the dark for five days by Veluppillai et al. (2009). The results demonstrated that the amount of total protein dropped significantly during germination. In contrast, the amount of soluble protein dropped until the second day and rose until the fifth day. During germination, the quantities of majority of the amino acids went up a lot. Veluppillai et al. (2009) discovered that the number of amino acids (free) in BR
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grew from 1.96 mg/g to 3.69 mg/g of dry matter during germination, which can take 4–5 days. This means that BR’s taste also improves after it germinates. Saikusa et al. (1994) found that after 4 h of soaking, the levels of amino acids in different rice husk fractions rise significantly. The necessary amino acid profile of GBR is more similar to the FAO/WHO model than that of BR, according to Zheng et al. (2007). Serine, asparagine, aspartic acid, and glutamic acid during BR germination decreased. However, all other amino acids increased, according to Komatsuzaki et al. a strong correlation was found between the germination circumstances and variations in amino acid levels, with thermal treatment helping to rise the content of amino acids in GBR. 4. Changes in Lipids Few studies have been done on how germination changes the lipids in BR. Shu et al. (2008) discovered that oleic and palmitoleic acid concentrations went up at the beginning of germination but went down quickly after 72 h. Their results are a bit different. Up to 72 h, the amount of free fatty acids might have gone up because lipases produced during germination broke down lipids or because starch was broken down and changed. After 72 h, the amount of free fatty acids might have gone down quickly because they were being used more. 5. Changes in Phenolic Acids Compared to BR, the amount of feruloylsucrose and sinapoyl sucrose in GBR has dropped by about 70%. In contrast, the amount of free ferulic acid has gone up (Tian et al., 2004), But GBR has less soluble phenolic compounds than BR (Tian et al., 2004). Aside from that, the quantity of sinapinic acid in seedling rice is 10 times greater than it is in non-germinated rice (Tian et al., 2004) 6. Changes in γ-Oryzanol Researchers have looked at how the germination process affects the quantity of γ-oryzanol. It was studied that the amount of γ-oryzanol in germinated as well as non-germinated rice is almost the same. It was also discovered to be 21 times more abundant in BR than in white (Miura et al., 2006). GBR has a slightly greater concentration of γ-oryzanol (105%) than BR according to Ohtsubo et al. (2005). BR cultivars differ in how the germination process affects the quantity of γ-oryzanol they produce according to Kiing and colleagues (2009).
4.1.5 Effect of Soaking Condition on Germination Due to the biological perks of germinated over non germinated rice; much research has been conducted to enhance the sprouting procedure. Wet condition is essential for sprouting, and it can be done in two ways: by completely soaking the seeds in water or letting some of the seeds sprout (Cho & Lim, 2016). Many studies have shown that these two ways can cause germ to grow, and change the physical characteristics and biologically active components of germinated rice (Zhang et al., 2014;
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Cáceres et al., 2017). Soaking is, hence, one of the most critical steps in making GBR. During manual soaking, the water should be changed every so often to stop microorganisms that cause food to go wrong from starting an uncontrolled fermentation process (Pinkaew et al., 2016). When done on a larger scale, this process is less effective and creates a lot of waste water, so it is not considered suitable for the environment. So, a plan is needed to reduce the amount of wastewater made and optimize the soaking process during GBR production. One way that BR could be soaked is through the filtration process. The polymeric membrane is starting to look like a good choice for filtration. The idea that a reactor with a microfiltration membrane could be used to make GBR is interesting. Based on what was said above, the goal of this investigation was to use the membrane reactor to make GBR and to find out how different sprouting methods affect GABA and other bioactive compounds (Munarko et al., 2021). Length of a Sprout GBR sprouts get longer when they take a long time to grow from seeds. The RFS (Reactor complete soaking) method and the MFS (Manual full soaking) method had the highest and lowest growth rates, respectively. GABA and Glutamic Acid Proportion In non-germinated rice, there are no traces of GABA content. Following 24 h of immersion, GABA was found both in the reactor and by hand. Using RAG, the amount of GABA in the seed grew significantly and reached its highest level after 60 and 72 h of germination. In contrast, the MFS treatment caused GBR GABA levels to rise at the slowest rate. Antioxidant Capacity TPC of BR before germination decreased after 24 h of soaking in both the reactor and by hand. The phenolic acids in BR may have been partially washed away into the imbibation liquid (Cáceres et al., 2017; Cho & Lim, 2018), which could explain why TPC went down in the early stages of soaking. γ-oryzanolcontent γ-oryzanol amount changed slightly when it started to grow. At the end of the initial 24 h of soaking, the amount of γ-oryzanol in the seedling rice soaked in the reaction chamber had not changed much. In sprouted rice made with the RAG, RFS, and MAG methods, the total content of γ-oryzanol decreased slightly after 24 h.
4.2 The Impact of Calcium Chloride (CaCl2) Treatment on the Accumulation of Bioactive Components and the GBR’s Ability to Fight Free Radicals (Choe et al., 2021) Soluble calcium ions may be a crucial part of making BR that has germinated and is rich in bioactive compounds. In this case, it is essential to know if adding abiotic elicitors like calcium chloride (CaCl2) during the sprouting of BR could enhance its ability to fight free radicals and cause it to make more bioactive compounds.
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Fig. 4.11 Effect of calcium chloride
After the germination treatment, the proportion of total phenols, GABA, and total flavonoids in the BR went up significantly (Fig. 4.11). Elicitation with CaCl2 of GBR changed the content of GABA, total phenols and flavonoids in a big way. GBR treated with different amounts of CaCl2 grew a lot. The number of flavonoids in GBR treated with 200 mM CaCl2 was also 132% higher than in GBR used as a control. Under 200 mM CaCl2, the most GABA was found in BR that had germinated. As signaling molecules in physiological modulatory networks, keep the structure and function of cellular membranes intact and control ion flow and selectivity in relation to outside stimulation (Hepler, 2005; Sheen, 1996). CaCl2 is essential for plants to recover from damage and start repairing their cells. It controls cellular processes like antioxidant enzyme activity and lipid peroxidation (Khan et al., 2010; Tan et al., 2011). Adding CaCl2 to BR during germination greatly impacted the rise in PAL activity but had not effect on TAL expression. When CaCl2 is added to grain sprouts from the outside, the total amount of calcium and calmodulin increases significantly. This may alter genes and protein expression and PAL enzyme function, associated in polyphenol biosynthesis, and so impact the overall phenolic content (Ma et al., 2019).
4.2.1 Plasma Treatment to Improve the Germination of BR Plasma is a sterile, ionizing gas and the fourth most energetic form of energy (Moreau et al., 2008). This procedure can modify how seeds absorb water, which can help them sprout faster and produce more crops (Randeniya & de Groot, 2015). Also, seeds can be kept for extended periods after being treated with cold plasma. Germination test is conducted by taking 100 grains of both BR and plasma- treated BR, after that put on filter in 12 cm cell culture dishes and 10 mL of bottled water was added. During the tests, every 4 h, the bottled water was changed. The germination rate was calculated by dividing 100 times of germinated seeds by the
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cumulative number of seeds, and the morphological characteristics (overall root length and plant height) were measured (Yodpitak et al., 2019). The dielectric barriers were made out of ceramic. In all the tests, the distance between two diodes was kept the same (0.5 cm). A radio-frequency power source was hooked up to the DBD. The rice crops were uniformly spaced apart between the two electrodes on the aluminium sample tray. At room temperature and atmospheric pressure, plasma was used to test the samples.
4.2.2 Health Benefits of GBR People worldwide have become more interested in natural and healthy foods in the past few years. This has led to food science to study the link among both eating habits and health, and a fast-growing nutraceutical market (Viuda-Martos et al., 2010). Researchers are becoming increasingly interested in the health benefits of GBR, and some have found a link between GBR foods and to keep certain diseases from happening. GBR has been found to have many healing properties, like reducing cholesterol and blood pressure, and also decrease the likelihood of diabetes, coronary heart disease (CVD), cancer, and Vascular dementia. It is therefore considered a functional food. 4.2.2.1 Antihyperlipidemic Effect Hyperlipidemia, also called dyslipidemia, is a major cause for cardiovascular diseases. It is caused by an imbalance between the energy we take in and how much we use, which can be caused by insufficient exercise (Braunwald, 1997; Nelson, 2013). In recent years, as health care costs have increased, hypocholesterolemic, anti- diabetic, and anti-obesity functional foods have become very popular worldwide (Mermel, 2004; Siró et al., 2008; Ejtahed et al., 2012; Albarracín et al., 2016). Miura et al. (2006) discovered that GBR and BR slowed the growth of hepatomas- caused high cholesterol by speeding up the breakdown of cholesterol. Their tests also showed that GBR was more effective than BR at restoring HDL cholesterol levels that had dropped because of a hepatoma. BR that has germinated may be good at lowering cholesterol because it has a lot of bioactive parts, such as GABA, γ-oryzanol, DFs, or other antioxidants. It was said that plasma cholesterol levels in hamsters fed a diet with cholesterol were much less in those fed a rice bran diet than in those fed a cellulose diet (Kahlon et al., 1992). By stopping cholesterol from getting into the bloodstream, γ-oryzanol was said to lower plasma cholesterol in hamsters whose diets contained cholesterol (Rong et al., 1997). More in-depth research is needed to determine what chemicals in GBR make it work against high cholesterol and the exact mechanism behind these effects.
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4.2.2.2 Antihypertensive Effect Hypertension is a lifestyle disease. It causes heart attacks and kidney problems. Hypertension is treated using calcium channel blockers, ACE inhibitors, vasodilators, cardioselective blockers, and receptor agonist medicines (Perez & Musini, 2008). It has been shown that GBR can help to lower blood pressure (Yuji et al., 2004; Ebizuka et al., 2007, 2009). GBR may be able to lower blood pressure because of how its many different parts work together. These parts include GABA, DFs, and ferulic acid. Ferulic acid, whose amount gradually increased after seeding, helped lower the blood pressure of rats with diabetes caused by streptozocin and rats whose blood pressure was high on its own (Ardiansyah et al., 2008). Based on this, GBR, which has higherphytochemical constituents than normal rice, should be a good option as the leading food or functional food for lowering blood pressure. 4.2.2.3 Anti-cancerous Effect Cancer remains a leading cause of death worldwide. Chemotherapeutics is a technique for avoiding cancer by eating chemical compounds that make it less likely that cancer will form (Bae et al., 2002; Steele, 2003). Some studies have been written about how certain parts of rice might help prevent cancer. Arabinoxylan hemicellulose is inedible rice feed fiber compound that makes it easier for rodents to get rid of carcinogenic xenobiotics through their intestines (Takenaka & Takahashi, 1991). It also stopped Dimethylhydrazine Hydrazomethane from causing cancer in rodents’ intestines, which was caused by hemicelluloses (Aoe et al., 1993). Based on these results, replacing white rice with BR and rice bran may be suitable for preventing cancer. Because of this, GBR, which keeps most of the bioactive parts in the rice kernal and germ and also has extra bioactive parts than non-GBR, might be a better option for a cancer-prevention staple food. Also, scientists have looked into the effects of GBR on the growth as well as death of tumor cells (Oh & Oh, 2003; Tian et al., 2004). These results also show that GBR may be suitable for preventing cancer if it is a staple food. 4.2.2.4 Anti-diabetic Effect Lifestyle diseases like diabetes have become a leading public health issue, and much evidence shows a link between diabetes and many kinds of cancer (Chen et al., 2015; Li et al., 2017; Wang et al., 2020). Because of this, it is imperative to develop different kinds of functional foods to help with this problem (Binh et al., 2020). Researchers have shown that GBR is suitable for people’s blood sugar levels when fasting, after eating, and when they respond to insulin. It was found that both non-diabetic and diabetic patient whose diets included GBR had lower blood sugar
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levels after eating than those whose diets included white rice (Ito, 2005; Ito et al., 2005). Concerning the outcomes of eating GBR over a long period, one study looked at the sugar and lipid levels of people with low fasting glucose or diabetes type II who ate either white rice or GBR for six weeks with a 2-week break in between (Hsu et al., 2008). It was said that diabetic patients who ate a lot of germinated rice might have lower blood sugar levels and less risk of diabetic vascular complications.
4.2.3 Cardiovascular Disease Prevention Myocardial ischemia is still one of the main reasons people worldwide get heart failure (Ahluwalia et al., 2011). Reperfusion is the main way to get blood to start flowing again to the heart. However, ischaemia-reperfusion causes damage to heart tissue, problems with how the heart works electrically, and heart failure (Lisa et al., 2006). Researchers have found that too much generation of reactive oxygen species (ROS) after cardiac surgery triggers oxidative stress, which damages cardiac myocytes (Clark et al., 2007). So, an antioxidant agent from the outside may be needed to minimize and stop the damage caused by oxidative stress during myocardial reperfusion. Several investigations (Yin et al., 2013; Estes & Kerivan, 2014; Yu et al., 2015) looked into the antioxidant substances that could be used as a possible treatment for cardiac reperfusion injury. BR has many antioxidants in it from the way it grows. Components of BR like γ-oryzanol, GABA, ferulic acid, and phytic acid are a better alternative to treat chronic diseases and improve health (Kozuka et al., 2017). Petchdee et al. reported in 2020 that GBR helped ischemic reperfusion injury by making apoptosis less likely. GBR may help protect against cell proliferation and apoptosis and may also help prevent heart failure caused by myocardial ischemia.
4.2.4 Alzheimer’s Disease Prevention Alzheimer’s disease is one of the prevalent reason older people lose their ability to think and remember things over time. Theβ-amyloid (A) peptide found in senile plaques causes’ brain impairment, as shown by rodents’ neurotoxicity and problems with learning and memory (Nitta et al., 1994). Mamiya et al. (2004) examined the role of GBR on mice with cognitive and memory problems caused by β-amyloid protein. From what they saw, it seems that GBR enhances spatial learning. Especially, it was thought that GBR might protect against Alzheimer’s disease, which is linked to a protein called peptideA. Nevertheless, this conclusion needs to be checked by doing more experiments.
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There may be more than one way GBR helps people learn and remember things. BR germinated has a lot of GABA, an essential part of memory. It is well known that GABA is a major neurotransmitter that slows down nerve impulses in animals (Mody et al., 1994).
4.2.5 Functional Food Developed from GBR As more people want healthy, high-quality foods, the food organization and the scientific community have developed novel nutritional foods. Sprouts are becoming more popular because they are healthy and have bioactive compounds that protect against long-term illness (McRae, 2017). It is hard for the food sector to produce new items that meet people’s needs for organic, gluten-free, and vegan foods. One crucial example is ready-to-eat yoghurt formulations. GBR could be used to make ready-to-eat yoghurt-like products. This could boost the number of fermented non-dairy alternatives on the market, which have been scarce in western societies. The brewing process using lactic acid bacteria (LAB) may improve the health benefits of GBR because LAB is like cell factories that make nutrients and bioactive that makes cereals work better (Waters et al., 2015). Fermentation slightly changed the proximal ratios of BR active ingredients and increased the bioactivity, consistency index, ACE-inhibiting interaction, and saturation. The FGBR96 product that looked like yoghurt had the best biological and technological properties and was the most popular overall. Vegans and those who wish to consume less animal protein may consider F-GBR formulations to be natural, healthful food options.
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Chapter 5
Amaranth & Quinoa Sprouts Anamika Sharma, Masud Alam, Kirty Pant, and Vikas Nanda
5.1 Introduction In recent years, due to various environmental and health factors, people are more distressed about their changing lifestyle, and therefore, it becomes mandatory to maintain a health quotient. For this reason, many people have switched to functional and nutraceutical foods. Hence, sprouts of various grains are becoming popular day by day as they help in the enhancement of human health and lowers nutrient deficiencies. Further, this change in lifestyle is held responsible for various diseases and among such ailments is a celiac disease caused due to the consumption of gluten protein (i.e. from cereals like wheat, rye & barley etc.) in their diet. To overcome celiac disease, various other cereal crops, except wheat, rice etc., came into existence a long time ago, possessing similar nutritional & physical appearance but varying in the fact that they are gluten-free and hence, referred to as pseudo-cereals (Iftikhar & Khan, 2019). Pseudo-cereals are the dicotyledonous plant species whose seeds are used for flour preparation for pasta making and other different bakery goods with high nutritional value. Most widely consumed pseudo cereals are Quinoa, Sorghum, Amaranth, Buckwheat & Chia etc. (Baraniak & Kania- Dobrowolska, 2022). Moreover, sprouts of these pseudocereals can help patients to recover from illnesses because of their multiple health-promoting properties. In this chapter, we discussed amaranth and quinoa sprouts, their nutritional and chemical composition, changes that occurred during the germination process and their health benefits.
A. Sharma · M. Alam · K. Pant (*) · V. Nanda Department of Food Engineering and Technology, Sant Longwal Institute of Engineering and Technology, Longowal, PB, India © The Author(s), under exclusive license to Springer Nature Switzerland AG 2023 I. Majid et al. (eds.), Advances in Plant Sprouts, https://doi.org/10.1007/978-3-031-40916-5_5
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5.2 Amaranth Amaranth is associated with the Amaranthaceae family and the dicotyledonous genus Amaranthus L. which is well known from the Mesoamerican culture that flourished between 1300 and 1572 AD. The term Amaranthus emanated from the Greek name amarantos which denotes “one that does not wither” (Thapa & Blair, 2018). Later, in the sixteenth to seventeenth century, it widely spread as a vegetable, cereal, weed, or crop. Along with its use as a food, amaranth is a sacred plant which is worshipped in various ritualistic and religious ceremonies throughout the world, but now it serves as a significant grain and vegetable crop in countries like India, Nepal, Africa, China and Pakistan (Baraniak & Kania-Dobrowolska, 2022; Robinson, 1986). However, amaranth is a common crop in Peru, Mexico and Bolivia, but the largest producer of amaranth is China. Amaranth can be found as a grain crop, vegetable crop, weed and ornamental crop. Around 70 species are cultivated as annual plants, out of which the most widely grown amaranth grain species are Amaranthus hypochondriacus L., Amaranthus cruentus (L.) Thell. and Amaranthus caudatus L. On the other side leafy vegetable amaranth species grown are Amaranthus dubius, tricolor, hybridus. Other amaranth weed species include A. albus L. (tumbleweed), A. retroflexus L. (redroot pigweed), A. spinosus L. (spiny amaranth), A. palmeri (Palmer amaranth) (Robinson, 1986; Thapa & Blair, 2018). Amaranth is an annual and abiotic stress-tolerant crop (drought, high salinity, heat, heavy metals) that grows rapidly with an average height of 4–6.5 feet and leaf size ranging from 5 to 15 cm in a variety of colours (light green, dark green, reddish, pink, and black). Leaves of amaranth are elliptical in shape, without stipules and are petiolate opposite or alternating with inflorescence of 1.6–2.9 feet in length and red, purple or yellow in colour. The fruits of the amaranth plant are dehiscent with one seed inside it. On ripening grains, colour lighten and germinate rapidly in humid environmental conditions (Mbwambo et al., 2013). To maximise benefits, small farmers started growing amaranth both for harvesting leaves and grains. This potential crop uses C4 pathway for photosynthesis like maize and sugarcane, which convert atmospheric carbon into sugar per unit of water lost as compared with other plants that follow the C3 pathway of photosynthesis (Rice, Wheat, and Potato) (Emmanuel & Babalola, 2022) Amaranth gained its attention when it was elected as a major potential crop among 36 world’s utmost favourable crops in a research conducted by US national Academy of Sciences under the project “Underexploited Tropical Plants with Promising Economic Value”. Ever since then research on amaranth seeds progressively intensified owing to its high nutritional and agrarian potential (Soriano-García & Aguirre-Díaz, 2019). The seeds of amaranth are a rich source of micronutrients (vitamin and minerals) and macronutrients namely fats, dietary fiber and proteins. Amaranth seed protein is in plentiful amount ranging from 13% to 19% with balanced concentration of essential amino acids. In addition, it contains high level of oil, lipid, starch and antioxidants. Due to its gluten free properties, it can be used as an alternative to gluten products for those suffering from celiac disease. Sprouts being popular these days offer great advantage to
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pseudo-cereal crops for sprouting as they improve nutritional and functional properties. Pseudo-cereals-based sprouts can be used for distant food preparatory as a breakfast food, pasta, soups, salads etc.
5.2.1 Amaranth Seeds and Sprouts: An Overview Amaranth is regarded as an underutilized crop worldwide and it came into light, when it was considered as a storehouse of nutrition. Since then, number of researches has been conducted to know its potential as vegetable and grain crop. Both the grain and vegetable crop are highly nutritious with protein, fat, energy, minerals (Calcium, Magnesium, Phosphorous), vitamins (majorly Vitamin A and C), fiber, and bioactive compounds but slight changes may occur depending on the type of species (Table 5.1). Amaranth grain is highly proteinaceous and of high quality in nature with an average content of 13–19%as compared to other cereal crops with high content of lysine (5%), which is a limiting amino acid in other cereals like wheat, rice, and maize. Also, protein of amaranth is rich in Sulphur-containing amino acid (4.4%), i.e. methionine and cysteine, which are generally present in pulses. Valine, Leucine, Table 5.1 Nutritional composition of Amaranth species per 100 g (USDA, 2011)
Chemical composition Fat (g) Protein (g) Energy (kcal) Fiber (g) Ca (mg) Mg (mg) P (mg) Fe (mg) Carotene (μg) Thiamine (mg) Niacin (mg) Riboflavin (mg) Folate (μg) Ascorbic acid (mg) Sugar (g) Carbohydrate (g) Starch (g) Saturated fatty acids (g) MUFA (g) PUFA (g) Phytosterols (mg)
Amaranth seeds (per 100 g) 7.02 13.56 371 6.7 159 248 557 7.61 5716 0.116 0.923 0.200 82.0 4.2 1.69 65.25 57.27 1.4 1.6 2.7 24
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Amino acid (gram) Arginine Alanine Tryptophan Threonine Isoleucine Serine Leucine Lysine Methionine Phenylalanine Glycine Proline Tyrosine Valine Histidine Glutamic acid
Value per 100 gram 1.06 0.79 0.181 0.55 0.58 1.14 0.87 0.74 0.22 0.54 1.63 0.69 0.32 0.67 0.389 2.25
Threonine and Isoleucine are also present in amaranth but quite smaller in amount. The Amount of protein in amaranth makes it a protein complement as recommended by the FAO/WHO, helping in building of the immune system. A considerable amount of starch polysaccharide is also present in the perisperm of grains with an average concentration of 50–60% of the total. Also, sugar is higher compared to other cereal crop like maize, rice and wheat (Table 5.2). Apart protein, amaranth has a substantial amount of oil i.e. 6–10% in comparison to other cereals out of which 77% is polyunsaturated fatty acid present in germ of the grain and contains saturated fatty acid and unsaturated fatty acid in the ratio of 1:3. These unsaturated hydrocarbons present in amaranth possess antioxidant, hypocholestrolemic and anticarcinogenic properties which help get rid of serious health issues. However, the concentration of fatty acid in oil vary with the type of amaranth but on general basis it contains 34% oleic acid, 33% linoleic acid, 19% palmitic acid, 9% docosaenoic acid, 3.4% stearic acid. It is also high in bioactive compounds like 10% of phospholipids, nearly about 8% of squalene, 2% tocopherols, and phytosterols nearly to 2% (Alegbejo, 2013). Amaranth is also a good source of vitamins like provitamin A, vitamin C (ascorbic acid), Niacin, Folate etc. and minerals like calcium, phosphorous and magnesium in an expressive concentration. On germination, these amaranth seed can improve digestible and nutritional properties by inactivation of anti-nutritional factors. Further, the process of germination enhances the corporeal characteristics of grain through the production of peptides via protein mobilization. Nutritional value of germinated amaranth seeds possess higher moisture content of 80–90% at the end of 72 h of germination,
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however protein of amaranth sprouts remains almost in similar concentration of about 13–17% and fiber content reduced during the germination process as shown by (De Ruiz & Bressani, 1990; Maurya & Arya, 2018) for the species A. caudatus. On contrary (Corzo-Ríos et al., 2021) found that fiber content increase with increase in germination time, which in turn is related to a decrease in time for gastric emptying as it reduces the intestinal transit time and therefore maintains the digestive regularities. Also, fat content depleted on germination of upto 72 h and the possible reason for this enormous destruction of fat could be because of increased lipolytic activity during the germination process which leads to the hydrolyses of fat components and further provides vigor for the development of the seedling (Paśko et al., 2009). Starch content on germination reduces by the action of α-amylase and β-amylase as given by (Balasubramanian & Sadasivam, 1989). However, due to the hydrolysis of starch by amylolytic enzyme, level of reducing as well as non-reducing sugars increases on germination for 24–48 h (Balasubramanian & Sadasivam, 1989; Kanensi et al., 2013). Amino acid content decrease with increase in germination time i.e. lysine as well as sulphur containing amino acid decrease (methionine and cysteine) due to structural modification in protein fractions. Besides this mineral composition improves on 24 h of germination which comprises of magnesium to reach at a level of 6221 mg/kg, Na levels increase to around 850 mg/kg which is much below the recommended daily intake of Na (Kanensi et al., 2013).
5.2.2 Germination of Amaranth Seed The process of sprouting a seed follows traditional household methods but also process of sprouting emerged as a controlled process by the manufacturer for getting an optimized product. Although sprouting is an intricate practice, but understanding the science behind, is also that much important in order to get a reliable product. Germination involves two main events that marks the initiation through the uptake of water and termination after extrusion of radicle from the seed coat. For sprouting, generally, two step procedure is followed steeping and germination. First phase of Steeping and imbibition, is a process in which grains are soaked in water till an ambient moisture content or hydration of cell content is achieved. As soon as the required moisture content is reached, extra water is removed and seeds are allowed for germination. Second phase involves great metabolic reactivation with slight uptake of water. Third phase of germination to proceed further requires aerobic conditions with constant temperature, airflow and high uptake of water to safeguard the undeviating germination rate. During germination, mobilization of storage reserves occur which is responsible for the physical and biochemical events which include: seed cover weakens, activation of metabolic activity, initiation in gene transcription, slackening of embryo cell wall, biogenesis of organelles, and most importantly changes in nutritional and chemical composition of edible product (Benincasa et al., 2019). When the process of germination completes, germinating seeds develops into sprout with a clear indication of radicle protruding out from the seed coat.
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Developed sprouts can be further processed into frozen sprouts or sprouts flour. Frozen sprouts are directly packed from the wet mesh material to frozen sprouts after freezing and for flour, sprouts are dried in kiln or in a dryer for the removal of water to 10–14% of moisture content (Finnie et al., 2019).
5.2.3 Changes in Chemical Composition During Germination 5.2.3.1 Carbohydrates Carbohydrates serves as the main storage reservoir present in kernels of almost every cereals and pseudocereals. Although starch being a principal form of carbohydrates storage reserve, few soluble sugars are also considered as carbohydrate storage. Sugars including sucrose and raffinose from are also identified in many of the seed plants (Białecka & Kępczyński, 2007). As discussed earlier, carbohydrate in the form of starch decreases with germination time; however amylase activity in ungerminated grain increase during the initial phase of the development. α-amylase and β-amylase activity during germination shows major change. α-amylase activity is highest in soaked grains which gradually falls with 96 h of germination. On contrary, β-amylase activity showed a rise between 48 and 72 h of germination and then falls off due to the fact that β-amylase is not able to solubilise native starch grains and act only on grains that were solubilised previously by enzyme α-amylase. Overall action indicates that content of starch diminishes with the rise in germination period as a consequence of hydrolysis. For this reason that hydrolysed starch gets converted into sucrose, sugar content (mostly in the form non-reducing sugars) enormously dropped after 48 h of germination which will be used up in later stages by the sprouts for the embryo growth (Balasubramanian & Sadasivam, 1989). Dietary fibre reduced significantly with the germination as discussed by (De Ruiz & Bressani, 1990; Gamel et al., 2006). Reports on reduction in dietary fibre concluded that germination of soybean also marked a decrease in fibre content due to α–L arabinofuranosidase activity. 5.2.3.2 Fats Fat as one of the main energy storage reserves is found in plenty in pseudocereals in the form of triacylglycerol. On the onset of germination, triacylglycerol content starts depleting by the action of lipases and gets degraded into glycerol and free fatty acids to endorse the growth of the seed. These degradation of free fatty acid with glyoxylic acid cycle and β-oxidation later gets transformed into sugars by the process of gluconeogenesis and ultimately providing required energy for the growth activities to carry further in the germinated seed (Liu et al., 2022; Thakur et al., 2021).
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5.2.3.3 Proteins Amaranth seed contains globulins and albumins protein stored in the vacuoles of the embryo and outside the protein bodies respectively. During the process of germination, protein mobilization takes place which involves the breakdown of vicilin globulin protein present in the storage vacuoles to hydrolyse initially with the action of protease and results in the liberation of amino acids while legumins are hydrolysed more slowly. Amino acid released after the carbon skeleton oxidation either provide energy or synthesise newer protein with small molecular peptides. Germination generally increases the protein content but on contrary several studies demonstrated that protein content decreases through the process of germination and the possible reason for this type of chemical change is the catabolism of protein. Also, reduction in protein after kilning of germinated grain was observed and concluded that catabolism of protein results into the production of sucrose which is bound with amino acid and protein of embryo to the radicle. Radicle undergoes the deculming step during kilning which degrades or removes the radicle and ultimately decreasing the protein content. Therefore, balance must be maintained between degradation of protein and biosynthesis of protein during seed germination (Aphalo et al., 2009; Thakur et al., 2021). 5.2.3.4 Vitamins and Minerals Processing of foods mainly serves the purpose of maintaining the nutritional value for a larger duration. Germination is one processing technique that increase the bioavailabilty of the minerals and improves the nutritional value. Amaranth seeds comprises of anti-nutritional factors like phytic acid hindering the bioavailability of minerals by binding with them. Minerals being an essential component for human health because of their properties of promoting mental and physical wellbeing of the person by developing the blood muscles, body tissues, nerve cells, teeth and bones (Bhinder et al., 2021; Nkhata et al., 2018) which makes it necessary to increase its bioavailaibilty. Therefore, germination makes this possible by phytase activation, hydrolysing phytic acid to inositol and phosphoric acid. and eventually intensifying the bioavailabilty. Research findings concluded that on germination of amaranth seed content of calcium and zinc increased while other minerals were not much affected by germination, due to reduction in anti-nutrients like phytate and tannins (Gamel et al., 2006). On the other hand, vitamins are also found to increase with the germination process. According to De Ruiz and Bressani (1990)) concentration of thiamine, folic acid, biotin, riboflavin, niacin, ascorbic acid increased gradually with increase in the germination period. The possible cause for this increase in vitamin content is metabolic activity of the amaranth seed grain. The increase of ascorbic acid during germination is due to starch hydrolysis by diastases and amylases (enzymatic hydrolysis) which further improves the glucose availability and acts as a precursor for the biosynthesis of ascorbic acid. On contrary, decreased ascorbic acid content was observed in two different varieties of
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amaranthus seed which may be due to the drying of germinated seed after germination. Niacin, niacinamide and riboflavin increased after germination and drying at 30 degree Celsius (Gamel et al., 2006).
5.2.4 Anti-nutritional Factors in Germinated Amaranth Seed Amaranth seeds contain various anti-nutritional factors which include phytic acid, tannins, lectins, saponins, cyanogenic glycosides and trypsin inhibitors. Report findings shows that species amaranthus hypochondriacus exhibit higher amount of trypsin inhibitors, lectins and decreased concentration of saponins and tannins. These factors could be slowed down with the use of thermal processing or germination. Germination decreased the trypsin inhibitors and the possible reason for such a reduction could be related with the increase of proteolytic enzyme activity which in turn reduces the content of trypsin inhibitors which promote the seed growth. According to the study, saponin content reduced on germination when compared with raw seeds. Saponins were found to be responsible for haemolysis of erythrocytes and causing various health-related issues (Valadez-Vega et al., 2022). Phytic acid another anti-nutritional factor present in amaranth forms insoluble stable conjugates by chelating trace elements in gastro-intestinal tract and ultimately reducing mineral element bioavailability by the human body. An effective way to overcome the reduced mineral element bioavailabilty is germination. Germination increase phytase and phosphatases activity which is responsible for hydrolysis of phytate complex and inositol promoting the seed growth. Hence, increase the absorption rate of mineral elements in the body (Liu et al., 2022).
5.2.5 Health Benefits of Amaranth Sprouts These improved chemical composition of amaranth sprouts makes it a good nutritional food for health promoting purposes and can prevent the human population from the curse of malnutrition also the world today need nutrient enriched grains. Therefore amaranth sprouts are wonderful source in providing health benefits. 5.2.5.1 Antioxidant Activity The antioxidant activity of any food product is generally determined by 1,1-diphenyl-2-picryl-hydrazyl antioxidant assay (DPPH), ferric reducing antioxidant power (FRAP), and 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid radical scavenging assay (ABTS) (Aloo et al., 2021). Different amaranth seed species possess various phenolic compounds which were determined by Folin–Ciocalteu method and found approximately 39.17 mg/100 g in A. caudatus and 56.22 mg/100 g
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in A. Paniculatus species (Klimczak et al., 2002). Amaranth grain consumption (155 and 310 g/kg of diet) help in restoring of enzymes and showed impact on oxidative stress by the decrease in malondialdehyde Although amaranth seed contains high concentration of phenolic compounds but sprouts being emblematic source of flavonoid rutin proves to a have a great level of antioxidant activities (Maurya & Arya, 2018). However, same species of amaranth sprouts from geographical region shows antioxidant activity higher in Amaranth cruentus v. Aztec lower in Amaranth cruentus v. rawa (Paśko et al., 2009). Amaranth oil also serves as a reservoir of antioxidant properties as it contains high content of unsaturated fatty acids like oleic acid, linoleic acid, and a little concentration of linolenic acid, and squalene. 5.2.5.2 Hypocholestrolemic Effect Various studies have been conducted to find out the effect of amaranth on the blood cholesterol level and it came out that amaranth helps in modulation of serum cholesterol levels, because of unsaturated fatty acids, soluble fibre, amino acid profile of amaranth protein and a number of phytochemical (phytosterols, tocotrienols, tocopherols, and squalene) present in it. For this purpose, a number of animal trials have been carried out (hamsters, rats, rabbit etc.) using different fraction of amaranth grain and found that amaranth grain as well as oil reduces the blood and liver cholesterol in humans. It is also reported that squalene increases the fecal excretion of steroids by interfering with the cholesterol absorption. Animal trial on rabbit fed with hypercholestrol diet and amaranth extract shows that LDL, TC, TGS, apolipoprotein, oxidised LDL and C-reactive protein decreased whereas apo-A and HDL cholesterol decreased as compared to diet comprising of only high cholesterol diet (Chmelík et al., 2019). On germination for 3 days concentration of linoleic, linolenic and oleic acid increased whereas concentration of palmitic acid decreases. It can be concluded that increase in oleic acid in amaranth sprout is responsible for a reduction in plasma cholesterol, cancer inhibition etc. which can be helpful in maintain cholesterol (Corzo-Ríos et al., 2021). 5.2.5.3 ACE Inhibitory Capacity In a research conducted to know the ace inhibitory activity in amaranth protein hydrolysate with alcalase. Result showed that protein hydrolysates with alcalase have better ACE inhibitory activity as compared to amaranth protein concentrate (Tiengo et al., 2009). Similar study conducted using amaranth sprout protein to explore antihypertensive activity using ACE inhibition resulted in IC50 0.24 mg/ml after digestion which was 0.67 before digestion (Aphalo et al., 2015). Therefore, amaranth sprouts have also been found to link with ACE inhibitory compounds in providing health stimulating food.
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5.2.5.4 Anti-tumour Effect Amaranth sprouts bear a great potential in improving the nutrition security for people around the world. One of the several reason for chronic disease like cancer is improper diet which with time of 10–30 years gets transformed into detectable tumors therefore the diet of a person fully depends for ailments. For this reason, studies investigated that amaranth lectins can bind to the tumor factors (TF-binding lectin). These TF-binding lectins play a role in diagnosis of malignant intestinal tumor activities (Maurya & Arya, 2018). Protein isolate of amaranth seed possess anti tumor properties as investigated by (Barrio & Añón, 2010). According to the research protein isolate of amaranth seed was used against four different cell lines (MC3T3E1, UMR106, Caco-2, and TC7), and the result showed different efficiencies against different cell line. Studies related with amaranth sprout anti-tumor properties are less explored but being a potential source of protein, these sprouts protein can be further investigated for their anti-proliferative effect. 5.2.5.5 Anti-inflammatory Action Sprouts are a potent source of many biologically active compounds which divulge their anticancer and anti-inflammatory activities through transduction pathway in cells. This was proven from an investigation that Amaranth sprouts during their growth were supplemented with selenium trace element 10 mg/ml 15 mg/ml in the form of sodium selenite. Amount of betacyanin in amaranth species was correlated with selenium doses, as with increase in selenium concentration betacyanin concentration marked a reduction. This study, concluded that betacyanin and selenium help in the prevention of NFkB translocation to cell nucleus, thereafter exhibiting anti- inflammatory effect by decreasing interleukin six production in cell culture of activated RAW 264.7 macrophage with ample health benefits listed above, amaranth also possesses numerous other health-promoting factors such as anti-obesity, anti- diabetic, gluten-free diet for celiac disease, and relief from skin related issues. To sum up, amaranth seeds, and sprouts acts as overall health maker (Tyszka-Czochara et al., 2016).
5.3 Quinoa Quinoa is one of the most popular pseudo cereal (Amaranthaceae family) which was first domesticated about 5000 years ago in Andean region of South America. It was cultivated by Incas emperor for its high nutritional, health beneficial properties, its ability to grow in salty soil and drought tolerance area. The Incas emperor considered it as “mother of all grain” due to its use for medicinal purposes. Quinoa was cultivated only in eight countries in 1980; later it spreaded to more than one hundred countries in 2021. 80% of total quinoa comes from south America including Bolivia,
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Ecuador, and Peru (Pathan & Siddiqui, 2022). The consumption of sprouted quinoa started in Korea, China, and Japan but later it gained more demand to other countries, including United States, Australia, and Europe. Over the last few decades, the production and consumption of quinoa also rapidly increased in western countries due to consideration of exotic and dietetics healthy foods (Singh & Singh, 2016). Quinoa seed contains a high quantity of protein, vitamin, minerals, phenolic compounds, and flavonoids and also contain sufficient quantity of essential therapeutic components such as squalene, saponins, fagopyritols, polyphenols, and phytosterols (Arneja et al., 2015). Sprouting of quinoa is a simple process which enhance the nutritional as well as functional value. In germination process, several enzymes are activated as well as are enabled to mobilize chemical compounds increasing mineral bioavailability, protein digestibility, antioxidant activity, and vitamin content. Regular consumption of sprouted quinoa contributes to health promotion. It reduces the risk of some chronic diseases such as certain cancers, inflammatory bowel disease, neurodegenerative disorders, arthritis, ischemic stroke, and cardiovascular due to having nutritional components such as amino acids, vitamins, minerals, and peptides.
5.3.1 Quinoa Seeds and Sprouts: An Overview In the last decade, the quinoa seed and sprouts have become more popular and widely consumed as a functional food diet for both ordinary people and those having celiac disease or allergies due to various health benefits such as the high amount of amino acids, vitamins, trace elements, fibre, flavonoids and phenolic acids. The nutritional composition of quinoa is shown in Table 5.3 (Ng & Wang, 2021).
Table 5.3 Nutritional composition of Quinoa seeds per 100 g
Chemical composition Fat Carbohydrates Protein Minerals K Mg Ca P Fe Mn Cu Zn N
Quinoa seeds (per 100 g) 6.0 62.1 13.1 (mg/100 g) 956.4 372.7 93.7 398.6 10.5 1.9 2.8 1.6 16.0
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5.3.1.1 Carbohydrate Quinoa starch (58.1–64.2%) mainly consist of amylose (∼10%) and amylopectin (90%) (Schoenlechner, 2017). The starch of quinoa had more solubility and digestibility due to a higher surface area for binding of water molecules and more enzyme digestion because of many branches (Selma-Gracia et al., 2020). Quinoa is rich in fibre (8–13%) compare to corn and rice (Tanwar et al., 2019). The quantity of insoluble fibre and soluble fibre ranged from 10% to 14%, and from 1.3% to 6.1% respectively. The variation of fibre composition is mainly influenced by growth conditions and genotypes. Homogalacturonans are the key ingredient of insoluble fibre and soluble fibre part mainly consists of arabinan and homogalacturonans (Zhu, 2020). 5.3.1.2 Protein Quinoa is a protein-rich food containing all kinds of essential amino acids, such as isoleucine and lysine. However, most cereals and grains do not have a sufficient quantity of isoleucine and lysine (Comino et al., 2013; James & Lilian, 2009). Normally, quinoa contains protein in the range of 12–23% and out of them, globulins (37%) and albumins (35%) are the main components (Dakhili et al., 2019). Bioactive peptides are present in quinoa as fragments of protein with some biological activities related to health benefits such as immunomodulatory, antioxidant, antihypertensive, and antimicrobial (Sánchez & Vázquez, 2017). Lunasin, a peptide in quinoa, has an important role in preventing cancer. Some other peptides such as QFLLAGR and ASPKPSSA are found in quinoa showing high free radical scavenging activity and iron chelating activity (Zheng et al., 2019). Quinoa possesses biological value (BV) of 73% which is nearly equal to biological value of beef i.e. 74% and also greater than corn (36%), wheat (49%), and rice (56%) due to the higher digestibility of quinoa protein (Gordillo-Bastidas et al., 2016). Additionally, quinoa is mostly preferable for patients with celiac disease due to being gluten-free (Comino et al., 2013). In-vivo experimental studies confirm that 91.6% of the raw quinoa proteins was absorbable (Ruales et al., 2002). The quantity of essential amino acids in quinoa is presented in Table 5.4. 5.3.1.3 Lipids Quinoa is also a valuable source of health-supportive fat (5.3–14.5%), out of which 70–89.4% is unsaturated fat which help to reduce diabetes and inflammation (Gordillo-Bastidas et al., 2016). About 28% unsaturated fatty acids of quinoa are made of oleic acid which is very supportive for our heart. About 5% unsaturated fatty acids of quinoa are made of alpha-linoleic acid which helps to reduce of the risk of inflammatory diseases. The quinoa oil mainly consists of ω-6 and ω-3 fatty
5 Amaranth & Quinoa Sprouts Table 5.4 Composition of Essential amino acids in quinoa protein (mg/g)
139 Essential amino acids Tryptophan Lysine Phenylalanine and tyrosine Valine Threonine Isoleucine Histidine Leucine Methionine and cysteine
Quinoa protein (mg/g) 8 51 74 45 30 37 25 64 21
acids which help to reduce the risk of breast cancer, cardiovascular disease (CSD), and gastrointestinal cancer (Gordillo-Bastidas et al., 2016; Graf et al., 2014). 5.3.1.4 Minerals Quinoa contains various kind of minerals such as zinc, calcium, copper, magnesium, and iron in higher quantity as compared to other cereal such as wheat, barley, oats, rye, triticale, and rice (Filho et al., 2017). The quantity of minerals in quinoa presented in Table 5.1. After germination, the quantity of iron, zinc, and calcium in quinoa was increased about 39.43%, 20.25, and 49.04% respectively (Darwish et al., 2020). According to James and Lilian (2009), Daily consumption of 100 g of quinoa can fulfil an adult’s daily requirement of iron, magnesium, copper, and manganese, and also meet 40–60% of adult’s daily needs of phosphorus and zinc. 5.3.1.5 Vitamin Quinoa is also a rich source of vitamin including vitamin C, thiamine, vitamin B5, vitamin B6, and folic acid (Gordillo-Bastidas et al., 2016; Schoenlechner, 2017) which helps to maintain the proper functional activity of brain and muscle cells. Folic acid is essential to reduce the risk of neural birth defects, especially for pregnant women. It also contains an adequate level of vitamin E, which acts as an antioxidant. Consumption of 100 g of quinoa daily can meet adult’s daily requirement of folate and vitamin B6, however, it can meet 40% of total need of riboflavin content of an adult (James & Lilian, 2009).
5.3.2 Germination of Quinoa Grains The germination process starts by consuming of water by the dry seed and finishes with the development of the embryonic axis. This technique was carried out to soften the kernel, enhancing the nutritional value as well as reducing the ant
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nutritional factors. Briefly, during the germination process: (i) During germination, dry seed rapidly absorb water and terminate the embryo axis elongation, (ii) this step is associated to activate a strong metabolic reaction by up taking limited amount of water (iii) this step involves with cell elongation for completing of germination. During the germination process, dry quinoa seed regain metabolic, hastily restore metabolic activity, remobilization, accumulation, and degradation, leading to several changes in the edible products such as biochemical, nutritional and sensorial changes. Various factors such as temperature, light, humidity, and seed varieties affect germination as well as change of nutritional value (Miyahira et al., 2021). Due to fault in handling operation, sprouts possess shorter shelf life. Therefore, it should be stored carefully.
5.3.3 Changes in Chemical Composition During Germination Quinoa requires a short time of about 4–5 h for germination compared to other grains, which require at least 12–14 h. During the germination process, the chemical composition (Carbohydrate, lipid, proteins, polyphenols, vitamins, minerals, antioxidant activity, and antinutrients) of quinoa seed changes due to reactivate the seed metabolism, resulting in degradation of macronutrients and anti-nutritional compounds. 5.3.3.1 Carbohydrates The germination process involves to change the carbohydrate content by increasing reducing sugars (RS), total soluble sugars (TSS), and decreasing the starch content. In early stage of germination, α-amylase is not synthesized, but with increasing the germination time, the synthesis rate of amylase rapidly increases, which leads to increased conversion of starch to glucose, sucrose, and dextrin. These glucose, sucrose, dextrin on further oxidation tend to yield energy for growth and development of the embryo (Elkhalifa & Bernhardt, 2010). 5.3.3.2 Proteins Germination has also an important role in increasing the protein concentration of quinoa. Total quinoa protein content increased from 9.5 to 25.9.0 g/100 g dry weight after 72 h germination. The reason for increasing protein content due to the mobilization of reserve nutrients and generation in grain as well as loss of dry matter through respiration (Pilco-quesada et al., 2020). The proteins are hydrolysed during germination, and consequently, their digestibility is also improved. Proteins having a high molecular weight (>24 KDa) were degraded enzymatically and decreased
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their molecular size, which lead to an increased protein digestibility of about 15–42% after germination (Jimenez et al., 2019). 5.3.3.3 Lipids The lipid content of quinoa was significantly decreased during germination. After 72 h of germination, the total lipid content was found to be decreased from 15.1 to 7.5 g/100 g (dry weight) in quinoa. During the germination stage, the lipid content in quinoa converts into sucrose and 25% of the lipids of quinoa are hydrolysed to meet the energy requirement for respiratory activity (Pilco-quesada et al., 2020). At the end of germination, total lipid and bound lipid were increased by 5% and 100%, respectively. However, free lipid was decreased by 33% (Omary et al., 2012). 5.3.3.4 Vitamins and Mineral Germination may also improve nutritional value by increasing the availability of minerals in quinoa due to a decrease of anti-nutrients, including phytic acid. Phytic acid binds the mineral, protects it from extraction, and makes it insoluble complexes. Germination increased the quantity of chromium, zinc, copper, and strontium by 15, 17, 24, and 30% and decreases in manganese, molybdenum, and lithium, and of 7, 12, and 50%, respectively using tap water. No significant variation was found for vanadium, cobalt, arsenic, iron, and selenium (Omary et al., 2012). 5.3.3.5 Bioactive Compounds About twenty-one phenolic compounds such as hydroxycinnamic acids, flavonols, hydroxybenzoic acids are present in quinoa (Pilco-quesada et al., 2020). Germination also increased the polyphenol contents and antioxidant capacity of quinoa. Alvarez- Jubete et al. (2010) mentioned that the total phenolic components of quinoa increased from 110.0 mg/100 g to 153.00 mg/100 g and antioxidant from 26.05 to 38.75% after 48 h germinations. Vitamin ‘C’ also was found to be increased from 10.56 to 13.11 mg/100 g after germination. 5.3.3.6 Anti-nutritional Compounds Germination of quinoa significantly decreases anti-nutritional factors such as saponin, phytates, and inhibitory proteases (Jimenez et al., 2019). After germination, phytic acid and saponin were decreased from 63.67 mg/100 g to 44.87 mg/100 g and from 0.65 to 0.45 g/100 g respectively. As phytic acid comes from phosphorus, therefore, it might have broken down for phosphorous utilisation during germination which lead to decrease the phytic acid. Similarly, the washing and soaking
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phase of germination decreased saponins and tannin through the leaching process by water (Padmashree et al., 2019).
5.3.4 Health Benefits of Quinoa Sprouts 5.3.4.1 Antioxidant Properties The quinoa seed contains many phenolic compounds (approx. 23), which act as antioxidants. Ferulic acid and quercetin are the most widely found phenolic compounds in quinoa which are associated with preventive action against degenerative diseases linked with the free radical change. Quinoa also enhances the antioxidant capacity of the heart, blood (plasma), kidney, lung, testis and pancreas and acts as moderate protective agent against lipid peroxidation (Arneja et al., 2015). Numerous evidences have demonstrated that increased oxidative stress pursue many adverse effects on human health such as CSD, Alzheimer disease, cancer and depression (Al-Qabba et al., 2020; Zheng et al., 2019). Some antioxidants, like polyphenol, flavonols present in quinoa that helps to lessen the effect of oxidative stress and reduces the threat of cardiovascular and Alzheimer disease (Holland et al., 2020). Studies showed that flavonoid and phenolic compounds of quinoa seed contributes to the higher yield of TPC (total phenolic contents) on dry weight basis i.e. 102.85 mg GAE/100 g, which encompasses 28.8% of radical scavenging capacity (DPPH) (Ahmed et al., 2020). Apart from phenolics, bioactive peptides also have the potential to neutralize free radicals by electron donation and inhibit lipid oxidation (Ayyash et al. (2018). Polysaccharides, Polyunsaturated fatty acids (PUFA), and saponins are found in quinoa to favour antioxidant capacity of quinoa. On the other hand, antioxidant activity of quinoa is also found to be altered by germination, fermentation, cooking, and milling process. Moreover, it is also shown that quinoa possess better antioxidant activity as compared with grains (Al-Qabba et al., 2020). Therefore, quinoa could serve as a functional food due to strong antioxidant activity for prevention of diseases. 5.3.4.2 Effect on Cholesterol From the past studies, it is revealed that higher triglyceride and low-density lipoprotein cholesterol (LDL) are responsible for cardiovascular disease and higher level of high density lipoproteins proved to be beneficial for human health as it removes cholesterol from the body. Various In-vivo studies come up with the fact that, quinoa seed positively affect the human cholesterol level as it significantly reduce low- density lipoprotein cholesterol (LDL-C) and triglycerides (TG) level and augment the high-density lipoprotein cholesterol (HDL-C) levels (Farinazzi-Machado et al., 2012; Navarro-Perez et al., 2017). Furthermore, quinoa protein also help to bind bile acid which helps to reduce absorption of lipids from food and promotes the
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reabsorption of serum lipid, ultimately lowering the serum cholesterol. Quinoa, also inhibit the activity of cholesterol-synthesizing enzyme, 3-hydroxy-3-methylglutarylcoenzyme A (HMG-CoA) reductase. 5.3.4.3 Effect of Consuming Quinoa on Blood Sugar Quinoa is a potential source of phenolic, 20-hydroxyecdysone (20HE) tocopherols, proteins, bioactive peptide and fiber which play vital role in preventing diabetes (Ayyash et al., 2018). Peptides of quinoa have important role to inhibit α-amylase, blood sugar, dipeptidyl-peptidase-4 (DPP-IV), and α-glucosidase (Guo et al., 2020). This control of blood glucose level by α-amylase and α-glucosidase works by decreasing the carbohydrate digestion rate. Also, high amount of protein in quinoa gradually slow down digestion and gastric emptying process (Cisneros-Yupanqui et al., 2020). 5.3.4.4 Effect of Quinoa on Liver Health The antioxidant components of quinoa are related with the liver health as quinoa possess anti-inflammatory and antioxidant properties defined as its hepatoprotective potential. Liver is largely found responsible, for maintaining metabolic reactions when ROS (reactive oxygen species) and free radicals are formed. Phenol compounds, polysaccharides and vitamins (fat-soluble) reduce stress and ultimately prevent the liver from any oxidative injury or damage (Ahmed et al., 2020; Ali, 2019). In-vivo trials conducted on high-cholesterol-diet-induced rats, infected fish, cyclophosphamide-poisoned rats, high-fructose-diet-induced rats and healthy rats indicates that quinoa rich diet improved the liver functioning in all the experimental species (Fotschki et al., 2020; Wahba et al., 2019). Ahmed et al. (2020)) mainly demonstrated that the aspartate aminotransferase (AST) and Serum alanine aminotransferase (ALT) are the two essential parameters to examine liver function. Quinoa diet significantly reduces serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) and levels as higher levels of AST and ALT causes liver damage. Therefore, regular consuming quinoa can reduce ALT and AST levels which could be beneficial to maintain the liver function. 5.3.4.5 Gluten Free To produce and consume gluten free foods is now a necessity specially for those who have not ability to digest gluten protein (wheat, barley, rye etc.) and their bodies creates certain inflammatory disorder in small intestine affecting the human health generally termed as Celiac disease. Therefore, quinoa seeds can serve as an alternative to gluten protein foods for celiac patients or gluten intolerance people (Arneja et al., 2015).
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5.4 Application of Amaranth and Quinoa Sprouts in Food Industries Knowledge of pseudocereals for human health and nutrition in the past few years has alarmed its use for the production of novel and innovative food for maintaining health. In this respect, food industries find their way to develop nutritionally rich foods incorporated with pseudocereals like amaranth, quinoa, buckwheat etc. Use of pseudocereals can be done as starchy grains, milled and fractionated seed crop. Depending on composition and botanical classes these gluten-free crops may require different processing conditions. Also, their small size, gluten free nature, small starch granule makes this task more challenging and ultimately resulting in differences in technological properties. Various considerations must be focussed before incorporating pseudo cereals into food products; Pseudocereals possess different botanical structure and seed morphology – Due to their small size, these seed crop requires different milling conditions. To acquire different flour fractions wide range of milling procedure are needed and therefore milled as wholemeal flour, which is an advantage because of its high nutritional value. Recently food industries are using such floors for the production of gluten-free products such as bakery products, and pasta due to enhancement in nutritional and bioactive components (proteins, flavonoids, peptides, vitamins, fatty acid, phenolic acid, amino acid, unsaturated fatty acid and dietary fibres) during germination. It is assumed that gluten-free production of food will be increased at an annual growth rate 9.1% from 2019 to 2025 (Graziano et al., 2022). In case of amaranth seed, hull is denser and compact which is another factor to be considered. Likewise, popped maize kernels, amaranth seeds can also be popped when provided with dry heat. These popped amaranths can further be used for direct consumption or milled into flour; Gluten free properties – pseudocereals being free from gluten makes difficult to produce leavened product and hence requires some processing steps in addition necessary steps that follows; Starch properties – Amylose content, starch composition, structure of starch kernel results in alteration of physicochemical properties (Valadez-Vega et al., 2022). Used amaranth sprouts as ingredients for the food formulations and observed that sprouts served high concentrations of soluble protein, total flavonoid content, total phenolic content, total anthocyanin and antioxidant activity. Gluten-free eggless cake is prepared using sprouted amaranth flour for patients suffering from Celiac disease and as an alternative to refined flour with high nutritional composition (Sandoval-Sicairos et al., 2020). Germinated amaranth found its application in the preparation of dough for bread making in combination with wheat flour with little change in hardness and rheological properties (Agrahar-Murugkar et al., 2018).
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5.4.1 Gluten-Free Cereal Based Foods Market scenario of gluten free products is currently under process as the products available are of low nutritional quality. As quinoa does not possess gluten, it could be helpful for the people who are suffering from wheat allergy or celiac disease. Therefore quinoa can be find its wide application in producing gluten free crackers, biscuits, bread through its germination. Studies revealed that replacing potato starch for bread with quinoa or amaranth flour results in higher nutritional value (fiber, protein, magnesium, calcium, iron, vitamin E, and zinc (Valcárcel-yamani & Caetano, 2012).
5.4.2 Composite Flours in Development of Bakery Items and Pasta Technology of composite flour indicates to the mixing process of wheat and non- wheat flour (quinoa flour, amaranth) containing high amount of protein for production of various bakery products and pasta. Recently in food industry, quinoa is used to make pasta, bread, and other bakery products for improving quality (Valcárcel- yamani & Caetano, 2012) but reduces volume, increases hardness, and confers a dark colour and bitter taste (Haros & Sanz-Penella, 2017). These negative characteristics depend on the percentage of quinoa flour used that was not perceived up to 25% replacement of wheat flour (Iglesias-Puig et al., 2015). While the use of germinated quinoa flour (15–25%) increase the content of polyphenols and antioxidant properties in dough and also reduce the hardness and bitterness (Ballester-S’anchez et al., 2019). The mixing of wheat and quinoa improved the bread quality such as no aerated crumb structure along with good sensory properties. The replacement of quinoa flour (7.5–10%) with wheat flour significantly increased the loaf volume of bread, but more than 15% substitution distinctly decreased it. Addition of quinoa to the extruded products yield a product with high fiber, ash, protein and few amino acid in comparison to 100% pure wheat based products. Products comprising of quinoa showed good consumer acceptance. Likewise, germinated quinoa flour enhanced the nutritional value of pasta (higher protein and amino acid contents, and lower amounts of phytate) and reduce the pasta firmness (Demir & Bilgiçli, 2020).
5.5 Future Perspective Due to upcoming consumer demand for sprouted pseudocereal food products (Amaranth, quinoa, buckwheat) with enhanced health benefits and nutritional value, it becomes necessary to fulfil the demands of the customer, which brings certain challenges for the food industry like germination process optimization for
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production (stage of growth and conditions of germination), composition, safety, and nutritional value for the newly developed product. Moreover, research is required to analyze pre-harvesting and post-harvesting technology to avoid microbiological risks. Further, the processing effect on the bioactive compounds will give the conditions required to attain the desirable and most adequate product. Future studies on the exploitation of pseudocereals emphasize developing functional food by-products for health and wellness (Benincasa et al., 2019; Giménez-Bastida et al., 2015). Therefore, utilization and value addition to these sprouted pseudo-cereals in bakery industries have been recognised as the major area of research. However, consistent effort and focused research are still required to bridge the gap between the production and commercialization of pseudo-cereal-based convenient smart foods (Patil & Jena, 2020).
5.6 Conclusion According to vast literature survey it can be concluded that on germination of pseudocereals their nutritional value enhanced tremendously leaving behind anti- nutritional factors (trypsin inhibitor, phytate, tannins, etc.) which suppress the effect of various nutrients. Starch reserves gets mobilised by the action alpha amylases forming pinholes, brings changes in amino acid composition, saturated and unsaturated fatty acid content increases along with rise in bioactive compound. Therefore, almost all nutrients are present in adequate amount and higher antioxidant activity making it a functional food (Benincasa et al., 2019; (Paucar-Menacho et al., 2022). It is researched that European and American food industries are not permitted to disclose some major benefits of sprouted pseudocereals, like bioavailability of micronutrients. Also, processing may impart a unique flavour or desired differentiated flavour (Pagand et al., 2017). All these factors allow the food manufacturers to widely produce sprouted pseudocereals based products.
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Thapa, R., & Blair, M. W. (2018). Morphological assessment of cultivated and wild amaranth species diversity. Agronomy, 8(11), 272. https://doi.org/10.3390/agronomy8110272 Tiengo, A., Faria, M., & Netto, F. M. (2009). Characterization and ACE-inhibitory activity of amaranth proteins. Journal of Food Science, 74(5), H121–H126. https://doi. org/10.1111/j.1750-3841.2009.01145.x Tyszka-Czochara, M., Pasko, P., Zagrodzki, P., Gajdzik, E., Wietecha-Posluszny, R., & Gorinstein, S. (2016). Selenium supplementation of amaranth sprouts influences betacyanin content and improves anti-inflammatory properties via NFκB in murine RAW 264.7 macrophages. Biological Trace Element Research, 169(2), 320–330. https://doi.org/10.1007/s12011-015-0429-x United States Department of Agriculture (2011). National Nutrient Database for Standard Reference. Food Composition Database. Retrieved May 16, 2022, from http://ndb.nal.usda.gov/ Valadez-Vega, C., Lugo-Magaña, O., Figueroa-Hernández, C., Bautista, M., Betanzos-Cabrera, G., Bernardino-Nicanor, A., et al. (2022). Effects of germination and popping on the anti- nutritional compounds and the digestibility of amaranthus hypochondriacus seeds. Food, 11(14), 2075. https://doi.org/10.3390/foods11142075 Valcárcel-yamani, B., & Caetano, S. (2012). Applications of Quinoa (Chenopodium Quinoa Willd.) and Amaranth (Amaranthus Spp.) and Their Influence in the Nutritional Value of Cereal Based Foods. Food Public Health, 2(6), 265–275. https://doi.org/10.5923/j.fph.20120206.12 Wahba, H. M. A., Mahmoud, M. H., & El-Mehiry, H. F. (2019). Effect of quinoa seeds against cisplatin toxicity in female rats. Journal of Advanced Pharmacy Education & Research, 9(3), 47. Zheng, Y. J., Wang, X., Zhuang, Y. L., Li, Y., Tian, H. L., Shi, P. Q., & Li, G. F. (2019). Isolation of novel ACE-inhibitory and antioxidant peptides from quinoa bran albumin assisted with an in silico approach: Characterization, in vivo antihypertension, and molecular docking. Molecules (Basel, Switzerland), 24(24), 4562. https://doi.org/10.3390/molecules24244562 Zhu, F. (2020). Dietary fiber polysaccharides of amaranth, buckwheat and quinoa grains: A review of chemical structure, biological functions and food uses. Carbohydrate Polymers, 248, 116819. https://doi.org/10.1016/j.carbpol.2020.116819
Chapter 6
Oat and Kamut Sprouts Pooja Kesarkar, Papiha Gawande, and Yogesh Gat
6.1 Oat Sprouts In terms of cereal production worldwide, oat comes in approximately sixth position after wheat, rice, maize, barley and sorghum. Since many decades, oat grain has considered as staple food and feed. An annual, oats can be planted in the spring or the fall and harvested in the late summer (for early autumn harvest). In the Himalayan foothills, oats are grown, including in the Indian state of Himachal Pradesh, where they are known locally as “jau”. Oat is a good source of protein, minerals and fibre. For individuals in marginal ecologies all across the developing globe, as well as in industrialised economies for specialised needs, oats continue to be a significant grain crop. Oats are cultivated for use as a grain, bedding straw, hay, forage and fodder, chaff, haylage and silage in various parts of the world. Oat crops are still primarily used for grain feed for livestock, which accounts for an average of about 74% of global utilisation. Compared to other small grain cereal crops, oats are more adaptable to a variety of soil types and can thrive on acidic soils. Oat is mostly produced in cool, wet regions, and from the time the head first appears until it reaches maturity, they can be sensitive to hot, dry conditions. Due to these factors, the majority of the world’s oat production occurs between latitudes 20–46°S and latitudes 35–65°N, which includes Finland and Norway. Although autumn sowing is used along greater altitude locations, such as the Himalayan Hindu Kush range, and in areas where summers are dry and hot, spring sowing accounts for the majority of global production. Short-season to mid-maturing oat cultivars are typically cultivated in locations with harsh winters, such as Scandinavia, the northern US states, Canada, and higher altitude tropical climates. Oats are often sown in the spring, P. Kesarkar · P. Gawande · Y. Gat (*) Department of Food Engineering and Technology, Institute of Chemical Technology, Mumbai, Maharashtra, India e-mail: [email protected] © The Author(s), under exclusive license to Springer Nature Switzerland AG 2023 I. Majid et al. (eds.), Advances in Plant Sprouts, https://doi.org/10.1007/978-3-031-40916-5_6
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winter, or autumn in temperate countries, depending on the local climate, the need for crop rotation, the intended use, and other agricultural methods. Oat flour, oatmeal, oat flakes, and oat bran are all edible forms of oats that are used as morning cereals and as ingredients in other foods. One of the healthiest grain cereals, oats are rich in protein and fibre. Oats that have been rolled into flakes typically have more protein than other cereal grains. As compared to other grains, oats are more adaptable in India, especially in the western and north-western parts of the country due to their superior growing environments, quick regrowth, and higher nutritional value.
6.1.1 Composition of Oat Oat, Groats, Haber, Hafer, Avena, Straw, and Oatmeal are all names for the cereal grain Avena sativa L. (Gramineae), which is cultivated for its seed. The oats are grown all over the world. The Sanskrit word “avi,” which means “sheep,” or “avasa,” which means “foodstuff,” are the roots of the word “Avena”. Oats are often regarded as a secondary crop because they were first used to make bread in English bakeries. Oats were originally a weed of wheat and barley that eventually led to their domestication. They continue to be highly valued in Scotland as a staple of the country’s cuisine, and they were formerly a common food source for many people before coffee and bread took their place. Table 6.1 comprises nutritional components in oats. Oats are divided into several categories according to their level of processing. For human consumption, the hard outer hull of whole oats must be removed. The term “groats” refers to hulled oats. The chemical furfural is derived from oat hulls. Oat groats preserve the outer bran layer of the kernel while removing the tough, unappealing outer hull from the entire oat grain. The process of making steel-cut oats commonly referred to as pinhead oats and occasionally as coarse or rough oatmeal, involves running groats through steel cutters that separate each one into 3–4 pieces. Groats are steamed before being flattened with a roller to create rolled oats. Except for being steamed for a longer time and being rolled more thinly, instant oats are manufactured like rolled quick-cooking oats. The oats are milled to produce oat flour, which is typically offered in three grades: coarse, medium, and fine. Table 6.1 Nutritional components in Oats
Nutritional component Protein Beta-glucan Thiamine Riboflavin γ-aminobutyric acid Free phenolics Antioxidant capacity
Units 10.7 g/100 g 2.1 g/100 g 687.1 μg/100 g 218.4 μg/100 g 54.9 mg/100 g 507.4 mg GA/100 g 1744.3 mg TE/100 g
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6.1.1.1 Phytoconstituents Carbohydrates The sugar (glucose, fructose) 3–4%, mucilage (β-glucan), pentosan, kestose, saccharose, pentosans, neokestose, neobifurcose, bifurcose acid were present in oats. As a mixed-linkage polysaccharide containing D-glucose units, oat β-glucan is a significant source of dietary fibre for oats. The D-glucopyranosyl units in β-glucan are linked together by 1–3 or 1–4 linkages. The molecule’s normal structure is altered by the 1–3-linkages, making it fluid and pliable. The majority of glucan is composed of cellotetraosyl and cellotriosyl units, although it also contains cellulose like − 1-4 linked glucose units. The oat grain has the highest concentration of starch. It has typical gelatinization capabilities and contains between 25% and 30% amylase. In comparison to other cereals, oat starches are clearer, more elastic, less rigid, stickier, less likely to retrograde and stickier. Proteins Oat is very high in protein content and is only source that contains avenalin, as the primary (80%) storage protein. The disulphide-linked and isomers of the unreduced heterogenous proteins have molecular weights between 53,000 and 58,000. Avenin, a prolamine, is the minor protein found in oats. Oat protein is almost as high-quality as soy protein, which the WHO claims is comparable to the protein found in meat, milk, and eggs. The oat kernel (groat) without the hull has the greatest protein level of any cereal, ranging from 12% to 24%. Lipid Lipids were isolated from a variety of oats, including triglycerides, steryl esters, glycolipids, free fatty acids and phospholipids. All cultivars contained almost the same amount of phospholipids. Compared to glycolipids or phospholipids, triglycerides had more oleic acid and less palmitic acid. According to reports there are 11 phospholipids and 9 glycolipids present in specific oat varieties (Holmback et al., 2001). Fibres Oats include both soluble and insoluble fibres. According to reports, eating a diet high in fibre helps avoid several ailments. It contains a lot of soluble fibre in the form of glucan as well as cellulose and arabinoxylans. Oats have the highest concentration of soluble fibre. Oat promote slower digestion and a fuller sensation. A group of indigestible polysaccharides known as D-glucans can be found in cereals,
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algae, mushrooms, yeast, barley and bacteria. However, oat-glucan is a soluble fibre. Despite the fact that the soluble glucan’s fibres are not digested in the small intestine, it increases the viscosity of the food bolus, delaying gastric emptying, improving gut fill, and delaying nutrient absorption. This leads to the formation of larger, more rapid stools. The physiological effects involve the synthesis of gut hormones and insulin. Glucans have a proven ability to lower cholesterol. Oatmeal provide 3 g of soluble fibre per day. Whole grains have long been used in nutrition as carriers for molecules like vitamins, microelements, and fibre in addition to other molecules. Cereal grains and glucans provide information on the glucan content present in various oat products. Wheat only has about 1% glucan, compared to oats, which have 3–7%. The potential use of glucan as a food component in functional dietary fibre is becoming more important. There are enough glucans in rolled oats, oat bran, and other oat products to be healthy. Numerous studies have shown how useful oats are for maintaining healthy arteries, blood pressure, and insulin levels. Vitamins Oats contain vitamins that are soluble in both water and fat. Oat seeds include the following vitamins: Vitamin A as 0.862 mg/100 g (mucilage); Beta-carotene: E/ tocopherol minimum 4:3; (tocopherol) 0.5–1.0. Minerals Oats contain a lot of minerals. Table 6.2 lists the mineral composition of green oats and oat seeds. Selenium, a vital cofactor of the significant antioxidant glutathione peroxidase, is also found in abundance in oats. Alkaloids Gramin, an indole alkaloid discovered from A. Sativa, is assumed to be the cause of the fruit’s mild sedative effects. Avenanthramides are phenolic compounds made of anthranilic and hydroanthranilic acids that are joined by an amide bond to one of Table 6.2 Mineral content of Oat
Mineral Calcium Magnesium Potassium Sodium Phosphorus Iron Zinc Manganese Copper
Mg/kg 682.85 1913.42 5590.74 143.73 4465.73 40.68 31.67 30.13 3.68
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many hydroxycinnamic acids. In oat, p-coumaric and hydroxyanthranilic acid, caffeic acids combine to generate the 3 most common avenanthramides. Gramine, an indole alkaloid discovered from A. sativa, is assumed to be the cause of the fruit’s mild sedative effects. Tocols Oat grain contains lipid-soluble tocols (tocopherols and tocotrienols), which are natural antioxidants also referred to as vitamin E. Tocotrienol, which is followed by tocopherol, is the primary tocotrienol present in oats. Only trace levels of other tocotrienols are present. The primary tocols present in oat are tocotrienols, which is more potent free radical scavengers than tocopherols. Saponins Among the monocots, triterpenoid saponins are rare. Avenacosides, avenacin from the root, and triterpenoid saponins. Avenacin biosynthetic genes may hold the key to creating cultivated kinds that are resistant to illness. A. sativa leaves have also been reported to contain avenacoside. Sterols Oat leaves contains sterols, sterol glycosides (SG), acylated sterol glycosides (ASG), and steroidal saponins. The majority of the sterol components were sitosterol, stigmasterol, cholesterol, cholesterol, 5 and 7 avenasterol, campesterol, campesterol, lophenol, stigmasterol, and 7-stigmasterol. There has been sufficient information about quantitative variation in several sterol groups present in oats.
6.1.2 Sprouting Process By metabolising carbohydrates, the embryo of oat generates a new plant during sprouting, resulting in the development of the radicles and coleoptile. As a result, the aleurone layer secretes hydrolases into the endosperm of the grain, including amylases and proteinases. As a result, the endosperm converts proteins and starch into glucose and other transportable sugars, peptides, and amino acids. In order to synthesise the first leaf and radicles, these substances are transported throughout the embryo to the growing regions. Sprouting only takes place in an aerobic atmosphere, at a temperature that is favourable, and with a moisture content that is high enough (>30%). Managing these parameters can have a direct impact on the biological activities in the grain.
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The process of germination involves the seed absorbing moisture and going through different metabolic processes to grow new roots and shoots. For the steeping and sprouting process, oat grains were rinsed under running water for 30 min before to the steeping and sprouting process in order to remove any microorganisms and prevent microbial growth. After that, the oat grains were steeped for 4.5 h wet steeping, 19 h air resting, and 4 h steeping at 20 °C in closed containers filled with water (covering all of the grains). The grains were drained and placed on a metal sheet after the steeping process. The steeped oat grains were wrapped in plastic wrap and kept in a climate cabinet at room temperature for several days. The grains were sieve-washed once day while they were growing. The water content was maintained constant during the washing procedure and check by moisture analyser. Many enzymes, nutrients, and genetic information in the seed’s endosperm and germinal disc are activated during germination, securing the maximum amount of nutrition. More specifically, as the germinal disc germinates, proteins change in quality. Enzymes are triggered during germination, and the softening of the grain may result in a better texture. Additionally, when carbohydrates become more digestible, the body can absorb nutrients more readily.
6.1.3 Nutritional Properties of Sprouted Oats Although oats don’t naturally contain gluten, their safety for people with celiac disease is still debatable because they can be easily contaminated by gluten- containing cereals in the field, or processing, during transport. Table 6.3 shows nutritional composition of oat grain VS sprouted grain. Avenins from different oat cultivars react differently to anti-gliadin antibodies, and some oat cultivars can trigger transglutaminase-2(TG2)-mediated events in vitro models of celiac disease due to genetic heterogeneity in oat cultivars. The usage of high-purity gluten-free oats derived from non-toxic varieties will enable the creation of secure gluten-free foods that support the health and nutrition of gluten-intolerant people. Sprouting is a useful technique for nutritional purposes due to the rise in protein, phenolic compounds, essential amino acids, and GABA during oat germination (Rozan et al., 2000). Additionally, sprouts satisfy the desires of contemporary consumers for all-natural, wholesome plant-based foods. Protein percentage was found to have slightly increased (by 11%), which is consistent with the dry weight losses through respiration (Aparicio-García et al., 2021). The considerable hydrolysis of oat proteins during germination is indicated by the weakening or disappearance of number of protein bands in sprouted oat powder in the electrophoresis gel. Protein, important amino acids (Met, Phe, Cys), riboflavin, minerals (Ca, Fe, Zn and Mg) and unsaturated fatty acids are all abundant in sprouted oat powder. Increased protease, amylase, and lowered lipase activities were seen in the sprouted oat powder, which may be used to improve its nutritional, sensory, and health- promoting properties.
6 Oat and Kamut Sprouts Table 6.3 Nutritional composition of germinated and un-germinated Oat grain
159 Nutrient/other constituents Water (g/100 g) Dry matter (g/100 g) Protein (g/100 g) Fat (g/100 g) Carbohydrates (g/100 g) Energy (Kcal/100 g) Fibers (g/100 g) Ash (g/100 g) Calcium (mg/100 g) Phosphorus (mg/100 g) Vitamin A (IU) Vitamin E (mg/100 g) Niacin (mg/100 g) Riboflavin (mg/100 g) Thiamine (mg/100 g) Vitamin C (mg/100 g)
Oat grain 7 50 7.5 2.1 32.9 181 5.9 1.6 32 180 0 0.9 0.9 0.1 0.2 0.2
Sprouted oats 404 50 10.5 2.6 21.4 151 13.1 2.0 119 254 3039 2.4 5.2 1.1 0.6 10.9
Source: Lasztity (1998)
The most common essential amino acid was lysine, followed by Val and Leu. Oat is rich in Lys, unlike most cereals, which have Lys as a limiting amino acid. The non-essential amino acid concentration was identical between the two, with the exception of Gly and Ala. The essential amino acid profile of sprouted oat demonstrated a discernible improvement in the levels of Met, Cys, and Phe while the other essential amino acids remained unaffected.
6.1.4 Use in Human Nutrition Because of oats’ dietary advantages and nutritional value, their use for human consumption has gradually expanded. The total dietary fibre and glucan content of oats play a major role in the health benefits. Oat protein is almost as high-quality as soy protein, which the WHO (World Health Organization) has determined to be comparable to the proteins in milk, eggs and meat. The oat kernel (groat) without the hull has the greatest protein level of any cereal, ranging from 12% to 24%. Several vitamins and minerals that are crucial for human health can be found in abundance in oats, along with soluble fibre, well-balanced proteins, and other nutrients. In comparison to other cereal grains, oats have a disproportionately high concentration of lipids and a significant amount of the important fatty acid linoleic acid. In addition, oats are a unique source of avenalumic acids (ethylenic homologues of cinnamic acids) and avenanthramides (N-cinnamoyl anthranilate alkaloids) which are not found in other cereal grains. Due to their antioxidant qualities and/or membrane-modulating properties (alk(en)resorcinol), all of these phenolic
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compounds have the potential to promote good health. Additionally, the soluble dietary fibre fractions of oats contain glucans, an antioxidant compound that also participates in gluco-regulation and lowers serum cholesterol levels in people (Krapf et al., 2019). Oats are consequently a crucial part of the diet for people with hypercholesterolemia. The majority of earlier studies in the literature showed that oats have high antioxidant properties. Oat grains contain soluble fibre called glucan, which has several useful functions and bioactive qualities. Its favourable effects on hypertension, dyslipidemia, obesity and insulin resistance are well-documented. The basis of glucan’s health advantages may lie in their capacity to ferment and their capacity to create extremely viscous solutions in the human gut. To increase the fibre content of food items and improve their health attributes, glucan’s suitability as a food additive is, therefore, being heavily considered. Growing quality oats has economic benefits for the farming community as well, thus this crop should be promoted. A key element of cellular walls is a carbohydrate called glucan. Because they contain significant levels of glucan, some microorganisms, including yeast and mushrooms, as well as cereals like oats and barley, are of commercial interest. These drugs boost immune function by modifying humoral and cellular immunity, which helps the body fight infections (bacterial, viral, fungal and parasitic). Additionally, glucan has anticoagulant and hypocholesterolemic effects.
6.1.5 Parameters Affecting Nutritional Quality During Sprouting 6.1.5.1 Seed Viability It is possible to think of the development and germination of seed sprouts as a set of successive occurrences that allow relatively low-moisture seeds to demonstrate an increase in metabolic activity that causes the protrusion production from the embryo. Due to their moderate resistance to harsh environmental circumstances, cereal seeds can maintain their viability for a considerable amount of time. During the malting process, oat rootlets were always longer than shoots, although their dry weight percentage was lower than that of shoots. Even though the respiratory losses grew steadily from 72 to 96 h, the pace of increase was less than it had been the previous 24 h. Dry matter losses from respiration significantly decreased after 96 h of germination. Malting oats had a very active life during the first half of germination, but the metabolism of the store materials predominantly produced energy release rather than the development of new organs. On the other hand, dry matter losses through the removal of rootlets and shoots became increasingly significant in the second half of germination.
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6.1.5.2 Availability of Water The first step of germination is the ability of seed to absorb water, and this ability is mostly governed by the seed coat permeability. The nature of seed, accessibility of the water in the surrounding environment, solutes concentration in solution, and other factors will all affect this imbibition. How well the seed can hold water and how much water is available to hydrate the seed during germination and later sprouting can be determined by the strength of this imbibition pressure. The protein consumes water predominantly during germination. As with cereal grains, seeds contain quite high concentrations of starch, but this won’t have a big impact on how much they expand. The swelling will also be caused by cellulose. Controlling the water supply is crucial to the success of cereal grain sprouting. The most important requirements for sprout production are proper moisture and temperature levels. 6.1.5.3 Gas Atmosphere The ambient air’s makeup has an impact on seed germination. The majority of seeds will begin to grow in the air that contains 20% O2 and 0.3% CO2, which is the typical concentration of gases in the atmosphere. But when the oxygen tension is raised above the usual 20%, some cereal grains will exhibit accelerated germination. If the tension of carbon dioxide is significantly raised, the majority of seeds do not germinate. Grain storage benefits from a very high carbon dioxide concentration because it suppresses germination. Nevertheless, rice appears to have an adaption for germination in an anaerobic environment, this is no need for rice to germinate. When seeds are sprouting in a container, the atmosphere around them can shift, which can result in lower oxygen levels and higher carbon dioxide levels. To ensure the best possible atmospheric conditions when growing sprouts, enhanced and frequent aeration generally improves germination and sprout development. 6.1.5.4 Temperature and Time The temperature and length of the sprouting process both affect the glucan content. After 3 days of sprouting, the glucan content decreases at all sprouting temperatures. The deterioration is most noticeable at 20 °C, which is the sprouting temperature. The glucan content falls somewhat between 10 and 14 °C for sprouting; however, this decrease is less pronounced at lower temperatures. Changes in ascorbic acid levels follow the progress of the sprouting process. In developing grain, ascorbic acid is required as a protective antioxidant. Oats’ increased vitamin C content during sprouting is the result of some mechanisms. Reactive oxygen species must be released to end the dormancy and begin the sprouting phase. These species do, however, cause oxidative stress in the cells as a result
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of their presence. Additionally, exposure to light stresses the grain, triggering the production of antioxidants. 6.1.5.5 Degree of Sprouting The qualities of the oat kernel are also impacted by the degree of sprouting. Radicles and coleoptiles are particularly affected by the increased levels of reducing sugars and ascorbic acid. In comparison to the grain without the coleoptile and radicles, the ascorbic acid concentration of radicles and coleoptile is 4 times higher. Therefore, it is crucial to remove the radicles and coleoptile from the grains in order to produce oat flour with a high nutritional content. The sweetness can be measured using the link between the degree of sprouting and the quantity of reducing sugar. A reliable method for characterising sprouted grains that can be used to predict the nutritional and compositional changes of oats during sprouting and, ultimately, for product development and specification, might be developed using the concept of degree of sprouting.
6.1.6 Changes in the Component After Sprouting 6.1.6.1 Protein Content Numerous factors that affect the metabolism of amino acids and proteins during germination and sprouting are temperature, oxygen, time, humidity, light, grain protein concentration, and growth regulators. The dry weight losses from respiration during malting may be the cause of the rise in proteins. As a result, compared to the un-germinated material. There would be more oat seeds per weight of the germinated seeds (Tian et al., 2010). The reason is that following germination, the proteins in the uncooked oat seeds were broken down and changed into a soluble condition. The concentration of free amino acids was reduced during steeping, mainly during the early stages of germination, which was a noteworthy finding. The bio enzymes may be produced from amino acids more quickly than proteins could be broken down into amino acids. 6.1.6.2 Phenolic Content The phenolic acid profile includes several free phenolic acids in oat sprouts and oat grains, such as protocatechuic acid, gallic acid, p-hydroxybenzoic acid, cinnamic acid, syringic acid sinapic acid, p-coumaric acid, caffeic acid and ferulic acid. Gallic acid and its derivatives hydroxybenzoic acid and syringic acid, made up the majority of the hydroxybenzoic acid profile. In both oat grain (OG) and oat sprouts (OS), p-hydroxybenzoic acidis predominated, followed by gallic acid and
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3,4-dihydroxybenzoic acid. As a result, these phenolic acids have been identified as important oat grain components. The germination of oats results in the loss of the gallic acid, hydroxybenzoic acids, ethyl ester and syringic acid. Depending on the genotype and environmental factors, oat sprouting has a different phytochemical content and metabolism. Ferulic acid derivatives, such as the isomers of feruloylquinic acid and diferuloylquinic acid, are higher in concentrations in the OS than in the OG. With variations in steeping and germination stages, phenolic compounds and antioxidant activity gets change. The phenolic content is not significantly affected by a brief steeping treatment. As a result of germination, the amount of free phenolic compounds is increased while the number of bound phenolics decreased. The number of free phenolic compounds increased while the number of bound phenolics decreased as a result of germination. 6.1.6.3 Flavonoid Content Seven minor flavonols were also considerably elevated by the germination process. Oat germination resulted in the loss of quercetin pentoside and kaempferol trihexoside, which were only found in the OG, indicating that they were catabolized during germination. Apigenin aglycone is not found, but nine flavones including luteolin and five variants as well as three apigenin derivatives were. The primary flavones in the OG are identified as apigenin hexoside and luteolin hexoside. Since these substances, together with apigenin malonyl-apiosyl-hexoside (F-16), are not present in the OS, it is interesting to note that the primary flavones of the OG get lost during germination. Additionally, after oat germination, luteolin rutinoside dramatically decreases. Cultivar genotypes, therefore, have a significant impact on the polyphenol profile. Throughout the germination stage, seven minor flavonols markedly increased. 6.1.6.4 Other Content Dark red chemicals can be produced when dinitrosalicylic acid reacts with reducing substances, particularly reducing phenolic compounds in oat seeds. Oat seeds’ primary chemical component, making up roughly 60% of their dry weight, is starch.
6.1.7 Benefits of Oat Sprouts Because sprouting has been proven to boost amounts of protein, phenolic compounds, essential amino acids, and γ-aminobutyric acid (GABA) during oat germination. Another benefit of sprouting is that the concentration of phytic acid is reduced. Phytic acid is frequently reduced in bread during the sourdough leavening
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process because it prevents mineral bioavailability. On the other hand, sprouting has effects that go beyond the good. Cell wall material, in particular glucan, a soluble fibre with health advantages, is removed during the sprouting process. Glucan causes an increase in intestinal viscosity, which slows down glucose absorption and lowers blood sugar levels as a result. The occurrence of microbiological activity during sprouting is another problem. Conditions that are typical for sprouting, like prolonged steeping times, high moisture contents, and temperatures as high as 25–30 °C, encourage the growth of microbes while also encouraging undesirable fermentation processes. As a result, growths of bacteria, mould, and yeast were found during sprouting, and it’s possible that latent spores were also reawakened. By regularly consuming sprouts, one can eliminate digestion issues, to make heart function normally, and improve the tone of your blood vessels. Additionally, sprouted oats reduce stress and promote happiness. Additionally, sprouts can enhance a woman’s appearance. Oats that have been sprouted are suitable for almost everyone and have numerous advantages. Only those who are allergic to gluten should refrain. Those who have digestive tract issues ought to be more cautious when eating the sprouts. Sprouted oats are a little sweeter and nuttier than unsprouted oat groats. Additionally, the texture is more sensitive. Oat groats release enzymes to break down their nutrients into building blocks when it’s time for germination so they can utilise these building blocks to grow. Proteins are broken down into amino acids and peptides, complex carbs are broken down into sugars, and fats are broken down into fatty acids. These more digestible molecules are smaller and simpler. There’s another explanation for why they might be simpler to process. Oats that have not been sprouted contain phytic acid, which might prevent stomach from producing digestive enzymes. However, this phytate is broken down by the germination process that starts in sprouted oats so that stomach can develop the digestive enzymes it needs to quickly digest the oats. It is well recognised that phytate reduces the body’s ability to absorb vitamins and minerals. The body can absorb more of the beneficial elements in sprouted oats, such as folate, iron, vitamin C, zinc, magnesium, and protein since the germination process breaks down this phytate.
6.1.8 Nutritional Value The increase in vitamins increase the nutritional content of sprouted oats, but the breakdown of beta-glucan is seen as a drawback. Therefore, balancing the quantities of these two nutrients is necessary to optimise the sprouting process for maximum nutritional benefits. Because of the physiological changes that occur during sprouting, the redox equilibrium is disturbed and stress is applied to the grain. Secondary metabolites, including vitamins and antioxidants like phenolics, are thus activated to enhance the sprouting. Figure 6.1 shows health benefits of the sprouted oats.
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Fig. 6.1 Health benefits of sprouted oats
6.2 Kamut Sprouts Kamut, Polish wheat, and Khorasan wheat are all varieties of the annual plant Triticum turgidum, sp. Turanicum, is in the same family as wheat, oat, and rye. This plant has a hollow, cane-shaped, knotted stem that can grow up to 1.3 m in height. The short, lanceolate, and glabrous leaves. A tall, slender spike with hermaphrodite- arranged flowers makes up the inflorescence. From August to September, when the seeds of this plant are ready to be gathered, it blooms from June to July. The portion meant for consumption is the grains or seeds. The kamut grain is larger than wheat (nearly double the size) and contains more fat. It is also golden in colour. About 25–25 grains are present in each ear. It has a delicious buttery flavour. In terms of nutrition, it is higher in lipids and minerals than wheat.
6.2.1 Composition of Kamut Sprouts Like other cereals, kamut is a grain high in complex carbohydrates. It is not a suitable cereal for celiacs because it contains gluten among its proteins. It has a more flavorful flavour since it has more fat than wheat grain. It is primarily composed of polyunsaturated fatty acids (60%) and Omega-6 linoleic acid. The whole grain makes a great contribution to fibre content. It contains insoluble fibre, which lowers cholesterol and helps avoid constipation. Regarding the vitamin and mineral content, this cereal stands out due to its higher contribution of vitamin E when compared to other kinds of cereals. The comparison of chemical composition of kamut and oats is shown in Table 6.4. The skin naturally produces vitamin E, which
166 Table 6.4 Chemical composition of Kamut (Khorasan wheat) and Oat
P. Kesarkar et al. Composition Water (%) Energy (kcal/100 g) Proteins (%) Carbohydrate (%) Total fibers (%) Sugars (%) Lipid (%)
Kamut (Khorasan wheat) 11.07 337 14.54 70.58 11.1 7.84 2.13
Oat 10.42 389 15.7 66 12.7 0.41 5–9
functions as an antioxidant and improves the appearance of the skin. Like all cereals, kamut is full of B vitamins, which support the body’s use of carbs and nourish the brain. Foods high in vitamin B, such as kamut, are advised for students, persons under stress, athletes, those with memory loss, and generally anyone who wants to focus and feel more energised. Selenium and zinc, antioxidant minerals that delay the onset of ageing, are abundant in this cereal. Regarding the vitamin and mineral content, this cereal stands out due to its higher contribution of vitamin E when compared to other kinds of cereals. The skin naturally produces vitamin E, which functions as an antioxidant and improves the appearance of the skin. Like all cereals, kamut is full of B vitamins, which support the body’s use of carbs and nourish the brain. Foods high in vitamin B, such as kamut, are advised for students, persons under stress, athletes, those with memory loss, and generally anyone who wants to focus and feel more energised. Selenium and zinc, antioxidant minerals that delay the onset of ageing, are abundant in this cereal. Kamut is a source of minerals selenium and manganese, which have an antioxidant effect. Selenium and manganese, which have an antioxidant effect, can be found in kamut. Water, organic materials, and minerals make up the grains. Nitrogen- free extractives (NFE), which are primarily starch and some sugars and range in content from 50% to 70% and even up to 75% in hulled rice, are the main components of organic substances. These are followed by crude proteins, which have a range of 8–18% and higher, and cellulose, which has a range of 2–11%. The dried grain has a water content of 14–15%. The majority of the nitrogen-free extractives present in grains are starch, which is located in the endosperm and accounts for around 80% of all carbs. The remaining portion of the grain’s weight is made up of sugars, mostly cane sugar, which is predominantly present in the grain’s germ. The germ has no starch. Between 2% and 4% of the grain in cereals is oil. Approximately 14% of wheat, 12.4% of rye, up to 26% of oats, and 20% of millet have the oil in their germ. The grain husk is the principal source of mineral ash. The majority of the minerals are removed during complicated grinding, and the finer the grind, the less mineral stuff is present in the flour. Roughly 50% of the ash in cereals is made up of phosphorus, followed by about 30% of potassium, 12% of magnesium, and just about 2.8% of calcium. Table 6.5 comprises mineral element composition of kamut flour and control wheat flour.
6 Oat and Kamut Sprouts Table 6.5 Mineral element composition of Kamut flour and Wheat flour
167 Variable (mg/kg) Potassium Magnesium Phosphorus Zinc Iron
Kamut flour 2663.81 916 3.47 24.97 24.17
Wheat flour 1546.53 513.16 0.93 15.1 20.01
6.2.2 Use in Human Nutrition Khorasan wheat has a high amount of carotenoids and a special nutraceutical value due to its unusual concentration of bioactive phytochemicals. Khorasan wheat also has a lot of selenium. Selenium levels in kamut khorasan bread were found to be ten times greater than those in contemporary durum bread, according to research. Numerous enzymes that protect cells from oxidative damage contain selenium in their active sites, including glutathione peroxidase and other selenoproteins.
6.2.3 Parameters Affecting the Nutritional Quality of Sprouted Kamut 6.2.3.1 Seed Source and Genotype The genotype is the most important factor in determining the nutritional value of sprouted grains. In the case of the winter and spring wheat types, the environment has also been identified as the primary factor causing the overall variance in some quality parameters. In the same area, years with lower temperatures and more rainfall in the 30 days before harvest had higher phenolic and flavonoid concentrations in durum and soft wheat. The mother plant’s susceptibility to biotic stress (such as diseases, weeds, and nutrient deficiency) and nutrient limitation also have an impact on the nutritional value of wheat grains. The protein content is typically lower in organic products, although older wheat types use nutrients more effectively in low- nitrogen conditions than modern wheat varieties, which are wholly reliant on high levels of available nitrogen. 6.2.3.2 Germination Condition During sprouting, biochemical changes depend on the germination circumstances as well as the “seed invigoration” treatments given to the grains to enhance germination and post-germination seedling growth. To produce sprouts of superior quality, particularly in terms of phytochemical content, research has recently focused on determining the ideal time and temperature combination during sprouting and pre- sowing treatments. Pre-sowing treatments don’t necessarily result in more bioactive
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chemicals building up in the soil. Brown rice and wheat both showed an increase in total phenolic compounds throughout the 24-h soaking period. Light Modulation One of the key elements influencing plant growth and development is light. To determine plant morphogenetic changes, the operation of the photosynthetic apparatus and the trend of metabolic pathways, photoreception systems respond to light intensity, quality, duration, and intermittency. Additionally, variations in lighting might cause photooxidative alterations in plants, which change how the antioxidant defence system functions. Uncertainty exists regarding the effects of a single wavelength under various lighting circumstances on the total phenol concentration, radical scavenging activity, and interactions with other antioxidants during seed germination. This is most likely because the amount of antioxidant molecules in the tissues is genetically defined, which has a big influence on seed sensitivity. Finally, it is important to note that, in addition to having an impact on the makeup of sprouts, light intensity, spectrum, and duration also have an impact on energy consumption and, consequently, the financial cost of indoor growing, which is the most typical scenario in sprout production. Temperature The GABA content rose in soft white winter and dark northern spring wheat during the first 48 h after germination and following successive hydrations with anaerobic and heat treatments (Benincasa et al., 2019). Salt Stress One of the most significant abiotic stresses on plants is salinity, particularly during the very salt-sensitive early stages of seedling growth. However, since the impact of salt stress has primarily been assessed in terms of germination rates and physiology, nothing is known about the influence of salinity on the phytochemical accumulation in edible sprouts yet.
6.2.4 Changes in the Component After Sprouting 6.2.4.1 Vitamin Kamut grain’s vitamin C concentration significantly increases during germination. However, when the germination period grew by an almost linear characteristic curve, the vitamin content progressively increase throughout sprouting and peaked
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at day 7 (around 550 mg/g). Both and tocopherol levels rise throughout the germination process compared to the ungerminated ones after steeping for 24 or 48 h. As the germination period lengthened during sprouting, the vitamin E content gradually increased (Bordoni et al., 2017). After 24 h of steeping, more vitamin E was generated than after 48 h. Wheat sprouts carotene concentration significantly rises during germination. Tocopherol was not discovered. The quantified vitamin E concentration is five times greater. The amount of vitamin C remained unaltered, however provitamin A is increased (Tian et al., 2010). The most powerful scavengers of free radicals are tocopherols. However, the availability of their regenerating systems, such as ascorbic acid, b-carotene, and selenium, as well as other criteria like their reactivity and concentrations, determine how active they are. On the other hand, glutathione and other cellular thiols are necessary for the regeneration of ascorbic acid from its radical state. 6.2.4.2 Phenolic Acids The majority of the wheat sprout’s antioxidants were phenolic acids. In the steeping water, it looked that free ferulic acid was being leached away and during the watering procedure, during the steeping phase and the first 4 days of germination. However, after 5 days, ferulic acid levels grew as a result of the hydrolysis of poly- phenolic substances linked to cell walls and phenolic biosynthesis. The substantial increase of these phenolic acids may potentially be caused by a significant drop in tannin content during the germination phase. As the majority of the free, during the cleaning and steeping processes, water-soluble phenolic acids gradually leached out of the mixture, the phenolic acid content often declined significantly over the first 4 days (Falcinelli et al., 2018). 6.2.4.3 Antioxidant Ascorbic acid, tocopherols, carotenoids, and phenolic substances are antioxidants that may be extremely important for seed germination and dormancy rupture. Antioxidants produced during germination are crucial for the safety of the young seedling because they slow down or stop the spread of free radicals produced during germination, which inhibits the oxidation of LDL and damages the cell membranes of young seedlings. The length of germination led to higher antioxidant concentrations. Antioxidant levels were typically marginally greater when grains were soaked for 24 h as opposed to 48 h before germination. In germinated wheat, phenolic chemicals made up the largest portion of the antioxidant content. There has been evidence of a synergistic impact between phenolic antioxidants, which can significantly boost the antioxidant activity of germinated kamut.
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6.2.5 Benefits of Kamut Sprout Although there is a very limited correlation between those results and the redox status that can be measured in vivo, several health advantages previously been claimed by invoking in vitro powerful antioxidant capability. Additionally, this method disregards additional biological effects that may be caused by substances that might not be identified by an in vitro assay but which, because of their high bioavailability, can deactivate or activate the oxidative metabolic pathways, affect other signalling pathways, or interfere with gene regulation, among other things. By using khorasan wheat flour instead of other ingredients, the resulting product’s nutritional value can be boosted. Bread made with Khorasan wheat flour had a greater protein level and more carotenoid content than loaves made with modern wheat. It has been established that kamut khorasan wheat may be used to make tortillas, bread, and cookies. After consuming kamut products, improvements were seen in several blood parameters, including minerals (magnesium, glucose, and potassium), inflammatory cytokines (IL-12, IL-6, vascular endothelial growth factor, and tumour necrosis factor-a), metabolic biomarkers (total cholesterol, low-density lipoprotein cholesterol, and alanine aminotransferase), and also redox status parameters (oxidised carbonyl proteins and lipid peroxidation) (Sofi et al., 2013). The nutritional content of the completed product could be increased by substituting Khorasan wheat flour for other ingredients. The weight of a thousand seeds (WTS) varies significantly amongst the types. The average WTS of the Khorasan variety is substantially higher than that of Kabo. The most significant feature of this actual grain, which sets it apart from the group of other species of the genus Triticum, is its quality, or the nutritional worth of its grains. Eating wheat sprouts will probably give the tissues access to a variety of highly concentrated antioxidants, such asb-carotene, vitamin C, vitamin E, and phenolic acids. Thus, kamut sprout may reduce plasma cholesterol (LDL cholesterol), scavenge free radicals from the body, and change immunological function in autoimmune disease patients by lowering inflammatory reactions. Sprouts of pseudocereal grains with high nutritional potential are now a great way to boost the impacts and advantages of nutrition. Raffinose and stachyose are removed during germination, and quantities of sugars and reducing starches are also greatly reduced. Due to a shortage of raffinose, which caused sucrose to be hydrolyzed for energy supply, the amount of sucrose may have decreased in the later stages of sprouting. 6.2.5.1 Food Safety Issues of Kamut and Oats Unfortunately, infections do become more prevalent in sprouts produced routinely. There is a significant increase in infections during the sprout-growing phase, according to numerous research. It goes without saying that when the seeds are planted, at least one disease must be present. Green sprouts are typically grown in a gently
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rotating drum at room temperature with “irrigation” water being added every 15–30 min. In addition to developing sprouts, this environment is ideal for growing diseases like Salmonella, E. coli, and even Listeria monocytogenes. Production of sprouts requires a high level of hygiene and quality control. The warm, humid conditions necessary for sprouting are also perfect for disease bacterial growth. Salmonella and E. coli O157: H7 bacteria will swiftly multiply in contaminated seeds to hazardous levels, resulting in foodborne disease. The Food and Drug Administration advises completely cooking sprouts for safety. Vulnerable groups, such as kids, expectant mothers, the elderly, or people with weakened immune systems, shouldn’t be given sprouts raw. Whether they are made at home or bought in a store, raw sprouts should be refrigerated. Sprouts can be consumed raw, but to reduce the risk of contracting a foodborne illness, thoroughly heat them to at least 135 °F before consuming. Sprouts are one of the top 10 foods that cause foodborne illness, and they have been connected to more than 40 outbreaks of foodborne illness between 1996 and 2016. Sprouts are frequently plucked and eaten right away since they naturally exhibit a quick quality deterioration at relatively low temperatures. To control the headspace of the package during its shelf life in terms of chemical composition, active packaging, temperature control, and changing environment are required to optimise the storage conditions. Due to the absence of a post-germination kill phase, sprout ingestion has been linked to numerous foodborne outbreaks. Even virtually little information about cereal and pseudo-cereal sprouts has been discovered, several studies have been undertaken on these subjects. Despite the genotypic heterogeneity of grains in the biochemical composition may require further procedure improvement, the technologies utilised on other species are based on common assumptions. There are numerous potential post-harvest and pre-harvest sources of pollutants, including germination media, seed material, soaking water, along with transport, storage and handling of seedlings, can be blamed for microbiological contamination of sprouts. Therefore, starting with seed treatments, the production process should be optimised.
References Aparicio-García, N., Martínez-Villaluenga, C., Frias, J., & Peñas, E. (2021). Production and characterization of a novel gluten-free fermented beverage based on sprouted oat flour. Foods, 10(1), 139. https://doi.org/10.3390/foods10010139 Benincasa, P., Falcinelli, B., Lutts, S., Stagnari, F., & Galieni, A. (2019). Sprouted grains: A comprehensive review. Nutrients, 11(2), 421. https://doi.org/10.3390/nu11020421 Bordoni, A., Danesi, F., Di Nunzio, M., Taccari, A., & Valli, V. (2017). Ancient wheat and health: A legend or the reality? A review on KAMUT khorasan wheat. Int J Food Sci Nutr, 68(3), 278–286. https://doi.org/10.1080/09637486.2016.1247434 Falcinelli, B., Calzuola, I., Gigliarelli, L., Torricelli, R., Polegri, L., Vizioli, V., Benincasa, P., & Marsili, V. (2018). Phenolic content and antioxidant activity of wholegrain breads from modern and old wheat (Triticum aestivum L.) cultivars and ancestors enriched with wheat sprout powder. Ital J Agron, 297–302. https://doi.org/10.4081/ija.2018.1220
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Holmback, J., Karlsson, A., & Arnoldsson, K. C. (2001). Characterization of N-acylphosphatidylethanolamine and acylphosphatidylglycerol in oats. Lipids, 36(2), 153–165. https://doi.org/10.1007/s11745-001-0702-z Krapf, J., Kandzia, F., Brühan, J., Walther, G., & Flöter, E. (2019). Sprouting of oats: A new approach to quantify compositional changes. Cereal Chem, 96(6), 994–1003. https://doi. org/10.1002/cche.10203 Lasztity, R. (1998). Oat grain—a wonderful reservoir of natural nutrients and biologically active substances. Food Reviews International, 14(1), 99–119. Rozan, P., Kuo, Y.-H., & Lambein, F. (2000). Free amino acids present in commercially available seedlings sold for human consumption. A potential hazard for consumers. J Agric Food Chem, 48(3), 716–723. https://doi.org/10.1021/jf990729v Sofi, F., Whittaker, A., Cesari, F., Gori, A. M., Fiorillo, C., Becatti, M., Marotti, I., Dinelli, G., Casini, A., Abbate, R., Gensini, G. F., & Benedettelli, S. (2013). Characterization of Khorasan wheat (Kamut) and impact of a replacement diet on cardiovascular risk factors: Cross- over dietary intervention study. Eur J Clin Nutr, 67(2), 190–195. https://doi.org/10.1038/ ejcn.2012.206 Tian, B., Xie, B., Shi, J., Wu, J., Cai, Y., Xu, T., Xue, S., & Deng, Q. (2010). Physicochemical changes of oat seeds during germination. Food Chem, 119(3), 1195–1200. https://doi. org/10.1016/j.foodchem.2009.08.035
Chapter 7
Sprouted Legumes: Biochemical Changes, Nutritional Impacts and Food Safety Concerns Josephine Ampofo and Lord Abbey
7.1 Introduction The modern-day consumer is becoming increasingly aware of the relationship between food biomarkers and development of diseases. With this increasing knowledge, the food industry is under constant pressure to supply consumers with foods that are not only rich in basic nutrients but also provide appreciable contents of health promoting compounds. Although animal-based foods such as milk, muscle foods (e.g., meat and fish) and their related products are excellent in nutritional quality, they are also associated with high cost, negative environmental effects, short shelf-lifeand sophisticated processing (Song et al., 2016). Thus, the popularity of plants as alternativegreen, nutrient-dense, and shelf-stable food systems for human nutrition. These advantages over animal-based foods have led to tremendous Preface: Research has shown the potential evidence of legumes for positive human nutrition. However, nutrient-density of legumes is limited by the presence of antinutritional factors such as tannins, phytates, trypsin inhibitors and oligosaccharides. Thus, the need for processing before consumption. Nevertheless, the choice of processing should not only limit the presence of antinutritional factors but also be sensitive to various nutrients and health promoting biomarkers. Among such nutrition-sensitive processing methods is sprouting, which involves the hydrolysis of a seed’s storage macromolecules into its simple bioavailable forms, as well as de novo synthesis of nutraceutical compounds such as phenolics. Thus, in-depth understanding of the parameters associated with sprouting and its consequent health benefits, will not only increase the market value of legume sprouts but also guide the food industry in making critical decisions for functional food applications. J. Ampofo (*) Department of Food Science and Technology, University of California, Davis, CA, USA e-mail: [email protected] L. Abbey Department of Plant, Food and Environmental Sciences, Dalhousie University, Truro, NS, Canada © The Author(s), under exclusive license to Springer Nature Switzerland AG 2023 I. Majid et al. (eds.), Advances in Plant Sprouts, https://doi.org/10.1007/978-3-031-40916-5_7
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demands for increased production of plant foods and development of novel processing techniques targeted for their conversion into edible, safe and healthy forms. Among plant-based foods are legumes, which are staple foods in diverse global cultures (Nkhata et al., 2018). Legumes belong to the family Leguminosae, consisting of plants that have pods with embedded seeds. Notable examples of legumes include dry bean (Phaseolus vulgaris), faba beans (Vicia faba), lentil (Lens culinaris), mung bean (Vigna radiata), chickpea (Cicer arietinum), soybean (Glycine max), peanut (Arachis hypogaea), dry pea (Pisum sativum), and lotus (Nelumbo nucifera) (Samtiya et al., 2020; Siddiq & Uebersax, 2013). Legumes exploited for food purposes only are sub-classified as pulses, whereas those exploited for oil purposes (i.e., soy and peanut) are not. Irrespective of their classification, legumes are consumed either as whole seeds or used as ingredients in different food preparations. Nutritional value of legumes is due to their appreciable concentrations of protein, dietary fiber, fats, minerals and vitamins. Besides basic nutrients, legumes also consist of secondary metabolites such as phenolic compounds. Phenolic compounds in legumes are reported to reduce risks of cardiovascular diseases, diabetes, hypertension, obesity, and some cancer types (Mamilla & Mishra, 2017; Jayathilake et al., 2018). Despite these positive health benefits, legumes are also made up of antinutritional factors (e.g., trypsin inhibitor, phytate and tannins), which are phytochemicals that interfere with digestibility of nutrientsthrough the formation of complexes with digestive enzymes andthereby, limiting their bioaccessibility and bioavailability (Hall et al., 2017). Also, the presence of oligosaccharides (e.g., raffinose and stachyose), have been shown to limit bioavailability of essential mineral elements (e.g., iron, calcium and phosphorus), as well as production ofintestinal gasleading to discomfort in consumers upon ingestion (Jayathilake et al., 2018). Another factor that limits food application of legumes is the presence of off-flavors and their long cooking time (Roland et al., 2017). Therefore, there is a need for appropriate legume processing to enhance their digestibility and nutritional value. Different processes such as dehulling, soaking, boiling, extrusion, sprouting and fermentation have been employed for legume processing (Patterson et al., 2017). Among these processing methods, sprouting or germination has gained popularity in the marketspace due to its capacity to enhancing nutritional properties (Ghavidel & Prakash, 2007; Roland et al., 2017). Sprouting is a simple and inexpensive process, requiring dry seeds, waterand sprouting containers. Sprouting commences with soaking of dry seeds to imbibe water and ends with the elongation of embryonic axis (Montemurro et al., 2019). Once the seeds absorb water and break dormancy, endogenous enzymes are triggered leading to the breakdown of complex storage molecules into simpler forms to nourish the growing embryo (Aloo et al., 2021). Also, during sprouting, the breakdown of structural cell walls leads to the release of bound phenolic compounds into free forms, thus increasing their phenolic concentrations. Due to these positive metabolic changes, legume sprouts can be classified as functional foods because they provideadditional health benefits besides basic nutrition. It can be argued that these positive health impacts of legume sprouts have accounted for the rising interest among researchers and industries as displayed in Fig. 7.1.
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Fig. 7.1 Bibliometric analysis on “legume sprouts and nutrition” from Web of Science database. The chart displays published research papers from January 1991 to June 2022
Emphatically, the efficiency of sprouting depends on cultivar type, seed viability and conditions of the sprouting process (Ampofo & Ngadi, 2020). Since sprouting does not require sunlight or soil, legume sprouts are a great natural resource for fresh vegetables, especially during hot summers and rainy periods when fresh vegetables are less available (Bains et al., 2014; Cauchon et al., 2017). Thereby, the aim of this chapter is to discuss the process of legume sprouting, biochemical changes and final health benefits in food applications. Also, measures on safety of legume sprouts to avoid foodborne outbreaks will be discussed in detail.
7.2 Legumes for Human Nutrition Legumes are nutrient-dense food crops rich in protein, carbohydrate, polyphenols and micronutrients e.g., folate, iron and potassium (Siddiq & Uebersax, 2013). Since ancient times, legumes have been part of the dietary patterns in diverse cultures due to their rich nutritional profile, as depicted in Table 7.1. Legumes are rich in complex carbohydrates such as dietary fiber that can be linked to satiation and protection against chronic diseases (Liu et al., 2015; Veronese et al., 2018). According to Thompson and Brick (2016), fiber intake of the American public is half its recommended intake. Although consumption of whole grains helps
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Table 7.1 Biochemical composition and health benefits of some selected legume species Plant specie Common bean (Phaseolus vulgaris L.)
Lentil (Lens culinaris L.)
Pea (Pisum sativum L.)
Chickpea (Cicer arietinum L.)
Cow pea (Vigna unguiculata L.) Soybean (Glycine max L.)
Major biochemical composition Fat: 0.8–1.5%; protein: 21–25%; carbohydrate: 60–65%; fiber: 3–7%; phenolics: 2.36–5.62 mg/g Protein: 20–31%; fiber: 20.44%; fat: 1%; carbohydrate: 63%; phenolics: 1.37–47.6 mg/g Carbohydrate: 3–10%; protein: 20–30%; fiber: 22.03%; fat: 0.5–3.5%; phenolics: 1.88–27.5 mg/g Carbohydrate: 60–70%; fiber: 13.9%; protein: 18–24%; fat: 3%; phenolics: 8.02–10.84 mg/g Protein: 23–32%; fiber: 2.5–32%; carbohydrate: 60%; fat: 1%; phenolics: 46.48–119.61 mg/g Protein: 36.3–47.0%; fiber: 27.25–30.53%; fat: 14.9–23.3%; phenolics: 2.10–2.31 mg/g
Health benefits Antioxidative, anti-diabetic, antimicrobial, antihypertensive, and antithrombotic
References Lee et al. (2018b, b), Bessada et al. (2019), Mojica et al. (2014) and Nergiz & Gökgöz (2007)
Antioxidative, inhibition of pro-inflammatory molecules, anti-cholesterol, anti- cardiovascular, antidiabetic, and anticancer Antihypertensive, antioxidative, anticancer, anti-cholesterol, and anti-cardiovascular
Roy et al. (2010), Zhang et al. (2018), Garcia-Mora et al. (2014) and Singh et al. (2017) Aluko et al. (2015), Burger & Zhang (2019), Roy et al. (2010) and Fahim et al. (2019) Roy et al. (2010), Gomez-Favela et al. (2017) and Ribeiro et al. (2017)
Antidiabetic, antioxidative, anti-cardiovascular, antiosteoporosis, antihypertensive, anticancer, and anti-cholesterol Anti-atherogenic, anticancer, antioxidative, antidiabetic, anti-inflammatory, and anti-cholesterol
Ha et al. (2010); Awika & Duodu, (2016) and Jayathilake et al. (2018) Antioxidative, antidiabetic Xu & Chang (2007), anti-inflammatory, anti- Ademiluyi & Oboh cholesterol, anti- (2013) and Lee et al. cardiovascular, and anticancer (2018b, b)
increase fiber intake than refined grains, legumes contain about three-fold more fiber than whole grains (Chen et al., 2016). Thus, legumes can be postulated as more efficient to meeting fiber needs than whole grains. Besides dietary fiber, legumes are also rich in protein associated with satiety and weight management (Melby et al., 2017). Interestingly, the ratio of fiber to protein in legumes makes them novel, compared to animal foods and other plant crops: (a) animal-based foods contain protein and not fiber; and (b) other alternative plant-based foods do not supply a rich duo of fiber and protein (Didinger & Thompson, 2021). Additionally, legumes are also rich supplies of polyphenols such as hydroxybenzoic acids, hydroxycinnamic acids, flavanols, flavonols and many more (Siddiq & Uebersax, 2013). Although, polyphenol concentrations in legumes are not as high compared to fruits and vegetables, consumption of legumes will contribute a long way to healthy diets by providing additional benefits against risks of disease development. Irrespective of their nutritional
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quality, public acceptance of legumes remains a challenge due to the presence of compounds classified as antinutrients e.g., trypsin inhibitors, lectins, tannins, phytate and many more. Therefore, the need for nutrition-sensitive processing such as sprouting capable of eliminating/reducing legume antinutrients without compromising their nutritional value (Azeke et al., 2011).
7.3 Process of Sprouting There is a need for deeper comprehension in order to produce high-quality legume sprouts because sprouting is a metabolic process involving hundreds of biomolecular and enzymatic activities. Basically, sprouting is characterized by two cycles: cycle 1involves soaking and rapid imbibition of waterinto dry seeds until cell matrices and molecules are fully hydrated; and cycle 2 includes limited water uptake by hydrated seeds but vigorous metabolic reactivation for embryonic growth and subsequent cell elongation (Logan et al., 2001). A schematic diagram simplifying stages of sprouting is represented in Fig. 7.2. To avoid contamination along sprouting, decontamination of seeds is normally carried out as the first step. Chemicals such as sodium hypochlorite and calcium hypochlorite are normally used to treat seeds prior to sprouting (Ding et al., 2013). Succeeding decontamination is soaking, which is normally done with distilled water
Raw seeds
Mung bean
Soybean
Common bean
Soaking stage (12-24 h; 20-23 °C)
48 72 96
Sprouting stages (24-120 h)
24
120
Fig. 7.2 A schematic diagram reflecting the process of sprouting in mung bean, soybean and common bean at different stages. (Adapted from Maleki & Razavi, 2021)
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or salt solution. The purpose of soaking is to allow dry seeds to imbibe water and break dormancy. However, care must be taken during soaking because of the possibility of microbial load at this stage. Thus, the need for decontamination prior to soaking. Apart from microbial load, excessive soaking has been shown to induce fermentation in the presence of microorganisms whereas insufficient soaking limits embryonic growth and subsequent yield of nutritional compounds (Idowu et al., 2020). Thus, for efficient soaking, factors such as soaking temperature, seed weight to water volume ratio and duration of soaking are highly important to elucidate. Majority of sprouted legumes are reported to have been soaked for 24 h at room temperature (i.e., 20–25 °C). During soaking, there is the migration of water from the micropyle into the cotyledon, and finally into scutellum for the embryo to initiate sprouting. Thus, for efficient sprouting it is recommended for legume seeds to gain moisture content of 35–45% during soaking (Gooding, 2009). Upon maximum hydration, there is the liberation of growth hormones such as abscisic acid, ethylene and gibberellic acid from embryo to the aleurone layer, resulting in the production of hydrolytic enzymes including lipases, carbohydrase and proteases into the endosperm (Ikram et al., 2021). Hydrolysis of storage proteins, carbohydrates and lipids by proteases, amylases and lipases respectively, provides the growing embryo with energy needed to drive metabolic processes along sprouting. In addition to moisture, other parameters such as temperature, light and duration are crucial for the sprouting process after soaking. Depending on legume cultivar and the purpose of end sprouts, the process is normally conducted between 10 and 25 °C for 3–5 days under darkness (Dove, 2010).
7.4 Factors Affecting Sprouting 7.4.1 Genetics and Seed Viability Nutritional value of legume seeds is critically influenced by its genetic make-up. Concentrations and availabilities of nutrients in seeds for embryonic nourishment is determined by the seed viability which is linked with genetics, pre- and post-harvest conditions, as well as geographical origin (Benincasa et al., 2019). Seed maturity is a key determining factor for optimum sprouting. Seeds will not sprout until their embedded embryo attains physiological maturity at the time of shedding. Immature seeds will not have enough hormones and storage nutrients required for optimum embryonic growth, thus prolonging sprouting time or failure to sprout (Benincasa et al., 2019). In other studies, discussed in the review of Benincasa et al. (2019), exposure of the mother plant to nutritional shortage and abiotic stress led to alterations in nutritional value of wheat grains. In addition to primary nutritional compounds, contents of secondary metabolites in seeds were influenced by both agricultural practices and environmental stressors (Aloisi et al., 2016). It should also be mentionedthat, seed viability after harvesting from mother plant ranges from
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few weeks to many years, although this can be limited if seeds are not stored under proper conditions.
7.4.2 Water Water is no doubt an important parameter for efficient sprouting. Excessive watering of seeds can lead to reduced oxygen levels, tissue breakdown and microbial contamination whereas underwatering delays or prevents seeds from sprouting (Kurbitzky & Ziburski, 1994). Therefore, it is recommended for seeds to be kept moist than left sitting in water during sprouting. Water absorption (imbibition) is mainly influenced by seed coat permeability, although solute concentration also plays a role. During imbibition, there’s an in-built pressure in the seed as it absorbs water, leading to cotyledon expansion. The increase in pressure can cause splitting of seed coatdepending on the water retention capacity and quantity of water available for seed hydration along the sprouting process. Also, among seed macromolecules, protein has been linked with water absorption during sprouting and thus, since legumes are higher in protein than cereals and grains (Nkhata et al., 2018), it may be postulated that legumes absorb more water during soaking and sprouting compared to the former.
7.4.3 Temperature The effect of temperature on legume sprouts depends on species and variety as well as seed source and age (Vozzo, 2002). The temperature at which seeds sprout is very crucial because it has high influence on metabolic processes. Breaking down of storage moleculesinto their simpler forms involve enzymes and precursors with optimized activities at specific temperatures. Research suggested that sproutingcan be limited either at extremely high or very low temperatures. Irrespective of specie type, optimum temperature for legume sprouts range between 20° and 30 °C (Ampofo et al., 2020). It should also be highlighted that, long incubation periodsmay be required for efficient sprouting at low temperatures.
7.4.4 Light/Darkness Although literature has reported some works on sprouting under light, majority of reported works on legumes are conducted under darkness. In aggregate, legume seeds can germinate under both darkness and sunlight with no or limited interference (Berkelaar, 2010).
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7.5 Biochemical Changes in Legumes During Sprouting Besides food safety, one of the key reasons for food processing is maintaining or improvingnutritional quality, if possible. Sprouting is characterized by many metabolic processes that induce changes in nutritional and quality attributes. As discussed earlier, breakdown of endogenous biomolecules leads to the formation of primary and secondary metabolites (Di Gioia et al., 2017; Gan et al., 2019). Thus, influencing the biochemical composition and nutritional value of sprouted legumes compared to their dry seeds, as shown in Table 7.2. However, the extent of biochemical changes will depend on legume cultivar type, geographical source of seeds, sprouting conditions and analytical techniques applied for quantification. Thereby, this section of the chapter will discuss biochemical changes in selected legume speciesalong the sprouting process.
Table 7.2 Impact of sprouting on biochemical composition of some selected legumes Plant specie Chickpea (Cicer arietinum L.)
Sprouting conditions Soaking: 0.05% lime water for 12–48 h; time: 3–5 days; room temperature; darkness Lentils (Lens Soaking: Distilled water for culinaris L.) and 6 h; time: 6 days; faba bean (Vicia temperature: 20 °C; faba L.) darkness Red kidney beans Soaking: Distilled water for (Phaseolus vulgaris 6 h; temperature; 22 °C; L.) time: 4 days; darkness Pulse seeds Soaking: 5.5–24 h in distilled water; time: 1–7 days; temperature: 20°–25 °C; darkness Peanut (Arachis Temperature: 30 °C; time: hypogaea L.) 9 days; darkness Lentil (Lens Soaking: 24 h in distilled culinaris L.) water; room temperature; time: 5 days; darkness Lentil (Lens Soaking: 24 h; temperature: culinaris L.) 30° and 40 °C; time: 5 days; darkness Soybean (Glycine Temperature: 27 °C; time: 2 max L.) and 6 days; darkness
Biochemical changes + protein carbohydrate, crude fiber, vitamin C and iron
References Laxmi et al. (2015)
− thiamine; + niacin and riboflavin
Prodanov et al. (1997)
− cyanide, tannin, Megat et al. polyphenols and phytic (2016) acid + phenolic compounds and Xu et al. (2019) antioxidant activity
+ trans-resveratrol and antioxidant activity + total phenolic content
+ ACE I activity at 40 °C
+ flavonols, flavanols, hydroxycinnamic and hydroxybenzoic acids
Li et al. (2020) Hernandez- Aguilar et al. (2020) Mamilla and Mishra (2017) Guzman-Ortiz et al. (2017)
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7.5.1 Carbohydrates During sprouting, amylases catalyze the breakdown of storage carbohydrates such as starch into simple sugars (i.e., glucose and maltose) and subsequently into either non-reducing sugars such as sucrose (Zhang et al., 2015) or improved digestibility and bioaccessibility. The rate of starch hydrolysis into simple sugars depends on sprouting conditions and the legume specie. Hydrolysis of starch into simple sugars is the main source of energy that drives metabolic processes during sprouting (Zhang et al., 2015). Research by Ghavidel and Prakash (2007) showed improvedin- vitrostarch digestibility after 24 h of sprouting by 53–82% in different legume species such as green gram, cowpea, lentil, and chickpea. Another study demonstrated increased total sugars in sprouted kidney bean, mung bean, soybean and peanuts by 14, 22, 19 and 26%, respectively, compared to their raw seeds (Megat et al., 2016). From this same study, sprouting enhanced the yield of total dietary fiber from 27% to 40% in peanuts, 29% to 32% in mung bean, 32% to 73% in soybean, and 37% to 60% in kidney bean. The authors attributed these changes to sprouting time. As confirmed by early researchers, maximum hydrolysis of starch occurs between 48 to 72 h when amylase activity is at peak (Nirmala et al., 2000; Tian et al., 2010). To support the effect of time and amylase activity during sprouting, Zhang et al. (2015) found insignificant increase in concentrations ofreducing sugars at the first 12 h stage of different legume sprouting. However, continues sprouting after 12 h caused a 20 times increased accumulation of reducing sugars, with the authors attributing this observation to increased α-amylase activity. According to Mbithi-Mwikya et al. (2000) early stages of sprouting is characterized by low levels of soluble sugars because of limited availabilities of endogenous α-amylase. With respect to structural carbohydrates, dietary fibers form a major group. Dietary fiber is divided into soluble and insoluble fibers. Insoluble fiber (e.gs., cellulose, hemicellulose and lignans) does not dissolve in water or gastrointestinal fluids, remaining unchanged as it moves through the digestive tract, whereas soluble fiber (e.gs., β-glucans and arabinoxylans) dissolves in water and gastrointestinal fluids into gel-like substances as it passes through the stomach and colon (Hübner et al., 2013). Literature has reported inconsistent works on impact of sprouting on legume dietary fiber due to variations in specie type, sprouting condition and fiber fraction. For instance, sprouting at33.7 °C, 12 h light and 12 h darkness for 171 h; enhanced soluble, insoluble and total dietary fibers in chickpea by approximately, 21, 13 and 13%, respectively, compared to their unsprouted chickpea seeds (Domínguez-Arispuro et al., 2018). Similarly, Megat et al. (2016) observed significant increment in dietary fibers with sprouted legumes (i.e., peanuts, soybeans, mung beans and kidney beans) compared to their unsprouted counterparts. According to Domínguez-Arispuro et al. (2018), sprouting can alter cell wall polysaccharides of legume seeds leading to cell wall biosynthesis and formation of new dietary fiber as further discussed by Laxmi et al. (2015). They explained that components of crude fiber such as cellulose, lignin, and hemicellulose increased significantly while the seeds were sprouting. Thus, sprouting can be an inexpensive
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approach to increasing the fiber content of foods such as legumes. Sprouting of peas and lupin elevated fiber content by 100 and 456%, respectively (Rumiyati et al., 2012). The capacity of sprouting to increase fiber levels is positive for nutrition, especially among diabetic and obese populace since fiber-dense foods are correlated with low-glucose release (Chinma et al., 2015). Fiber has also been shown to increase satiety through the formation of gels in the stomach, thus slowing down starch digestion and gastric emptying (Yu et al., 2014). Furthermore, because of the inability of salivary and pancreatic α-amylases to break down fiber after ingestion, fiber upon reaching the colon is fermented by endogenous colonic bacteria into secondary molecules such as short chain fatty acids (i.e., butyrate, acetate and propionate) which are proven to help regulate satiety (McNabney & Henagan, 2017).
7.5.2 Protein Literature available on legume protein changes along the sprouting process seem to represent an inconsistent trend, with some reporting higher contents and vice-versa. Works reporting increased protein levels of legume sproutscorrelate this trend to the loss of dry matter such as carbohydrates and fats for respiratory purposes (Jan et al., 2017; Ongol et al., 2013). However, protein reduction along legume sprouting may be due to their hydrolysis via protease activities as discussed by Nkhata et al. (2018). Thus, it may be postulated that the final yield of protein is dependent on the duo actions of biosynthesis and breakdown. However, it looks like the need for protein biosynthesis supersedes its breakdown because of the urgent need for nucleic and amino acids, as well as enzymes associated with sprout growth and development (Moongngarm & Saetung, 2010). Sprouting of cowpea, green gram, lentil and chickpea improved in-vitro protein digestibility within ranges of 14–18%, compared to their raw seeds (Ghavidel & Prakash, 2007). Rico et al. (2021) investigated the impact of sprouting conditions at 15°–27 °C; for 1–5 days in darkness on lentil protein content. From their study, optimized protein content was observed in sprouted lentils at 22 °C for days of sprouting.
7.5.3 Lipids Lipids are present in legume seeds as triacylglycerols. During sprouting, activities of endogenous lipases release esterified fatty acids from triacylglycerols. Afterwards, formed free fatty acids are first broken down via β-oxidation and glyoxylate cycles into sugars to fuel seedling development (Graham, 2008). The dynamics of fatty acids production in legumes is dependent on pre-sprouting treatments, lipase activity and levels of individual fatty acids present over sprouting time (Benincasa et al., 2019). Lipid content of legumes show moderate increasement at early stages of sprouting but show an opposite trend as sprouting progress for respiratory purposes
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(Traore et al., 2004). Change in lipid contents in different legume species (common bean, lentil, chickpea, and lupine) upon sprouting (temperature: 25 °C; humidity: 80%; time: 4 days; darkness) were investigated by Atudorei et al. (2021). The authors observed significant lipid reductions in sprouted chickpea, common bean, lentil and lupine by approximately, 12, 13, 17 and 26%, respectively, compared to their raw seeds. Capacity of sprouting to reduce lipid content is beneficial to the food industry because it suggests protection against rancidity for prolonged shelf-life.
7.5.4 Antinutritional Compounds and Micronutrients Antinutritional factors are compounds in foods that inhibit digestion and bioavailability of macro- and micro-nutrients (Benincasa et al., 2019). Notable examples of antinutritional factors include trypsin inhibitors, tannins, phytates, phenols and fiber (Gilani et al., 2005). Bioavailability of minerals in legumes upon sprouting is dependent on antinutrient type and content, activation rate of hydrolytic enzymes, degree of binding with minerals in the matrix, or synergistic interactions of these factors (Idris et al., 2007). Trypsin inhibitors interfere with activities of proteases such as trypsin involved with protein digestion and bioavailability. Trypsin inhibitor is a major nutritional gap for legume utilization because of its stability during processing, even at high thermal processing. Notwithstanding its resistance to thermal processing, sprouting has been shown to limit concentrations of trypsin inhibitor (Mbithi-Mwikya et al., 2000; Zhang et al., 2015). Variation of micronutrients (i.e., vitamins and minerals) in legume sprouts are dependent on specie, and soaking and sprouting conditions (Ikram et al., 2021). Phytate is the major storage form of phosphorus in mature legume seeds (Samtiya et al., 2020). However, phytate is considered as antinutrients because it can limit bioavailability of mineral elements such as phosphorus, zinc, calcium, iron, magnesium, manganese and copperdue to strong chelation affinity between phytate and cations of these elements (Samtiya et al., 2020; Azeke et al., 2011). Besides thechelation effect, insufficient amount of phytase (i.e., the enzyme responsible for hydrolyzing the phospho-monoester bonds of phytate into phosphoric acid and myo-inositiol) in the human intestinal tract limits the breakdown of phytate to release phosphorus for absorption (Azeke et al., 2011). However, sprouting has been reported to help improve bioavailability of phosphorus through the activation of endogenous phytase (Benincasa et al., 2019). Despite these reports, literature has also shown that recommended dietaryconcentrations of phytate impact positive health benefits such as antidiabetic, anticarcinogenic and antioxidative activities (Kumar et al., 2010). In a previous study, in-vitro bioavailability of minerals (calcium, iron and phosphorus), and contents of antinutrients (phytateand tannin) were evaluated in sprouted and unsprouted cowpeas, lentils and chickpeas (Ghavidel & Prakash, 2007). The authors observed significant increase in total crude protein, digestible protein, bioavailable minerals in all investigated legumes after sprouting.
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These observations were attributed to the significant reductions in phytate and tannin by 18–21% and 20–38% ranges among sprouted legumes, compared to their control seeds. Sprouting of legumes has also been shown to increase levels of vitamins such as tocopherols (α-, β-, and γ-tocopherols), riboflavin (vitamin B2), and total niacin (vitamin B3) through de novo synthesis (Kim et al., 2012). Lentils and faba beans sprouted at 20 °C for 6 days led to reductions in thiamine contents, whereas an increasing trend was observed for riboflavin and total niacin (Prodanov et al., 1997). In other studies, vitamin C (i.e., ascorbic acid) was found in higher levels with sprouted mung beanand chickpea (Guo et al., 2012; Laxmi et al., 2015). Accumulation of ascorbic acid during sprouting has been linked with increased presence of D-glucose as a precursor for D-glucuronolactone, which is then hydrolysed into L-gluconolactone for subsequent hydrolysis into L-ascorbic acid (Desai et al., 2010). However, the soaking stage of sprouting should be done carefully because of the potential leaching of minerals and water-soluble vitamins. Also, some vitamins are unstable upon exposure to light and temperatures above threshold levels. Hence, sprouting conditions should be selected with careful consideration to avoid loss of micronutrients.
7.5.5 Phenolic Compounds Legumes contain appreciable concentrations of phenolic compounds such as phenolic acids, flavonoids, tannins, quinones, coumarins, lignans and many others (Singh et al., 2017). Many studies have shown the significant role of reactive oxygen species (ROS) as signal molecules for metabolic processes during sprouting. As previously discussed by Carocho and Ferreira (2013), ROS originate from oxygen reduction to superoxide radical, hydrogen peroxide radical, hydroxyl radical and singlet oxygen. After imbibition of water, ROS are generated via routes such as electrolyte transport chain in mitochondria, electron transfer of photosynthesis in the chloroplast, lipid catabolism in glyoxysome, and many others (Bailly, 2004). Notwithstanding their signaling roles, the presence of ROS above threshold levels leads to the destruction of DNA, nucleic acids, proteins, carbohydrate, and lipids (Carocho & Ferreira, 2013). Thus, leading to the signaling of antioxidants to help protect cells from generated ROS via two mechanisms (1) activation of endogenous antioxidant enzymes such as catalase, superoxide dismutase and glutathione peroxidase; and (2) triggering of enzymes (e.g., phenylalanine ammonia-lyase and tyrosine ammonia-lyase) and precursors (e.g., phenylalanine and tyrosine) associated with the phenylpropanoid pathway. This the main biosynthetic pathway responsible for production of phenolic compounds (Ampofo et al., 2020). The presence of phenolic compounds in legume seeds are represented in both free and bound forms. Compared to free phenolic compounds, bound phenolics are the most prevalent in raw seeds and appear bounded to structural molecules such as lignins, hydrolyzable tannins, cellulose, and proteins (Engert et al., 2011). However,
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during sprouting an inverse trend has been reported. From literature, contents of total phenolic compounds increase upon sprouting, with free phenolics being dominant than bound phenolics (Alvarez-Jubete et al., 2010; Pal et al., 2016). This effect has been linked with activities of cellulases and pectinases that break down structural molecules associated with bound phenolics, thus liberating them into their forms (Ampofo et al., 2020). López-Amorós et al. (2006) investigated the effects of sprouting on phenolic composition of common bean. From their study, raw bean showed no levels of flavonoids but showed phenolic acids including protocatechuic acid (32.8–41.1 μg/100 g), HBA (32.3–36.1 μg/100 g), vanillic acid (90.9–97.9 μg/100 g), p-hydroxyphenylacetic acid (45.8–51.6 μg/100 g), trans-ferulic acid (342–366 μg/100 g), and cis-ferulic acid (74.1–79.1 μg/100 g). Upon 6 days of sprouting under darkness, although protocatechuic acid was not detected, other phenolic acids such as HBA (11.5–15.7 μg/100 g), p-hydroxybenzoic aldehyde (14.4–16.8 μg/100 g), vanillic acid (74.5–77.9 μg/100 g), vanillic aldehyde (6.4–7.6 μg/100 g), trans-CA (102–113 μg/100 g), trans-ferulic acid (49.8–54.0 μg/100 g) and cis-ferulic acid (24.3–26.5 μg/100 g) were observed in sprouted beans. Additionally, sprouting led to the presence of flavonoids such as quercetin-3-rhamnoside (297–311 μg/100 g), kaempferol-3-rutinoside (305–323 μg/100 g), and kaempferol-3-glucoside, unlike the raw bean which showed no flavonoids. It should also be discussed that the authors further investigated the effects of time and light on phenolic compositions of common bean, with their results concluding sprouting under darkness as the optimum condition for maximum accumulation of phenolic compounds. Mamilla and Mishra (2017) also reported increased total phenolic content in chickpea and red lentil from 0.75 to 1.1 g/kg and 0.8 to 1.82 g/kg at 30 °C and 40 °C, respectively. Similarly, Lin and Lai (2006) observed increased total flavonoid content with sprouting at 40 °C, although this varied depending on legume specie (i.e., 0.17–0.25 g/kg for red lentils and 0.19 to 0.32 g/kgfor kidney beans). However, the authors observed a 91% increased total flavonoid content with chickpea sprouts at 30 °C, compared to raw chickpea seeds. The authors attributed their observations to variations in temperature requirements specific for each legume specie during sprouting. For instance, phaseoloid-clad legumes (e.g., kidney bean) has been shown to require higher sprouting temperatures than homogalgena legumes (e.g., chickpea and lentil) (Mierziak et al., 2014). Aguilera et al. (2013) observed mixed phenolics results with sprouted lentils, compared to the findings of López-Amorós et al. (2006). In their study, sprouting at 24 h increased bound phenolic levels by 1.6-fold, whereasprolonging sprouting time up to day 3 reduced bound phenolics level by 87%, compared to raw lentil seeds. Furthermore, Gharachorloo et al. (2013) investigated sprouting effects on total phenolic content of lentils, with their findingsdemonstratinga 32% increase in total phenolics with sprouted lentils than their unsprouted forms. Similarly, Cevallos- Casals and Cisneros-Zevallos (2010) evaluated effects of soaking and 7-day sprouting on phenolic composition offaba bean. From their results, total phenolics showed increased contents in the order of 7-day sprouts>rawseeds>soaked seeds. In another study, flavonoids such as quercetin derivatives increased by 4% and 56% in sprouted
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dark bean, compared to their raw and boiled seeds (López et al., 2013). In a further interesting study, saponins, flavonols and isoflavones were evaluated in whole sprouts, cotyledons and seed coats of sprouted black beans by Guajardo-Flores et al. (2013). According to theirfindings, accumulation of saponins was higher in whole sprouts than in cotyledons and seed coats. Their results showed that, saponin contents increased by about two-fold in whole sprouts and cotyledons after day 1 of sprouting. They also observed flavonols and isoflavones to be associated with seed coats, although their concentrations were decreasing with prolongment of sprouting time. However, the authors found no significant changes in concentrations of aglycone compounds along the sprouting process, irrespective of fraction. Overall, the study of Guajardo-Flores et al. (2013) recommended sprouting up to day 1 as the optimum time for maximization of saponins and non-glycosylated flavonols in whole sprouts and seeds coats of black bean, respectively. Notwithstanding these positive results, some works have also reported reduced levels of total phenols in some legumesprouts, especially at the initial stages of sprouting. For instance, Khattak et al. (2007a, b) observed reduced levels of total phenolic content in chickpea with increasing sprouting time. In another study, Mamilla and Mishra (2017) reported reduced levels of total phenolic contents in sprouted kidney bean compared to its raw form. The authors attributed their observation to thicker seed coats which reduced water absorption for cell hydration and metabolic processes. Furthermore, Al-Numair et al. (2009) observed a 45% decreased total phenolic content in two cultivars of faba beanafter 6 days of sprouting, with this trend linked to (1) leaching effect of soaking prior to sprouting; where free phenolics are leached from the seed coat into leaching water; and (2) activities of polyphenol oxidase, the enzyme responsible for browning of cotyledons during sprouting by oxidizing phenolic compounds into quinones (Ampofo et al., 2020). Thus, based on the above discussed works, it is clear that phytochemical changes in legume sprouts are dependent on legume specie and sprouting conditions. Since many research works have demonstrated the ability of sprouting to enhance concentrations of nutritional and health-promoting compounds, development of optimum sprouting conditions specific for each legume specie is crucial for their applications as nutrient-dense foods.
7.6 Health Benefits of Legume Sprouts As discussed in previous sections of the chapter, sprouting leads to biochemical changes with enhanced bioaccessibility and bioavailability of micro- and macro- nutrients. Therefore, sprouted legumes can be postulated as naturalfoods with dense concentrations of varied nutritional and bioactive compounds required for healthy human functioning (Fig. 7.3). This section of the chapter will discuss some potential health benefits associated with consumption of legume sprouts.
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• High nutrient bioavailability • Antioxidative • Anticancer • Antiinflammatory • Antiviral • Antidiabetic
Fig. 7.3 A graphical depiction of human health benefits associated with consumption of legume sprouts
7.6.1 Antioxidative Properties The antioxidant activity of a food system is determined by different methods such as ferric reducing antioxidant power (FRAP), 1,1-diphenyl-2-picryl-hydrazyl antioxidant assay (DPPH), trolox equivalent antioxidant capacity (TEAC), or 2,2′-azino- bis-3-ethylbenzothiazoline-6-sulfonic acid radical scavenging assay (ABTS) (Aloo et al., 2021). The principle of antioxidant activity is the donation of electrons or hydrogen atoms to neutralise unstable free radicalsand thus, protecting the cell from oxidative stress (Ampofo & Abbey, 2022). Phytochemicals such as phenolics, vitamin C and peptides are associated with antioxidant capacities. In a previous study with lentils, Amarowicz and Pegg (2008) observed reduced antioxidant capacity at early stages of sprouting until day 4 where an opposite trend was observed. Similar to this study, Khang et al. (2016) evaluated the antioxidant capacity of phenolic extracts from soybean sprouts, with the authors reporting maximum antioxidant capacity and total phenolic compounds in sprouts compared to raw soybean seeds. Furthermore, isoflavones from soybean sprouts demonstrated higher TEAC and FRAP than their raw seeds (Guzmán-Ortiz et al., 2017). Also, phenolic extracts from 4-day mung bean sprouts showed a 31% increased antioxidant capacity, compared to raw mung bean seeds (Świeca & Gawlik-Dziki, 2015). Peanut sprouts were also observed to possess higher antioxidant activity than its control, due to the presence trans-resveratrol being six times higher in peanut sprouts than raw peanuts (Ding et al., 2017). However, some studies have also reported decreased antioxidant capacities with legume sprouts. For instance, Erba et al. (2019) investigated changes in antioxidant capacity in sprouted chickpeaand green pea. Spouting was conducted under darkness for 3 days at 22 °C and 90% relative humidity. From their study, sprouting reduced total antioxidant activity in chickpea and green pea by about 8% and 11%, compared to their respective raw seeds. The authors attributed this observation to increase in free radicals (mainly hydrogen peroxide as physiological mediators) at the onset of sprouting (Bailly et al., 2008; Wojtyla et al., 2016). Thus, available
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phenolic compounds and other antioxidants may have neutralized free radicals, resulting in a net decrease with antioxidant capacity as sprouting continued. The authors further subjected sprouted chickpea and green pea to soaking and cooking, where they observed total antioxidant reductions by 32% and 52%, respectively, compared to initial sprouted chickpeas, whereas soaked and cooked sprouted green pea presented reduced antioxidant activities by 46% and 66%, respectively, compared to their untreated sprouted counterparts.
7.6.2 Anti-inflammatory Effects Kujawska et al. (2016) investigated anti-inflammatory activity of soybean sprouts treated with ferrous sulfate in male and female Wistar rats. According to their results, activities of endogenous antioxidant enzymes such as superoxide dismutase, glutathione peroxidase and glutathione S-transferase were significantly enhanced in all Wistar rats fed with soybean sprouts. Based on their findings, the authors concluded that soybean sprouts elicited with ferrous sulfate can be postulated as novel food ingredients with human anti-inflammatory potential.
7.6.3 Antidiabetic Properties Sprouted legumes have presented strong evidence for better glycemic control. Diabetes is a metabolic disease characterized by increased levels of blood sugar, a condition termed as hyperglycemia. Occurrence of hyperglycemia is due to different factors such as limitations in insulin secretion and defects in insulin action, or sometimes a combination of both factors (Aloo et al., 2021). Thus, foods enriched in compounds that can inhibit activities of enzymes (i.e., amylases and glucosidases) responsible for hydrolysing carbohydrates into simple sugars will help manage or prevent risks of diabetes. With respect to this, various in-vitro and in-vivo studies have associated sprout phytochemicals with antidiabetic properties. In one study, fructose-loaded spontaneous hypertensive rats were fed with mung bean sprouts to elucidate its antidiabetic potential (Nakamura et al., 2016). According to the authors, mung bean sprouts significantly reduced heart rates, serum triglycerides and total serum cholesterol, compared to unsprouted mung beans. Besides carbohydrate-dense foods, another factor responsible for diabetes is obesity. Sprouts has been shown to be dense in fiber (Rumiyati et al., 2012; Yu et al., 2014), which is further related with delayed starch digestion and subsequent sugar release. Notwithstanding its effects on sugar release, fermentation of fiber in the colon leads to the production of short-chain fatty acids such as butyrate and acetate. These short-chain fatty acids have been reported in literature to initiate weight loss through their capacity of upregulating genes associated with fatty acid oxidation and lipolysis (Canfora et al., 2015; Rumberger et al., 2014).
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7.6.4 Anticancer Properties Research has shown that, exposure of human cells to chemicals and nanoparticles can lead to risks of cancer development. Since humans are continuously exposed to cancerous chemicals, there is the need for natural foods dense in molecules capable of protection against risks of cancer development. Anticancer activity of coumestrol extracts from soybean sprouts were investigated with cancer cells (Ling & Chang, 2017). From this study, coumestrol from soybean sprouts limited prostate cancer via PI3/Akt, ERK1/2 and JNK-MAPK cell signaling pathways.
7.6.5 Antiviral Activity With the recent COVID-19 pandemic, viral infections have become a major global cause of death. Besides COVID-19, other notable human viral infections include hepatitis B and C, influenza, and human immunodeficiency virus (HIV) (Neagu et al., 2018). According to Kormuth and Lakdawala (2020), constant application of vaccines and medications to treat viral infections has led to the development of drug-resistant viral strains. A further limitation of anti-viral drugs is the dose- limiting toxic effects of some antivirals among certain groups of the population (Neagu et al., 2018). To help bridge these gaps, researchers are constantly searching for natural antiviral agents such as plant sprouts. Identificationof sprouts phytochemicals capable of serving as antiviral agents will help protect the current and future generations from viral infections and drug-resistant viruses. Antiviral activity ofphenolic extracts from mung bean sprouts were investigated with respiratory syncytial virus (RSV) and Herpes Simplex virus-1 (HSV-1) via assays of virucidal activity, prophylactic activity, cytotoxicity and virus yield reductions (Hafidh et al., 2015). Results showed significant inhibitions of RSV and HSV-1 at 2.2 × 10 and 0.5 × 102, respectively.
7.7 Food Safety Issues Contamination of sprouts can be linked with both pre- and post- harvest factors such as seed origin, soaking medium, sprouting conditions, transportation and commercialization parameters. In-depth discussions of sprout safety practices are detailed by Yang et al. (2013) and Ding et al. (2013). As discussed in previous sections, sprouting of legumes leads to the breakdown of proteins, lipids and carbohydrates into simple molecules for increased bioaccessibility and bioavailability. It is well known that the presence of water and nutrients are favorable conditions for bacteria growth. Since sprouting expose seeds to moisture and bioavailable nutrients, sprouts have become a pivotal avenue for microbial contamination, especially if the process
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is not properly planned. Also, because most sprouts are consumed without major processing, there is a higher risk for contamination and adverse safety concerns such as food-borne outbreaks. For instance, the United States recorded major multistate outbreaks of E. coli from sprouts between 2010 and 2017 (Carstens et al., 2019). To optimize sprouting for improved safety, physical (e.g., ultrasonication, supercritical carbon dioxide, plasma, and photosensitization) and aqueous-chemical (e.g., electrolyte water, ozoneand chlorinated water) treatments are currently being applied in the food industry (Aloo et al., 2021). Chlorine concentrations in the range of 50–200 mg/Lisapplied in the food industry as seed disinfectants due to its reported wide range of antimicrobial activity (Dikici et al., 2015; Praeger et al., 2018). Recently, advanced applications of chlorine as dioxide gas are applied for microbial decontamination. The effectiveness of chlorine against microbial growth was demonstrated with mung bean sprouts, where artificially inoculated Salmonellain mung bean sprouts was significantly controlled by utilization of chlorine as a dioxide gas treatment than washing with aqueous chlorine (Prodduk et al., 2014). The effectiveness was due to the capacity of chlorine dioxide gas to perforate inaccessible cells on sprout surface, thus leading to Salmonellainactivation as discussed by the authors. Another innovative approach for microbial control is the utilization of organic acids such as acetic, malic and lactic acids. This approach is also gaining popularity as an alternative to chorine decontaminationbecause of the possible inactivation of chlorine compounds by organic materials present on fresh products; in addition to their capacity to form different carcinogenic organochlorine compounds. For instance, exposure of soybean sprouts to lactic acid (0, 1.5, 2 and 2.5%, v/v) combined with thermal treatment (20, 40 and 50 °C) for 3 min against Shiga toxin producing E. coli (i.e., O157: H7 and non-O157 serogroups such as O103, O111, O145 and O26) demonstrated significant reductions with all investigated stereotypes, especially with 2.5% lactic acid at 50 °C (Dikici et al., 2015). Beside these methods, the US Food and Drug Administrationhas recommended in-depth guidelines for safe growing, transportation, storage and retailing of sprouts (National Advisory Committee on Microbiological Criteria for Foods, 1999). Additionally, to ensure the supply of safe sprouts, members of the International Sprout Growers Association (ISGA) are admonished to strictly adhere to regulatory and manufacturing guidelines.
7.8 Conclusion Legumes are rich in nutrients and non-nutrients bioactive and functional compounds of remarkable human health benefit. However, the nutritional value of legumes is limited by the presence of antinutritional compounds such as trypsin inhibitor, tannin and phytate. Antinutritional compounds bind to nutrients in a food matrix, limiting bioaccessibility and bioavailability. Although thermal processing has been
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shown to reduce concentrations of antinutritional compounds, it also imposes negative effects on nutritional compounds. In this regard, sprouting is an innovative processing approach to limit legume antinutritional compounds without compromising theirnutritional qualities. Additionally, sprouting of legumes leads to the breakdown of complex macromolecules into simple bioavailable sugars, amino acids and fatty acidsin addition to producing bioactive compounds such as phenolics, and vitamin C. Hence, sprouts are well known functional foods. Because sprouting is an inexpensive process requiring only seeds, water and sprouting chamber, it can be postulated as a green process capable of producing sustainable natural fresh foods beyond basic nutrition. However, although legume sproutshaveexperienced vast advancements over the years, there still exist limited data in areas such as (1) optimization as a function of specie or cultivar for improved nutritional value; (2) applications for novel functional foods with in-vivo proof of concept; and (3) techno-functional behaviours in diverse food formulations. With these challenges addressed, the market value and utilization of legume sprouts beyond household production will gain dominance.
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Dr. Josephine Ampofo has a background in Food Technology. Her research focus is on green food processing for sustainable human nutrition. Dr. Josephine Ampofo completed her BSc (Hons) Biological Sciences and MSc. (Food Science and Technology) from Kwame Nkrumah University of Science and Technology, Ghana, after which she pursued her PhD in Food Science and Agricultural Chemistry from McGill University, Canada. She is currently a Postdoctoral Fellow at University of California Davis, Department of Food Science and Technology where she does research, and assist in writing grants, as well as helping mentor undergraduate students. Her current research activities include novel processing of aquatic-based plant proteins and post-harvest processing of tree nuts for improved quality and nutrition. She also has eight publications in well-recognized scientific journals . Research Interests: Nutrition-sensitive food processing, climate-smart food systems, plant-based proteins, nutraceutical compounds Dr. Lord Abbey has a background in Horticulture and Crop Science with a research focus on sustainable horticultural production systems for human health and well-being. Having completed his BSc (Hons) Agriculture from the University of Ghana, Dr. Abbey continued his studies in the UK, The Netherlands and Canada. He is currently an Associate Professor (tenured) at Dalhousie University Faculty of Agriculture where he teaches and supervises undergraduate and graduate students. His research program is Plant Nutrition and Physiology. He holds several federal and provincial governments awards and other grants and supervised about 40 highly qualified personnel. Some of his current research activities include exploration of natural amendments and pyrolytic products, tropical ethnic crops production in NS; haskap fertility; apple and pear health; aromatic and medicinal plants; integrated nutrient management systems under field and greenhouse production systems; and value-addition and alternative uses of compost and vermicompost. He has over 80 publications including in press articles to his credit. He is the Secretary for the Executive Board of the Canadian Society for Horticultural Science (CSHS), a Board member of the Nova Scotia Institute of Agrologists (NSIA) Council, and member of the International Society for Horticultural Science (ISHS) . Research Interests: Compost, greenhouse, soilless growing media, plant nutrition, plant physiology, plant metabolism, plant propagation, stress physiology, sustainable food systems Institutional Profile: https://www.dal.ca/faculty/agriculture/plant-f ood-e nv/ faculty-staff/our-faculty/lord-abbey.html
Chapter 8
Kidney Bean Sprouts and Lentil Sprouts K. C. Dileep, Kanchan Bhatt, Satish Kumar, Rakesh Sharma, Priyanka Rana, Monika Thakur, and Priyanka Suthar
8.1 Introduction Pulses play an important role in human nutrition, being high in protein, vitamins, minerals and dietary fiber they are regarded as the protective and body building foods. Pulses generally have an amino acid composition complementary to the major cereals thus also has a role in reducing the chances of the protein or specific amino acid deficiency. Lentil (Len culinaris Medik) is a very important pulse crop while its cultivation and consumption is steadily increasing around the globe. They are well recognized for their high protein content (i.e., 24–30%), carbohydrates (60–63%), calcium (126.28 mg/kg) and iron (9.45 mg/kg) content (Isah et al., 2018; Majeed et al., 2017). The red kidney beans (Phaseolus vulgaris L.) are also potent functional foods and have repute for their high content of the dietary fiber (4%), protein (29.1%), vitamin E and some major unsaturated fatty acids. Lentils and kidney beans also contains significant volumes of various bioactive compounds, including dietary fiber, protease inhibitors, lectins, saponins, phytosterols, and phenolic compounds which have several health promoting and disease preventing properties (Rebollo-Hernanz et al., 2020). Further, the nutritional and functional status of these crops can also be increased by germination, soaking, processing etc. One of the emerging areas in the utilization of the pulses for various functional benefits is by sprouting which represents the product of germination and are harvested before the development of leaves while their nutritional status is the highest (Miyahira & Antunes, 2021). Germination/sprouting leads to the catabolism and degradation of main macronutrients, such as carbohydrates, protein and fatty acids, accompanied with the increase of simple sugars, free amino acids and organic acids thus increasing their K. C. Dileep · K. Bhatt · S. Kumar · R. Sharma (*) · P. Rana · M. Thakur · P. Suthar Department of Food Science and Technology, Dr. Y. S. Parmar University of Horticulture and Forestry, Nauni, HP, India © The Author(s), under exclusive license to Springer Nature Switzerland AG 2023 I. Majid et al. (eds.), Advances in Plant Sprouts, https://doi.org/10.1007/978-3-031-40916-5_8
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nutritional and functional value. Additionally, the germination and sprouting has been reported to accumulate some secondary metabolites such as vitamin C and polyphenols glucosinolates, isoflavones etc. in the seeds for which the technology has been exploited to its fullest in the recent times (Marton et al., 2010). Melatonin and g-Amino butyric acid (GABA) are exclusively found in lentils and kidney beans in relatively higher quantities and their content has also been reported to increase significantly during germination and sprouting. In addition, the release of bioactive peptides with ACE-inhibitory activity may also occur during seed germination mitigating the risk of CVD and can also reduce the chances of high blood pressure (Gan et al., 2017). Germination and sprouting have also been reported effective in reducing the anti-nutritional factors like protease inhibitors, lectin, increases the digestibility, reduced content of resistant starch and flatulence causing compounds which can reduce the functional value of the crops (Fouad & Rehab, 2015; Winarsi et al., 2020). Sprouted grains are gaining popularity especially amongst the health conscious people which escalated especially after the systematic and advanced research studies revealed the presence of various bioactive compounds in the lentils and sprouted kidney beans along with their health benefits. Because of the above reasons, the consumption of sprouts and germinated seeds has observed a sharp upsurge in the last decade and has gained significant importance as potential “functional foods (Santos et al., 2020). Certain factors which should be prior considerations for effective sprouting should include light, watering, sprouting time, temperature and the relative humidity. Usually, sprouting takes place in dark at ambient temperature i.e., 20–30 °C (Gan et al., 2017). Sprouts can be cultivated as both industrial conditions using equipment like rotary drums, various chamber/container or sprouting tray and at home using commercial sprouting kit, jars or container. The nutritional and functional quality of the obtained sprouts is affected by growing method, solution used for soaking and watering, temperature, availability of light and duration of sprouting. These factors further shall vary from species to species. These factors are the fundamental characteristics for the growth, structure and chemical composition of plant (Fiutak et al., 2019). The popularity of sprouts amongst the masses is mainly attributed to their rich nutritional profile, minimum processing, high concentration of certain essential nutrients like vitamins and minerals and reduced volumes of the anti-nutritional factors which in turn enhances the minerals bioavailability of the sprouted grains (Miyahira et al., 2021). The chance of food borne outbreaks in the sprouted grains is a challenging job for the food industry which can be a major determinant for potential scale-up the sprouts production industry. Therefore, appropriate strategies and well optimized germination parameters must be considered to eliminate or inhibit the pathogens present in sprouts. This chapter aims to provide an updated knowledge on the production, nutrition, anti nutritional factors, functional properties etc. of the lentils and red kidney beans to justify the enhanced consumption of their sprouts with projection for various functional properties (Mir et al., 2021). Figure 8.1 depicts the nutritional and nutraceutical health benefits of kidney beans and lentil sprouts.
Zinc
· · · ·
OTHER BENEFITS Reduces antinutrients Minimally processed Improved digestibility Economical method
IMPROVEMENT STRATEGIES · Bio-fortification · Genetic enhancement · Conventional and molecular approaches · Value addition
Fig. 8.1 Nutritional and nutraceutical benefits of kidney beans and lentil sprouts
Immune booster, anti-inflammatory, anti-oxidative, anti-viral, prevent diarrhoea and phenomena.
Phosphorus
Calcium
Iron
Vitamin E
B Vitamins
Vitamin C
Nutrients
Regulate protein activity and acid base balance, absorption and transportation of nutrients, bone mineralization.,
Essential for bone formation and growth, cofactor and regulator for biochemical reactions
Transport and storage of oxygen, cofactor for enzymes and proteins, formation of red blood cells
Prevents the oxidation of various molecules. Increase synthesis of haem, proper synthesis of nucleic acids
Involved in energy metabolism, Prevents beriberi, pellagra, ariboflavinosis, anaemia.
Prevent scurvy, anti-oxidative, increase iron absorption, improve immune system, detoxify drugs
Nutraceuticals
Health effects
Carotenoids
Biopeptides
Saponins
Phytosterol
Phenols
GABA
Melatonin
Immunomodulatory, reduce oxidative stress, cognitive protectants, anti-obese and anti-cancerous
ACE-inhibitor and anti-hypertensive, anti-oxidative, anti-inflammatory, anticancerous
Reduce cholesterol levels, anti-oxidative in nature, prevent inflammation, protect against nature
Reduce cholesterol levels in body and cholesterol absorption, antiinflammatory, anti-tumorous
Reduce blood pressure, protective against oxidative stress, Chemo-preventive, anti-diabetic in nature
Regulate blood pressure and heart rate, relieve pain and anxiety, increase the secretion of insulin, prevent cancer
Inhibit lipid peroxidation, reduce anxiety and depression, prevent inflammation and cancer, regulate sleep and anxiety.
Health effects
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8.2 Enhanced Nutritional and Functional Benefits of Sprouts The rapid increase in chronic health diseases and higher health awareness among the masses has resulted in a global increase in the demand for healthy food items. The food industry has always evolved to address the novel and continuously changing demands of the health conscious people for better and cheaper healthy food options. Lentils and kidney beans are abundantly available pulse crops which contain significant proportions of various nutrients and bioactive compounds, particularly phenolic acids, flavonoids, carotenoids, melatonin, GABA and phytosterols. These functional compounds are known to possess various immuno-modulatory, anti-oxidative, anti obese, anti diabetic and anti cancerous properties which are likely to be helpful to mankind as they cure under laying health issues (Rebollo- Hernanz et al., 2020; Wojdyło et al., 2020; Lee et al., 2022a, b; Mustafa et al., 2022). Sprouting is not a new technology however; its importance for increasing the functionality of the grains and improving the nutritional composition has recently been recognized. The trypsin inhibitors, which are present in many grains, posses a significant nutritional issue as they can interfere with the digestion process of the proteins in the body. They inhibit digestion because cooking cannot deactivate these inhibitors as they are heat resistant and stable at high temperatures (Nkhata et al., 2018). Another, important anti-nutritional factor present in lentils and kidney beans is phytate, a well known metal chelator. Despite having prohealth effects such as anti-carcinogenic, anti-diabetic, and anti-oxidant properties, they are characterized as anti-nutritional factors since they can form insoluble complexes with minerals such as potassium, iron, zinc, calcium, magnesium, copper, and manganese (Kumar et al., 2010). The sprouting process can, however, be helpful to reduce the concentration of both these anti-nutritional factors in the lentils and kidney beans (Kumar et al., 2010; Zhang et al., 2015). In addition to this, sprouting reactivates grain metabolism and leads to degradation of anti-nutrients and also increases the availability of the macronutrients. These compounds are then converted to secondary metabolites that have potential health benefits and important health modulation properties (Peñas & Martínez-Villaluenga, 2020). Raffinose is a broad family which contains the anti-nutrients like raffinose, stachyose, ajugose, and verbascose which can cause flatulence. Absence of β-galactosidase in human intestine causes their fermentation by the gut microbes, resulting in the formation of gases (carbon dioxide, hydrogen, and methane) which ultimately leads to flatulence which can be accompanied by abdominal pain, nausea, diarrhea, cramps and discomfort (Sharma, 2021). The bitter taste of lentils and kidney beans is due to the presence of various glycosides saponins and sapogenins, which are reduced during germination and are ultimately converted to simple sugar (Winrasi et al., 2020). After germination a significant decrease was observed in the content of TIA (Trypsin inhibitors activity) of white beans (52.5%) and black beans (25%) while, phytic acid was reduced by more than 40%, tannins by 19% and 36.2% in black beans and white beans respectively (Sangronis & Machado, 2007). Similarly, Ghavidel and Prakash (2007) reported that phytic acid and tannins were reduced by 18–21% and 20–38%, in the germinated green grass, cowpeas, lentils, and
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chickpeas. The effect of sprouting on the chemical composition of lentils and kidney beans has been summarized in Table 8.1. Additionally, sprouting is an excellent technique to improve the nutritional profile of most of the seed crops. It increases the digestibility of starch and protein in the seed as well as improves quality content of vitamins and amino acids and certain other biological compounds with significant functional properties. Breakdown of proteins and carbohydrates develops a delicious taste, color and flavor in the food product which at the same time also increase the efficiency scores of the food along with the bioavailability of the micro and macronutrients in the food (Mikulinich & Guzikova, 2021). Hormones and enzymes that hydrolyze food reserves, transport soluble food and hormones to the growth point, and promote assimilation of the secreted compounds during the germination of red kidney beans. Hydrolases (protease, lipase, and carbohydrates) are also activated during sprouting of most of the seeds (Chaudhary et al., 2013). Red kidney bean sprouts therefore contain more amino acids, fatty acids, and glucose than the un-sprouted red kidney beans. Sprouting in specific has been reported to increase the volume of vitamins such as folate, thiamine, pyridoxine, vitamin C and vitamin E in different germinated green bean, buckwheat, chickpeas, lupins, mung beans, and soybeans (Miyahira et al., 2021). The raw/unsprouted seeds of buckwheat, soybean and mung beans does not contain L-ascorbic acid however, its level, bioactivity and bioavailability elevates significantly during sprouting (Wojdyło et al., 2020). Sprouting can increases the protein digestibility by 2–4%, thiamine content by more than 26.7% and ascorbic acid by 300% and 33% for white beans and black beans respectively. A study on different lentil varieties depicted a significant increase in the concentration of protein after germination. Furthermore, the germination process also alters the micro- mineral levels, especially Zn and Mn and Ca (Santos et al., 2020). Protein, thiamin, in vitro iron, calcium, as well as in vitro starch and digestible protein content can significantly increase as a result of germination of green grass, cowpeas, lentils, and chickpeas (Ghavidel & Prakash, 2007). Sprouts are considered a rich source of various secondary metabolites (mainly phenolic compounds, phytosterols and glucosinolates) along with macro and micro-nutrients which are produced by aerobic respiration and biochemical metabolism. Especially, kidney bean sprouts and lentil sprouts contain higher amounts of melatonin and γ-amino butyric acid (GABA). In comparison with commercial adult plants, the germinated or sprouted seeds contain two to ten folds higher phyto-chemicals while their bioactivity is also enhanced due to these technologies. These contents depend on various factors such as environmental conditions, species, duration of germination, cultivars, processing and storage (Choe et al., 2018). The bioactive compounds present in kidney bean sprouts and lentil sprouts (KBS and LS) and their health modulation roles are explained below as well as represented category wise in Table 8.2. This is quite clear that germination and sprouting can be helpful technologies for increasing the nutritional and functional profile of most of the seed crops including the lentils and kidney beans. Various studies and biochemical profiling of the sprouted grains also supports the hypothesis of the enhanced nutritional profile of the grains which has led to certain upsurge in their consumption by the masses which is still expected to increase in the years to come.
61.13–91.8
40.76
118.5–233.04
Total polyphenols (mg GA/100 g)
Antioxidant activity (% DPPH scavenging activity) Phytic acid (mg/100 g)
Ascorbic acid (mg/100 g)
33.3
1341.13–1495.0 1510.10–1833.0 2400.00
62.19
2.77–4.16
Ash (%)
Trypsin inhibitor (TIU/mg protein)
14.90
10.70
Moisture (%)
156.0–466.10
41.69–53.85
Carbohydrates (%) 48.70–56.53
Tannins (mg/100 g)
6.75–25.40
8.11–21.70
27.37
131.0–189.20
4.97–6.97
3.30–9.14
4.13–4.37
1.7–6.1
6.1–9.11
45.5
3.8–6.6
10.9
68.2–77.4
2.4
0.3–4.2
Crude fiber (%)
0.75–0.93
1.1–2.20
17.7–21.6
Crude fat (%)
23.51–28.86
24.6–31.41
Proteins (%)
12.12–13.63
1.9–3.22
1.1–1.9
3.5–4.32
55.9
3300.00
4.3–6.8
12.0
60.8–78.2
5.1
0.1–3.3
15.7–17.8
Due to the enzyme activities of the seeds during the early stage of germination
Attributed to leaching and/or binding with substances i.e. carbohydrate or protein and also hydrolysis by polyphenolase
Apparently as a result of a large increase in phytase activity. Phosphates and inositol, which can both be generated by the hydrolysis
Dry legumes absorb water rapidly, influenced by the structure of the legume. Also, due to the increasing number of cells within the seed becoming hydrated These increases could be due to an increase in phytase activity during germination. Due to biosynthesis and bioaccumulation of phenolic compounds. Also, due to degradation of polymerized polyphenols, specifically hydrolysable tannins, and the hydrolysis of other glycosylated flavonoids Mainly attributed to an increase in the activity of the endogenous hydrolytic enzymes, polyphenols and also due to the synthesis of compounds like vitamin C and tocopherols
The alpha amylase breaks down complex carbohydrates to simpler and more absorbable sugars which are utilized by the growing seedlings
Attributed to the fact that part of the seed fiber may be solubilized enzymatically during seed germination
Germinated seeds exhibit increased proteolytic activity, which results in hydrolysis of storage proteins Attributed to the increased activity of lipolytic enzymes, which hydrolyzed the fats into fatty acid and glycerol for energy
Table 8.1 Effect of sprouting on the chemical composition of lentils and kidney beans Lentils Kidney beans Nutrients Unsprouted Sprouted Unsprouted Sprouted Justification Fouad and Rehab (2015), El-Adawy et al. (2003), HernandezAguilar et al. (2020), OskaybaşEmlek et al. (2021), Sangronis and Machado (2007), Azra Yasmin et al. (2008), De la RosaMillán (2019), GuajardoFlores et al. (2013) and Duenas et al. (2016)
References
K. C. Dileep et al.
Phytochemicals Specific functional compounds category KBS LS Hydroxycinnamic Phenolic acids Ferulic acid p-Coumaric acid acids Sinapinic acid Sinapinic acid b- resorcylic acid Galic acid Chlorogenic acid Vanillic acid p-Hydroxybenzoic Ferulic acid Rosmarinic acid acid Protocatechuic acid Quinic acid Procyanidins Flavonoids Catechin Catechins Eriodictyol Quercetin Hesperetin Isorhamnetin Naringenin Apigenin Quercetin Prodelphinidin Kaempferol Gallate procyanidins Isorhamnetin Luteolin Pelargonidin Cyanidin Malvidin Genistein Biochanin Myricetin Mechanism of action in the body Anti-oxidative: Different phenolic compounds act as antioxidants and have ability to reduce formation of ROS i.e. superoxide anion, by chelation or enzyme inhibition. Anti-diabetic: Good inhibitors of α-glucosidase and lipase and so contribute significantly to the control of blood sugar and obesity. Cardioprotective: Reduce hypertension by inhibiting the angiotensin I-converting enzyme along with antihyperlipidemic, hypohomocysteinemic, anti-cholesterolemic. Anti-cancerous: Chemo-preventive activities include the uptake of carcinogens, activation or formation, detoxification, binding to DNA and fidelity of DNA repair.
Table 8.2 Bioactive compounds present in kidney bean sprouts and lentil sprouts and their health benefits
(continued)
References Rebollo-Hernanz et al. (2020), Hernandez-Aguilar et al. (2020) and Dueñas et al. (2015)
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Melatonin
–
–
Phytochemicals Specific functional compounds category KBS LS Carotenoids – Neochrome Neoxanthin Zeoxanthin Lutein Violaxanthin Carotene
Table 8.2 (continued) Mechanism of action in the body Immunomodulatory: Reduces the serum levels of interleukin 8 (IL-8), tumor necrosis factor α (TNFα), interleukin-6 (IL-6) and C-reactive protein (CRP) in the overweight and obese cohort. Anti-obese: Increase adiponectin and decrease monocyte chemotactic protein 1 (MCP-1), with a concurrent significant decrease in body weight. Anti-oxidative: quenches single-oxygen and inhibit the oxidation of LDL, prevent plaque formation and coronary artery disease. Cognitive: long-term β-carotene supplementation (50 mg on alternate days) helped to maintain cognitive performance it may due to antioxidant activity Immunomodulatory: decreases TNF-α, interleukin (IL)-1β and IL-6, myeloperoxidase, and malondialdehyde levels and increased glutathione and superoxide dismutase levels Sleep and anxiety: increased sleep efficiency, immobile time, and decreased total nocturnal activity, sleep fragmentation index, and sleep latency. Also, improvement in anxiety and depression Anti-oxidative: quench free radicles, chelate metals and acts synergistically with other antioxidants. Also, enhance the activity and gene expression of antioxidant enzymes (glutathione peroxidase, glutathione reductase, and superoxide dismutase). Anti-cancerous: by stimulating apoptosis, anti-growth signaling and by inhibiting proliferation, angiogenesis, immune evasion.
Rebollo-Hernanz et al. (2020) and Gonçalve et al. (2021)
References Wojdyło et al. (2020) and Eggersdorfer et al. (2018)
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Phytosterols
β-Sitosterol β-Sitosteryl Stigmasterol Campesterol
β-sitosterol, camppesterol stigmasterol
Phytochemicals Specific functional compounds category KBS LS γ-aminobutyric – – acid (GABA) Mechanism of action in the body Anti-depressive: increased neurotransmitters including dopamine, 5-hydroxytrptophan (5-HTP) and norepinephrine in hippocampus of mice, which promoted social communication and pleasure resulted from physical movements. Anti-diabetic: stimulate the proliferation of β-cell, which enhanced production of human insulin. Anti-oxidative: scavenge free radicals, captures the reactive carbonyl compounds such as malondialdehyde formed during lipid peroxidation. Anti-cancerous: decrease the regulation and production of MMP-2 and MMP-9, which are the proteinases that induce penetration of tumor cells in cholangiocarcinoma QBC939 cancer cells. Anti-hypertensive: Greatly inhibited the activity of angiotensin-I converting enzyme that associated with blood pressure rising. Anti-cholesterol: compete with cholesterol, decreasing its micellar solubility in the intestine and reduces cholesterol absorption. Also decrease the biosynthesis of cholesterol by reducing the expression of the HMGCS1 gene. Anti-cancerous: PS are reported to prevent and protect against common cancer types such as prostate, breast, colon, and other cancers (continued)
Ramırez-Jimenez et al. (2015), and Mustafa et al. (2022)
References Zhang et al. (2018) and Lee et al. (2022a, b)
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Organic acids
Oxalic acid Citric acid Malic acid Succinic acid
Phytochemicals Specific functional compounds category KBS LS Soyasaponin VI Saponins Soyasaponin soyasaponin βg Deacetyl Soyasapogenol B soyasaponin
Table 8.2 (continued) Mechanism of action in the body Anti-oxidative: capture free radicals and also activate antioxidant enzymes. It binds cholesterol and prevents its oxidation in the colon. Anti-cholesterol: Form an insoluble saponin–cholesterol complex, which inhibits cholesterol absorption and increases fecal excretion of bile acids. Anti-inflammatory: inhibit the production of pro-inflammatory cytokines (TNF-α and MCP-1, PGE2 and NO), inflammatory enzymes (COX-2 and iNOS) and the degradation of IκB-α (an inhibitor of NF-κB, in LPS-stimulated macrophages) Anti-cancerous: Can interact with free- or membrane-bound sterols in the colon and stimulate their fecal elimination, which is correlated with cancer prevention. Anti-diabetic: promote satiety and delay the gastric emptying time, thereby decreasing the postprandial glucose and insulin response Anti-oxidative: organic acid acts as cheating agents of metal ion and act as antioxidants indirectly. Immunomodulatory: Lactate act through the regulation of the cellular cytokine network and the signaling systems of the intestinal mucosal system Anti-cholesterolemic: Serum cholesterol and triglycerides were reduced by 0.3% acetic acid in rats.
Wojdyło et al. (2020) and Shi et al. (2022)
References Zhang et al. (2018) and Guajardo-Flores et al. (2013)
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8.2.1 Antioxidant and Anti-inflammatory Activity Antioxidants are the molecules which combat against various oxidative stress conditions by preventing the oxidation of other molecules (Gulcin, 2020). Kidney bean and lentil sprouts have ample of phyto-nutrients like phenols, flavonoids, melatonin, carotenoids, glucosinolates, tocopherols and γ-aminobutyric acid (GABA) etc. which have reported potent antioxidant properties. These compounds have several different mechanisms to reduce the oxidation of various cell molecules, which include free radicals quenching, chelating metals, acts synergistically with other antioxidants and enhancing the activity of antioxidant enzymes. Phenolics which are present in abundance in the sprouts are regarded as main compounds responsible for the antioxidant activity of the lentils and the sprouted beans. Since, melatonin and GABA are two major antioxidants in the lentils and the kidney beans and the researchers have recently been concentrating on the significance of these substances in preventing a variety of lifestyle related disorders (Zhang et al., 2017a, b; Lee et al., 2022a, b). Melatonin (1.01 g/g) and phenolic compounds (3.47 mg/g) were reported to have the highest concentrations in sprouted lentils after a 6 days germination period. Compared to raw lentils, sprouted lentils have a higher melatonin concentration (more than 2000 times higher) thus the protective role against various degenerative health diseases is also enhanced after the sprouting. Owing to the changes produced by germination, the antioxidant capacity of the germinated lentils was higher (3.8-fold) in comparison to that of raw lentils. In another study Reiter et al. (2005) reported a reduced oxidative stress and an increase in plasmatic melatonin levels in the germinated kidney beans while, Aguilera et al. (2016), found that after administration of the kidney bean extract to rats, the plasmatic melatonin levels were increased which in-turns reduced the level of various oxidants in the plasma. In other studies, the protective effect of GABA against H2O2-induced oxidative stress in pancreatic cells (Tang et al., 2018) and human umbilical vein endothelial cells (Zhu et al., 2019) was observed via reduction in cell death, inhibition of reactive oxygen species (ROS) production, and amplification of antioxidant defense systems. Saponins are yet other class of antioxidants that are gaining popularity because of their capacity to bind to cholesterol and prevent its oxidation in the colon. Saponins can remove free radicals or can activate antioxidant enzymes to prevent their interaction with the free radicals with metallic ions (Luo et al., 2016). Several studies have also supported the fact that saponins from lentils may also act as anti-inflammatory agents. The legume based diet (lentils and other legume) effect on elderly people was recently studied by Conti et al. (2021), especially for their anti-inflammatory properties and they reported reduced levels of various inflammatory enzymes such as COX-2 and iNOS and pro-inflammatory cytokines like TNFand MCP-1, PEG and NO. Certainly, the antioxidant properties of the beans and lentils can be enhanced by the sprouting technology to make them healthier options for the health seekers. This has resulted in the percolation of the luxury sprouted seeds from the five star hotels to the plates of the health conscious people which would surely sustain the health and reduce the pressure on the healthcare systems as well.
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8.2.2 Anti-diabetic and Anti-obese Diabetes is a chronic metabolic disorder characterized by elevated blood sugar levels, which over times can adversely affect the kidney, eyes, heart, blood vessels, heart and nerves. Type 2 diabetes, which frequently affects adults, progress when the body either stops generating enough insulin or develops a resistance to it. The anticipated prevalence of diabetes worldwide in 2019 is 9.3%. (463 million people), however, one in two (50.1%) diabetics is unaware that they have this disease. Both kidney bean and lentil sprouts are rich in phenolic acids and flavonoids like ferulic acid, p-Coumaric acid, sinapinic acid, b-resorcylic acid, chlorogenic acid, catechin, eriodictyol, hesperetin, naringenin, quercetin, kaempferol, isorhamnetin, pelargonidin, cyanidin etc. which have been well recognized for their ant diabetic properties. The modulation of glucose absorption, insulin signaling, insulin secretion, and adipose deposition is supported by the anti-diabetic and anti-obese activity of flavonoids (Vinayagam & Xu, 2015). Flavonoids focus on several molecules that control a number of pathways, including those that enhance cell proliferation, promote insulin secretion, lower apoptosis, and reduce hyperglycemia through regulating liver glucose metabolism (Graf et al., 2005). The suppression of alpha-glucosidase and alpha -amylase, is one of best-known and studied effects of phenolic acid on carbohydrate metabolism (Hanhineva et al., 2010; Manzanaro et al., 2006). Abdel- Moneim et al. (2018) conducted a study on modulation of hyperglycemia and dyslipidemia in experimental type 2 diabetes by gallic acid and p-coumaric acid and reported decrease in serum insulin level in NA/STZ-induced diabetic rats as compared to the control. This comparison was made through homeostasis model assessment (HOMAIR) and quantitative insulin-sensitivity check index (QUICKI). In STZ-induced diabetic rats, GA and PCA can both potentiate the release of insulin from regenerated -cells by blocking the ATP-sensitive K+ channel (Latha & Daisy, 2011). According to Ostadmohammad et al. (Ostadmohammadi et al., 2019) quercetin administration @ 500 mg per day or higher for longer than 8 weeks can significantly lower down the blood sugar levels thus can be an effective recommendation for the management of the diabetes. Lentil sprouts’ effects on cholesterol levels in both obese and overweight diabetic patients was investigated by Aslani et al. (2015) and they concluded that after 8 weeks, the lentil sprout group’s serum HDL-C levels were much higher than those of the comparison group. Additionally, when compared to controls, the lentil sprout group had reduced levels of TG and ox- LDL. These evidences insights the opportunities and further scope of scientifically designed studies to investigate the exact mode of the action of these functional bioactive compounds for the management or treatment of the chronic health disease like diabetes.
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8.2.3 Cardioprotective, Anti-cholesterol and Anti-hypertensive Cardiovascular disease has become a leading cause of disability and premature mortality globally (Roth et al., 2017; WHO, 2021). It was predicted that CVD would be the cause of more than 23 million (about 30.5%) deaths by 2030 worldwide (Lozano et al., 2012). KBS and LS contain good amounts of polyphenols, melatonin, phytosterols, saponins and GABA which are well known for their potential of preventing the cardiovascular diseases in several ways. Besides, reducing oxidative stress, the melatonin supplements may be helpful for reducing inflammation, high blood pressure, and other metabolic syndrome-related signs which has been confirmed through many systematized laboratory findings (Salehi et al., 2019). Phytosterol is yet another component of interest that competes with cholesterol due to their structural similarity and increased lipophilicity. These factors cause phytosterol to limit the micellar solubility of cholesterol in the colon, which significantly lowers enterocyte absorption of cholesterol. Cholesterol transporter proteins thus, help to remove phytosterol from enterocytes into the intestinal lumen by inhibiting ACAT-2 in enterocytes, which prevents cholesterol from being esterified. The transporter proteins viz. Niemann-Pick C1-Like 1 transporter (NPC1L1), transports cholesterol into enterocytes and ATP-binding cassette (ABC) G5/G8 transporters which remove cholesterol from enterocytes and secrete it into bile. Additionally, phytosterols (1–15%) are poorly absorbed while competing with cholesterol for the NPC1L1 transporter. Moreover, the biosynthesis of cholesterol can be decreased by phytosterols via lowering the HMGCS1 gene’s expression. Besides, phytosterols, saponins are also found to be helpful in lowering cholesterol. It is either possible by direct interaction of some saponins with cholesterol leading to insoluble saponin–cholesterol complex or directly by boosting bile acid output from the faeces, helping the small intestine to dissolve cholesterol with more efficacy (Mustafa et al., 2022). By assessing the amount of bile acids in faeces and their prebiotic effect, Micioni di Bonaventura et al. (2017) assessed a hydroalcoholic extract of lentils to look into the hypocholesterolemic effect on an animal study and reported that, cholesterol level of rats was reduced by 16.8%, along with rise of bile acids in their stools, showing that it has the same prebiotic and bifidogenic properties as inulin. Polyphenols on the other hand inhibit angiotensin I-converting enzyme that is responsible for lowering down the blood pressure, showing that hypertension individuals might benefit from using lentil seeds as therapeutic treatments (Hanson et al., 2014). γ-Aminobutyric acid (GABA) is a non-protein amino acid widely existing in both plants and animals. It is known to control blood pressure levels and heart rate, reduce pain and anxiety, and boost the pancreas’ production of insulin, making it a key depressive neurotransmitter. Though, there are limited findings available to claim or justify the application of the lentil and sprouted kidney bean seeds against the cardiovascular diseases yet the presence of the bioactive compounds which has been studied earlier for their cardioprotective, anti-cholesterol and anti-hypertensive justifies their role and symbolic activity against all these diseases. This further represents the new areas of the research which can be explored to clinically document the potential functional properties of the seeds.
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8.2.4 Anti-cancerous The bioactive phytochemicals in legumes have anti cancerous activity which has been reported and documented by AICR (American Institute for Cancer). Lentils and kidney beans have bioactive substances that possess chemoprototective properties possibly by mechanisms of absorption, initiation and destruction of carcinogens, binding them to DNA and fidelity of DNA repair. Moreover, an epidemiological report based on 90,630 women suggests that the ingestion of lentils with high amount of polyphenols can significantly reduce the risk associated with breast cancer (Ganesan & Xu, 2017). Additionally, the use of azoxymethane, reduces of the occurrence of different types of tumours and dysplastic lesions in rat colons (Mustafa et al., 2022). Earlier, saponins were considered as anti-nutritional factors, however, recent systematic findings have projected them as compounds with immense functional properties. Faris et al. (2013) found that saponins are effective as anticarcinogenic agents as they bind with unbound or membrane-bound sterols in the colon and facilitate faecal clearance. Phytosterols, also have anti-cancerous properties against various types of cancers including breast, prostate, colon and other cancers besides their hypocholesterolemic and anti-inflammatory effects. The growth of HT-29 cells’ (human colon cancer tumour cell line) was inhibited after five days of β-sitosterol supplementation at @ 16 mol/L on a human tumour cell line. β-sitosterol modify cell membranes of tumorous cells affecting their integrity and fluidity. Melatonin, a strong antioxidant possess anticarcinogenic and antitumor effects by stimulating systematic cell death, preventing proliferation of cancerous cells by inhibiting different pathways (Jak/Stat, WNT/β-catenin, MAPK, and the Hedgehog pathways), anti-growth signaling and by promoting angiogenesis (Goncalves et al., 2016). These findings suggest strong evidences to project the lentils and kidney beans as potential functional food ingredients having potent anti cancerous properties.
8.3 Methods & Technologies of Sprouting The functional and nutritious components found in grains and pulses can be enhanced using a variety of methods. Sprouting, one of the oldest yet, useful technologies, for enhancing the flavour while it can add a number of additional nutritious elements to the grain. The process of sprouting usually begin with re-hydration of seeds by soaking them in the water under specified treatment condition such as time, temperature, seed weight and water volume ratio. The soaking parameters of seeds depend upon the inherent characteristics of seeds such as seed coat thickness, water absorbing capacity and seed size. After soaking, re-hydrated seeds are incubated for the germination (Gan et al., 2017). Soaking is usually carried out under dark conditions at temperature ranging from 20 to 30 °C for 6 to 24 h and 1:1.5 to 1:20 seed weight to volume ratio (Aguilera, 2014). During germination, water
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applied is sprayed every day on the seeds for maintaining optimum humidity required for seed growth and is replaced twice a day to restrict the microbial growth (Kandil et al., 2015). Water is compulsory to initiate germination and other important biochemical reactions to make the seeds more nutritious an bio functional in nature. Water infiltration also improves seed rehydration, which in turn stimulates the manufacture of gibberellins (GA) in the embryo, which triggers the expression of the hydrolytic enzyme gene (Nelson et al., 2013). The visual indication of the completion of the germination process is the appearance of radicals around the grain embryo, known as sprouts (Bewley & Black, 1994). The germination process beyond activation of GA also assists in the activation of the key enzyme systems including proteases, lipases and various amylolytic enzymes which facilitates the movement of the starches from the amyloplast and release of the secondary metabolites from the vacuole due to changes in the permeability behaviors of the primary cell wall thus ultimately leads to improved nutritional value of the grains.
8.3.1 Pretreatments to Improve Germination There are several ways to speed up the germination process depending on the region being targeted. One of them is priming, which involves the controlled hydration of seeds in order to permit pre-germinative metabolic activity. This technique selection is mostly influenced by plant type, seed form, and physiology of the seed. Osmopriming involves soaking seeds in osmotic solutions such as PEG or salt solutions. Hydropriming involves soaking seeds in predetermined amounts of distilled water or limiting imbibition periods. Hormone priming involves treating seeds with plant growth regulators. Presence of auxin such as IAA in seed sprouting conditions increases germination percentage and coleoptile elongation (Rekoslavskayal et al., 1999; Chiwocha et al., 2005). Thermopriming involves physically treating seeds by pre-sowing them at various temperatures to increase the vigour matrix while, matrix priming involve mixing seeds with water and organic or inorganic solid material in pre-determined proportions. In certain cases, chemical or biological agents are also used to initiate or to enhance the sprouting (Jisha et al., 2013; Paparella et al., 2015). Other cutting-edge technologies, including high pressure processing, magnetic field, ultrasound, ozone processing, pulsed electric field, ultraviolet, non-thermal plasma, microwave radiation, electrolyzed oxidizing water, and plasma activated water, have an impact on the germination and growth traits of seeds with minimum impact on their inherent quality attributes. Application of pulsed electric field as pretreatments promotes opening of plasma membrane cavities, which increases the number of polar molecules in the seed migrating both inward and outward. An enormous number of electric dipoles form inside the seed and align them in the presence of an electric field. Dipole-dipole interaction induced by applied electric field aids in faster respiration, higher water absorption, and improved photosynthetic level, increasing the biological capacity of the seed. As a pretreatment, ultrasonication makes countless tiny holes on the covering and fissures on the pericarp, which
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significantly increases the moisture content of seedlings. The higher holding capacity and high porosity, increases the oxygen availability, which might be the reason of the dominance of sonication as an important and preferable pre treatment before sprouting. A study related to ultrasonic treatment of lentil seeds (Aladjadjiyan, 2011) reported increased germination rate from 92% to 98% after ultrasonication of the lentil seeds @ 42 kHz and 100 W. Ozone application in low concentration also has a positive effect on the timing of seed germination because it breaks dormancy earlier than expected, which often related with the lower levels of abscisic acid in ozone-exposed seeds (Rifna et al., 2019). Plasma activated water is also known to have a positive effect on seed germination due to the production of nitrogen and hydrogen peroxide radicals during water activation. In the physiological processes, hydrogen peroxide not only relieves biotic seed stress but also controls GA and ABA (Zhang et al., 2017a, b). High hydrostatic pressures (HHP, 621 MPa), used as a pre-treatment for soaking lentil, followed by subsequent cooking and has a detrimental effect on the total oligosaccharides content because sugars break down at such high pressures (Han & Baik, 2006). Such novel technologies can pave the future sprouting programs with aim of minimal processing and keeping in mind the delivery of the highest possible and intact quality the masses in general of sufferers in particular.
8.3.2 Effect of Soaking Time and temperature The effects on nutrient content of kidney bean (Phaseolus vulgaris) due to variable soaking time and temperature were investigated in various studies. Nakitto et al., 2015 reported that weight of the kidney bean samples increased with increase in soaking time (from 6 to 12 h). Five hundred grams of the whole seeds were soaked for 12, 24, 36, 48, 60, and 72 h at room temperature in distilled water (ratio 1:30). In general, soaking time increased the content of Na (0.153–0.178 mg/100 g), Ca (0.2–0.237 mg/100 g), K (0.467–0.643 mg/100 g), P (0.251–0.345 mg/100 g), and Mg (0.221–0.254 mg/100 g) compared to the values obtained in the untreated beans. However, soaking for 60 h produced the best results for vitamins A and C (581.35 and 32.15 mg/100 g) respectively, while vitamin B (3.16 mg/100 g) produced best outcomes at 72 h (Alu & Ahiwe, 2018). The nutritious and anti-nutritional components of lentil flours are significantly affected by short-term soaking with variations in the temperature, soaking medium, lentil-water ratio, milling of the seeds and illumination (Vidal-Valverde et al., 2002). Protein efficiency ratio (PER) of sprouted lentil seeds sprouted for 6 days was 2.94 which was about 1.08 times as high as that in control (Fouad & Rehab, 2015). Whole green lentils which were soaked for 16 h has 5 times more total phenolics and 3 times more saponins, as well as a sizable amount of hydrolysis products such galactose, glucose, and fructose (Huang et al., 2018). This shows that the endogenous alpha-galactosidase lost in the lentil seeds’ soaking solution can have a prebiotic effect that would benefit human health and well-being (Njoumi et al., 2019). Hojjat and Galstayan (2012) reported that with an
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increment in temperature values the rate of sprouting also increases to some extent and then starts decreasing. They conducted a study in which lentil seeds were subjected to different temperatures (39, 46, 54, 61, 68 and 75 °F) and maximum germination was obtained 88% at 68 °F, concluding that temperature is an exceedingly important factor in seed sprouting. Above mentioned studies are proof that variable soaking time and temperature is very cost-effective treatment to raw grains as it play pivot role in enhancing the various functional and nutritional properties of sprouts.
8.3.3 Effect of Other Elicitors on Composition of Sprouts A chemical known as an elicitor causes the plant to experience hypersensitivity. They can be derived from both abiotic and biotic sources (UV radiations, heavy metals, bacteria, fungi, thermal stress). Swieca et al. (2016) investigated the oxidative elicitation using hydrogen peroxide (20 mM and 200 mM solutions) as an abiotic elicitor. When the stressful condition was introduced two days after sowing, the highest levels of phenolics in seedling tissues, especially in terms of chlorogenic, ferulic, o-coumaric, and salicylic acids was observed. It also increased the ability to protect lipids against peroxidation by between a factor of 12 and 8 respectively. The addition of sodium selenite solutions (5–10 ppm) to textile germination beds for wheat seeds resulted in wheat seedlings with higher levels of vitamin C and antioxidant activity (Moldovan et al., 2011). During germination, the utilisation of oligosaccharides and polysaccharides has attracted the greatest attention among biotic elicitors. Many non-graminous species have been studied to see how chitosan affects bean (Mendoza-Sánchez et al., 2016), soybean (Khan et al., 2003), lentil (Peñas et al., 2015) broccoli (Barrientos Carvacho et al., 2014) and lettuce (Viacava et al., 2015). Utilization of elicitors for sprouting is still an evolving area, however, there are limited studies that objectify the systematic role of elicitors, and still strong evidences are required to understand their mode of action and mechanism and utilization.
8.4 Novel Methods for the Processing of Sprouts Nowadays, people look for minimally processed foods to have least losses to the functional food compounds and have the high nutrition packed meals. So far, processing from the perspective of nutrition and functionality has a bad image in the consumers mind, but as far as pulses are concerned, processing helps to get rid of anti-nutritional factors from them thus, can be a crucial step for making them healthy foods. Among various processing methods the novel methods i.e. high pressure processing, ultrasound, irradiation etc. have been exploited recently by researchers around the globe as they maintain quality of the treated product and have minimum losses to the bioactive compounds present in the grains. Foods
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processed under high pressure undergo a number of modifications, such as protein structure and coagulation, altered starch melting characteristics, rearrangement of polymorphic lipid forms, inactivation of microbes, and induction of low temperature chemical modification. It causes pulses to have more water solubility with enzyme activity (endogenous), which significantly reduces their anti-nutritional (phytic acid and tannin) content (Alsalman & Ramaswamy, 2020). At high-pressure (500 MPa for 10 min) germinated lentil seeds show reduced contents of the non- digestible oligosaccharides. After 14 days of storage at 4–8 °C, the -galactosides decreased from 16% to 5% and then further to negligible amounts (Dostalov et al., 2009). Additionally, Hall and Moraru (2021) conducted a comparative research on the impact of thermal treatment and high-pressure processing on protein quality, trypsin inhibitor activity and protein digestibility and reported that treatment at 95 °C for 15 min dramatically reduced the protein digestibility while treatment at 600 MPa for 4 min at 5 °C resulted in increased stomach proteolysis. Besides, high- pressure processing leads to minor reduction in trypsin inhibitor activity (8%), but heat treatment resulted in a significantly greater reduction (86%), with no discernible difference in the total protein quality or digestibility (in vitro) for either. Also, high-pressure processing has shown an increase in ACE inhibitory activity that is beneficial to health (Dostalov et al., 2009). Additionally, combination of commercial protease (Corolase 7089) and high pressure has been utilized for enzymatic hydrolysis of lentil flours that led to a maximum inhibition of ACE (70.77%) at 300 MPa (Garcia-Mora et al., 2015). Therefore, the high pressure improves the accessibility of enzyme towards substrate and expose to new residue targets which release bioactive sequences. Radiation is yet another method of processing that can improve the nutritional value of legumes, along with significant reduction or elimination of anti-nutrients by triggering the breakdown of sugars without affecting the material’s functionality or palatability. El-Niely (2007) assessed the effect of radiation processing on several anti-nutrients in lentils. The treatment of lentils @ 5, 7.5, and 10 kGy considerably reduced the phytic acid content by 15.1%, 25.2% and 32.7% and the tannin content by 7.6%, 12%, and 21.7% respectively. The tannin content and phytic acid was reduced with linearly increased radiation dose. Aylangan and Ozyardimci (2017) reported that raffinose and stachyose were severely damaged by other anti-nutrients at various irradiation dosages (0.25, 0.50, and 1.0 kGy), with no reported effect after storage for 6 and 12 months. The presence of oligosaccharides indirectly affects another aspect of how irradiation affects during the storage-induced retrogradation in lentil starch. Cavitation caused by ultrasonic processing results in both unstable and stable bubbles inside the edible product. Energy builds up in hot places during the collapse of these cavitation bubbles, creating extreme circumstances that cause both shear energy waves in cavitation zone (Karaman et al., 2017). As a result, the conditions in ultrasonic processing cause a variety of alterations in the food item. However, research indicates that these circumstances are insufficient to impart significant
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impact on phytic acid and resistant starch (Kaya et al., 2017). Also, due to enhanced oligosaccharide leaching during the procedure, ultrasonic soaking is a successful reduction approach for other anti-nutrients like oligosaccharides (Han & Baik, 2006). According to a study by Han and Baik (2006), soaking uncooked lentils in ultrasonic water at 47 MHz for 90 min reduced oligosaccharides by 36.36–57.76% raffinose, 53.80–74.09% ciceritol and 50.35–53.66% stachyose. Another systematic study, demonstrated that anti-nutrients such saponins and sapogenins were recovered from lentil and employed as bioactive compounds using a probe type ultrasound with 1/2″ diameter for extraction with sonication output 60% amplitude and 75 °C temperature (Del Hierro et al., 2018). Usage of these novel techniques help in decreasing the anti-nutritional content present in pulses without affecting their nutrition profile. Keeping in mind the losses of compounds due to conventional processing, more efficient researches are required, so that lab technologies can be used in commercial world.
8.4.1 Storage and Packaging During the storage of food at low temperature, the rates of senescence as well as growth of microbial deterioration decreases which helps to prevent quality loss (Mir et al., 2017; Raimondi et al., 2017). Berba and Uchanski (2012) studied the metabolic activity and shelf life of radish, arugula and red cabbage seedlings (without radicles) and reported that, radish had a longer storability than the other species, regardless of whether kept at 10 °C (14 vs. 7 days on average) or at 4 °C (21 vs. 14 days on average). The influence of storage temperatures must be assessed in conjunction with other factors that affect shelf life which includes washing of sprouts prior to packaging and atmospheric composition during packaging (hurdle technology). For instance, by lowering O2 and raising CO2 partial pressures in the package headspace, variable atmosphere packaging significantly extends the seedlings’ shelf life (Vale et al., 2015). The oxygen transmission rate (OTR) of packaging film, respiration rate of product, package surface area, product weight and storage temperatures are some of the variables that affect the package atmosphere (Sandhya et al., 2010). However, package size and product weight are typically predefined in food supply chains, choosing a packaging film with a suitable OTR could be an appropriate method to maintain quality and extend shelf life (Xiao et al., 2014). A study conducted by Kou et al. (2013) concludes that when buckwheat sprouts (without radicles) were preserved at 5 °C in 16.6 pmol/s/m2/Pa ORT films, they performed their best for 21 days while preserving the highest quality characteristics and tissue integrity.
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8.4.2 Value Addition Modern day consumers are especially concerned about foods that have a long shelf life, excellent sensory qualities, and high bioavailability. Numerous studies have emphasized the health benefits of biologically stimulated or sprouted grains, and if these grains can be employed in traditional diets, they will boost immunity, balance the relationship between acid and base, and promote metabolism. Because of the loosened texture and enhanced flavors, the development of biomaterial compounds during the biological activity of grains contributes to the improvement of organoleptic properties. Cookies from germinated triticale, kidney beans, and chickpeas were prepared by Sibian and Riar (2020) comprising 15.13 g of germinated kidney bean and 34.50 g of chickpea flour per 100 g cookies flour. Composite flour cookies demonstrated enhanced protein and carbohydrate digestion. Due to the substitution of legumes and germination, the essential amino acid content of the optimized cookies was higher than that of the control (wheat flour). In a recent study, Bhavya and Prakash (2021) evaluated nutritional properties of iron fortified flatbreads enriched with sprouted greens and legumes (chickpea, green gram). It was concluded that flatbread enhanced with greens and iron salts had a greater protein content than the control (12.55 g/100 g), ranging from 12.82 to 13.42 g/100 g dwb. Additionally, in comparison to controls, flatbreads made from legumes also had higher concentrations of calcium (88.69–94.83 mg/100 g) and iron (14.77–16.98 mg/100 g).
8.5 Conclusion Lentils and kidney beans are considered as nutritious and health-promoting foods. Sprouting is the most economical method of processing the seeds to improve their nutritional and functional value by synthesis of essential nutrients as well as reducing the anti-nutritional factors. Further, it enhances the bioactivity of the lentils and kidney beans by production of novel bioactive compounds such as melatonin and γ-aminobutyric acid (GABA), also enhancing the quantities of existing phytochemicals viz. phenolic compounds, phytosterols and glucosinolates. Most of these compounds have antioxidant properties and exhibit many health promoting properties including anti-diabetic, immunomodulatory, anti-aging, cardio protection etc. Especially lentils and kidney beans sprouts are rich sources of melatonin and γ-aminobutyric acid (GABA), however, currently many studies are focusing on these compounds. The advancement in sprouting technology (pretreatment with harmones, salt solutions, high-pressure processing, pulsed electric field, ultrasound, ozone processing, ultraviolet, magnetic field, microwave radiation, non-thermal plasma, electrolyzed oxidizing water, plasma activated water and other biotic and abiotic elicitors) also promotes the quality and hygienic production of lentils and kidney beans sprouts as well as novel processing and packaging methods are helpful enough for their preservation and value addition. Therefore, by considering the fact
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that sprouts are gaining popularity recent days, lentils and kidney beans sprouts have potential to meet the expectations of the consumers regarding availability of various health promoting compounds in the foods they eat. The major challenges to scale up the production of sprouts are the chances of contamination and related outbreaks which needs to be investigated in future studies. Systematic studies involving animal-based models for defining the bioavailability and bioactivity of specific target phytochemical needs clinical validation and scientific documentation to further increase the consumption and provide cheaper and healthy food options to the society.
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Chapter 9
Clover and Alfalfa Sprouts Bababode Adesegun Kehinde, Oluwakemi Igiehon, Adekanye Oluwabori, and Ishrat Majid
9.1 Alfalfa Sprouts: Basic Introduction Medicago sativa, commonly referred to as alfalfa, lucerne, blalusern or even lyutzernaposevnaya in different parts of the world, is a leguminous angiosperm plant belonging to the Fabaceae family. It is perennial and is thought to originate from south-central Asia though it is available on the majority of continents around the world. It is cultivated in diverse countries such as Russia, Morocco, Lebanon, Libya, United States, Peru, Kora, Estonia, Italy, Albania, and Germany, amongst several other countries. As a forage legume, alfalfa can live up to eight years on average, though longer life spans have been reported as being associated with the climatic conditions and genetic factors. It grows at a gradual but steady rate from small-sized seeds (1–2 mm) with protruding roots that contain shoot buds for its regrowth in the event of harvesting or grazing. Alfalfa is versatile in its dietary usage as it is commonly used beyond human consumption as silage or hay for animal feeding. Dairy animals, horses, sheep and other cattle as well as poultry and rabbits are fed with processed alfalfa derivatives based on its considerable amounts of carotenoids, protein and fiber. Humans consume alfalfa in different processed forms such as sprouts B. A. Kehinde (*) Food Processing Center, University of Nebraska-Lincoln, Lincoln, NE, USA O. Igiehon Department of Microbiology and Immunology, Louisiana State University, Shreveport, LA, USA A. Oluwabori Department of Veterinary and Biomedical Sciences, Mississippi State University, Starkville, MS, USA I. Majid Department of Food Technology, Islamic University of Science and Technology, Awantipora, JK, India © The Author(s), under exclusive license to Springer Nature Switzerland AG 2023 I. Majid et al. (eds.), Advances in Plant Sprouts, https://doi.org/10.1007/978-3-031-40916-5_9
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and dehydrated powders. Its sprouts are edibles as sandwiches and salads and its powders are used for tea mixes.
9.2 Alfalfa Sprouts: Phytochemistry, Biological and General Usage The phytochemistry of alfalfa sprouts are strongly affected by their germination conditions and responsible for their biological functionalities on consumption. Factors such as germination time, lighting conditions, temperature, and relative humidity of the environment have been studied to affect the use of alfalfa sprouts. Their processing procedures and parts examined could also affect their compositional findings (Fig. 9.1). Numerous biofunctional phytochemicals have been identified from alfalfa sprouts such as minerals (selenium, copper, manganese), vitamins (ascorbic acid, beta-carotene, and tocopherol), essential and non-essential amino acids (threonine, lysine, glutamic acid, serine, phenylalanine, histidine, and threonine), steroids (loliolide, isoliquiritigenin, liquiritigenin, and coumestrol), saponins (soyasapogenols, azukisaponin, 3–0-glucoside, and medicagenic acid) and several other volatile components (trans-2-pentenal, trans-3 hexenylbutanoate, trans-3-hexenylacetate, and methyl phenyl ketone) (Hong et al., 2011; Al-Snafi et al., 2021). Alfalfa sprouts have been studied for their various food and health benefits and functionalities. Mattioli et al. (2016) sprouted alfalfa seeds on moistened tissue paper placed in a dark environment with temperature controlled at 20 °C for three days. Fresh sprouts obtained were then supplemented to a standard chicken feed
Whole Meal
Leaf Meal
Moisture 12%
Ash 14%
Ash 19% Protein 26% Fibre 29%
Fibre 44% Fat 4%
Moisture 14%
Protein 34%
Fat 4%
Fig. 9.1 Comparison of proximate composition between whole and leaf meals of alfalfa sprouts. (Data obtained from Mielmann, 2013)
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diet at a rate of 40 g/d and the eggs laid by the chicken examined. Results obtained reported a reduction in cholesterol and plasma of the eggs; a trend attributed to a probable synergy between different compounds of the sprouts such as sterols, isoflavones, lignans and polyunsaturated fatty acids. In addition, such eggs were found to possess higher amounts of phytoestrogens (isolariciresinol, equol, and daidzein), carotenes (zeaxanthin, lutein, and β-carotene), vitamins (retinol, α-, γ-tocotrienol, and α-tocopherol) and omega-3 polyunsaturated fatty acids relative to the control having the standard diet only. Almuhayawi et al. (2020) sprouted alfalfa seeds with photosynthetically active radiation (PAR) of 400 μmol m − 2 s − 1 with white, fluorescent tubes at a 16/8-day night photoperiod. However, the germination occurred at elevated carbon dioxide levels of 620 ± 42 μmol CO2 mol − 1air ppm and ambient carbon dioxide levels of 400 ± 27 μmol CO2 mol − 1 air while keeping the temperature and relative humidity at steady values of 25 °C and 60% respectively. The carbon dioxide elicitation was found to improve the process of photosynthesis and the pigment concentrations of the sprouts along with levels of flavonoids, vitamins, minerals, and phenolics. Phenolics and flavonoids such as narigenin, apigenin, myricetin, vitexin, daidzein, rutin, luteolin, quercetin, kampferol, catechin, caffeic acid, coumaric acid, ferulic aicd, and galic acid were majorly higher for elevated carbon dioxide samples. Total proteins, free amino acids, L-canavanine, carbohydrates, total lipids, fibres, and mineral profile (sodium, phosphorus, potassium, magnesium, manganese, zinc, iron, copper, and calcium) were also majorly enhanced by the elevation of carbon dioxide levels. Plaza, de Ancos and Cano (2003) developed a drying procedure termed as controlled instantaneous pressure release or “détente instantanée contrôlée (DIC)”. The alfalfa seeds were heated under 2–10 bar at a temperature of 200 °C for less than 60 s after which the pressure was immediately released. Thereafter, the seeds were soaked normally and sprouted for a 96-h period at 28 °C in the dark. Contents of phytoestrogens (genistein and daidzein), vitamins such as ascorbic acid, pyridoxal, riboflavin, thiamine, and α-tocopherol, and minerals such as zinc, sodium, manganese, magnesium, potassium, iron, iron, copper and calcium were reportedly increased for the sprouted seeds that were dried by the procedure. Aloo, Ofosu, and Oh (2021) conducted an extensive phytochemical examination of alfalfa sprouts germinated at temperatures of 28–30 °C in the dark for 5 days. The sprouts were then oven-dried pulverized into small-sized fine powder particles and analyzed using the Ultra-High-Performance Liquid Chromatography-Triple/Time-Of-Flight Mass Spectrometry (UHPLC-Q- TOF-MS/MS) technique. Numerous metabolites were identified such as catechin gallate, actinonin, epicatechin, L-tryptophan, L-arginine, fumaric acid, n-methylglutamic acid, pyroglutamic acid, adenosine, pyroglutamic acid, 3-furoic acid, uridine, malic acid, 1-hexadecylamine, galactaric acid, gluconic acid, dl-o- tyrosine, trans-cinnamic acid, L-tryptophan, pyroglutamic acid, D-ornithine, γ-aminobutryic acid, and DL-homoserine. Debski, Wiczkowski, and Horbowicz (2021) examined the effect of sodium metasilicate and iron chelate elicitation on the sprouting of alfalfa. High pressure sodium lamps were used for photosynthetically active radiation to provide light conditions 100–120 μmol/ (m2 ·s) at 16 ± 2 °C (night, 8 h) and 20 ± 1 °C (day, 16 h) for seven days. High-Performance Liquid
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Chromatography/Electrospray Ionization Tandem Mass Spectrometry (HPLC/ ESI-MS/MS) was used to analyze phenolic compounds and the study reported that elicitors caused an increase in the content of apigenin, luteolin and quercetin glycosides. In addition, levels of caffeic acid, p-hydroxybenzoic, p-hydroxycinnamic, and ferulic acid esters were also found to increase.
9.3 Clover: Basic Introduction It is commonly referred to as trefoil and since it is an angiosperm belonging to the Fabaceae family, it is taxonomically related with Alfalfa. The clover is a legume with hundreds of species of flowering plants known for its prominent diversity in temperate regions, though it has been found in Africa and Southern America. Unlike alfalfa, clover can be biennial, annual or briefly-perennial., growing to about 11.5–13.0 in plant height and with diverse leaf numeration such as sepatfoliolate, hexafoliolate, quinquefolilolate, trifoliolate, bifoliolate, and monofoliolate, with the quatrefoilate having the rarest occurrence. Red clover (Trifolium pratense) and white clover (Trifolium repens) are the most cultivated, though other clover species such as hare’s-foot trefoil (Trifolium arvense), zigzag or meadow clover (Trifolium medium), strawberry clover (Trefolium grafiferum), swedish clover (Trifolium hybridium), hop trefoil clover (Trifolium campestre), and lesser hop trefoil (Trifolium dubium) also exist. In vegetations, clovers are more productively pollinated by bees because clover blooms are proficient sources of nectar. However, clover plants are affected by soil pH, diseases, and pests and can be short-lived.
9.4 Clover Sprouts: Phytochemistry, Biological and General Usage Beyond their environmental uses as legumes for nitrogen fixation and compost making for soil replenishment, clovers are serviceable as forage and food for humans when processed. However, they have been studied for their phytochemistry and health benefits especially after beensprouted (Gałązka-Czarnecka et al. 2020). Tahany et al. (2021) used Gas Chromatography-Tandem Mass Spectrometry (GCMS/MS) to examine the phytochemicals of sprouted clover seeds. The soaked seeds were sprouted at room temperature for a 72-h period, subsequently oven-dried for 48 h at 55 °C and milled. Several phytochemicals such as tarpenoids and their derivarives, phenols and their derivatives, and flavonoids and their derivatives were reported to have increased due to the sprouting. They include: nobiletin, nerolidol, vitexin, geranly isovalerate, 2′,3′-Dimethoxyflavone, Luteolin 5,7,3′,4′ -tetramethylether, Isolongifolol, Thunbergol, Afromosin 7-O-glucoside, 5,7,3′,4′,5′ -pentahydroxyflavone, levomenthol, and citronellyl tiglate. Metabolites such as docasane,
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4-methyl, 3,7,8,2′-tetramethoxyflavone, and 7,8,3,4-tetramethoxyflavone were identified in the sprouts and not in the seeds. Chiriac et al. (2020a, b) examined the effects of sprouting on the mineral profile of red clover that was germinated for 96 h in the dark at 80% relative humidity and 25 °C. Significant increases were reported for all minerals due to sprouting in comaaprison with unsprouted seeds. Manganese increased from 7.87 ± 0.5 to 8.95 ± 0.8 μg/g DW, nickel increased from 2.50 ± 0.5 to 3.39 ± 0.2 μg/g DW, selenium increased from 6.16 ± 1.4 to 8.76 ± 0.6 μg/g DW, copper increased from 16.04 ± 1.1 to 19.96 ± 3.2 μg/g DW, zinc increased from 39.94 ± 3.3 to 49.50 ± 1.2 μg/g DW, iron increased from 50.61 ± 3.2 to 114.68 ± 3.0 μg/g DW, potassium increased from 8186.36 ± 72 to 16963.41 ± 7.6 μg/g DW, sodium increased from 223.61 ± 4.2 to 2603.66 ± 9.9 μg/g DW, magnesium increased from 2512.96 ± 5.8 to 3347.56 ± 2.7 μg/g DW, and calcium increased from 250.83 ± 6.9 to 601.59 ± 5.6 μg/g DW. The total sum of examined minerals for unsprouted red clover seeds was 11296.88 ± 3.9 μg/g DW and 23721.46 ± 3.5 μg/g DW for sprouted ones. Yokoyama et al. (2020) studied the effects of red clover administration on the in vivo prevention of metabolic syndromes caused by western diet. Red clover seeds were germinated for a 5-day period, lyophilized, and orally administered for 8 weeks to eight-week-old male C57BL/6 J mice fed with high-fat and high- carbohydrate diet. Plasma biochemistry components such as fasting plasma glucose, low density lipoprotein cholesterol, high density lipoprotein cholesterol, total cholesterol, and total glyceride showed significant decreases for the animal study group fed with the western diet and sprouted clover relative to those fed with the western diet alone. Budryn et al. (2018) investigated how sprouting affected the estrogenic potential of red clover. The seeds were sprouted in varying conditions of white light (310 nm at light 24 h/day, or 340 nm, 12 or 24 h/day), and temperature (18 or 25 °C) at a relative humidity 80% for a 10-day period. Liquid Chromatography- Electrospray Ionization-Mass Spectrometry (LC-ESI-MS) was used to identify biological compounds such as equol, daidzein, genistein, biochanin A, formonentin, and β-estradiol. Chirac et al. (2020a, b) in another study examined the presence of biologically active polyphenols in clover sprouts using the UHPLC-Q Exactive Hybrid Quadrupole Orbitrap High-Resolution Mass Spectrometry. Isoflavones such as glycitein, formononetin, ononin, daidzein, and genistein, phenolic acids such as syringic, p-coumaric, abscinic, ellagic, ferulic, caffeic, chlorogenic, and gallic acids along with flavonoids such as hyperoside (quercetin 3-galactoside), galangin, myricetin, chrysin, pinocembrin, pinostrobin, hesperitin, naringin, naringenin, isorhamnetin, kaempferol, apigenin, rutin (quercetin3-rutinoside), quercitin, epicatechin and catechin were identified.
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9.5 Conclusion Alfalfa and clover plants are available in several countries of the world and despite being prominent as forages for animal feed, they are typically exploited for their numerous benefits. In addition to being legumes aid in soil nitrogen fixation, they are naturally loaded with biologically functional phytochemicals of potential health benefits, especially against metabolic syndrome disorders. Recent studies have shown that sprouting their seeds improves the profile of their concomitant nutraceuticals such as isoflavones, phenolics, flavonoids and other phytoestrogens. Sprouting conditions such as time, temperature, relative humidity, seed pre-treatment and presence of one or more elicitors have been reported to be of significant effects on the metabolism of the bioactive compounds derivable due to sprouting. Scientific investigations with finesse chromatographic techniques have also revealed that remarkable changes occur in the presence and content of phytochemicals when alfalfa and clover seeds are sprouted. Assuredly, sprouting is one important technique for consideration when the goal is to improve the food and health serviceability of alfalfa and clover.
References Almuhayawi, M. S., Hassan, A. H. A., Al Jaouni, S. K., Hussien, M., Alkhalifah, D., Hozzein, W. N., Selim, S., Abdelgawad, H., & Khamis, G. (2020). Influence of elevated CO2 on nutritive value and health-promoting prospective of three genotypes of Alfalfa sprouts (Medicago Sativa). Food Chemistry, 340, 128147. https://doi.org/10.1016/j.foodchem.2020.12814 Aloo, S.-O., Ofosu, F.-K., & Oh, D.-H. (2021). Effect of germination on alfalfa and buckwheat: phytochemical profiling by UHPLC-ESI-QTOF-MS/MS, bioactive compounds, and in-vitro studies of their diabetes and obesity-related functions. Antioxidants, 10, 1613. https://doi. org/10.3390/antiox10101613 Al-Snafi, A., Hanaa, S., Khadem, Hussein, H. A., Alqahtani, A., Batiha, G., & Jafari Sales, A. (2021). A review on Medicago sativa: A potential medicinal plant. International Journal of Pharmaceutical and Biological Science Archive, 1, 22–033. https://doi.org/10.30574/ ijbpsa.2021.1.2.0302 Aly, T., Mustapha, A., Zhang, L., Yu, X., Yagoub, A. E. G., Ma, H., Chen, L., & Zhou, C. (2021). Interaction effects of salinity and ultrasound pretreatment on the phytochemical compounds of clover sprouts (Vol. 5, pp. 90–101). https://doi.org/10.31080/ASNH.2020.05.0838 Budryn, G., Gałązka-Czarnecka, I., Brzozowska, E., Grzelczyk, J., Mostowski, R., Żyżelewicz, D., Cerón-Carrasco, J. P., & Pérez-Sánchez, H. (2018). Evaluation of estrogenic activity of red clover (Trifolium pratense L.) sprouts cultivated under different conditions by content of isoflavones, calorimetric study and molecular modelling. Food Chemistry, 245, 324–336. https://doi. org/10.1016/j.foodchem.2017.10.100 Chiriac, E. R., Chiţescu, C. L., Borda, D., Lupoae, M., Gird, C. E., Geană, E.-I., Blaga, G.-V., & Boscencu, R. (2020a). Comparison of the polyphenolic profile of Medicago sativa L. and Trifolium pratense L. Sprouts in different germination stages using the UHPLC-Q Exactive hybrid quadrupole Orbitrap high-resolution mass spectrometry. Molecules, 25, 2321. https:// doi.org/10.3390/molecules25102321 Chiriac, E. R., Chiţescu, C. L., Sandru, C., Geană, E.-I., Lupoae, M., Dobre, M., Borda, D., Gird, C. E., & Boscencu, R. (2020b). Comparative study of the bioactive properties and elemental
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composition of red clover (Trifolium pratense) and Alfalfa (Medicago sativa) sprouts during germination. Applied Sciences, 10, 7249. https://doi.org/10.3390/app10207249 Debski, H., Wiczkowski, W., & Horbowicz, M. (2021). Effect of elicitation with iron chelate and sodium metasilicate on phenolic compounds in legume sprouts. Molecules, 26, 1345. https:// doi.org/10.3390/molecules26051345 Gałązka-Czarnecka, I., Korzeniewska, E., Czarnecki, A., Kiełbasa, P., & Dróżdż, T. (2020). Modelling of carotenoids content in red clover sprouts using light of different wavelength and pulsed electric field. Applied Sciences, 10(12), 4143. https://doi.org/10.3390/app10124143 Hong, Y.-H., Wang, S., Hsu, C., Lin, B.-F., Kuo, Y.-H., & Huang, C. (2011). Phytoestrogenic compounds in alfalfa sprout (Medicago sativa) beyond Coumestrol. Journal of Agricultural and Food Chemistry, 59(1), 131–137. https://doi.org/10.1021/jf102997p Mattioli, S., Dal Bosco, A., Martino, M., Ruggeri, S., Marconi, O., Sileoni, V., et al. (2016). Alfalfa and flax sprouts supplementation enriches the content of bioactive compounds and lowers the cholesterol in hen egg. Journal of Functional Foods, 22, 454–462. https://doi.org/10.1016/j. jff.2016.02.007 Mielmann, A. (2013). The utilisation of lucerne (Medicago sativa): A review. British Food Journal, 115(4), 590–600. https://doi.org/10.1108/00070701311317865 Plaza, L., de Ancos, B., & Cano, P. M. (2003). Nutritional and health-related compounds in sprouts and seeds of soybean (Glycine max), wheat (Triticum aestivum. L) and alfalfa (Medicago sativa) treated by a new drying method. European Food Research and Technology, 216(2), 138–144. https://doi.org/10.1007/s00217-002-0640-9 Yokoyama, S.-I., Kodera, M., Hirai, A., Nakada, M., Ueno, Y., & Osawa, T. (2020). Red clover (Trifolium pratense L.) sprout prevents metabolic syndrome. Journal of Nutritional Science and Vitaminology, 66, 48–53. https://doi.org/10.3177/jnsv.66.48
Chapter 10
Black-Eyed Peas, Chickpeas and Pea Sprouts Meenakshi Trilokia, Wani Suhana Ayoub, and Preeti Choudhary
10.1 Introduction Sprouting (also known as sprouts) is the process of soaking grains in water and allowing them to rest in moist conditions until they begin to germinate. Sprouting has been shown to improve nutritional quality by increasing protein and starch digestibility, vitamin content, and mineral bioavailability, while decreasing anti- nutritional components including phytic acid, tannins, and galacto-oligosaccharides (stachyose and raffinose) (El-Adawy, 2002; Mahadevamma & Tharanathan, 2004). According to the American Association of Cereal Chemists (AACC) states that grains that have been germinated are to be regarded as complete grains if they contain all of the bran (fiber), germ (embryo), and endosperm (albumin), the sprout length does not surpass the length of the grain, and the nutritional benefits not compromised. The prerequisite for the germination process is the soaking of grains. Because seeds absorb water during soaking, the latent condition of the seeds is disrupted (Bewley, 1997). There are three steps to the germination process for seeds: firstly, the imbibition stage, in which the seeds rapidly absorb water; secondly, the initiation of biochemical process; the reduction in water absorption rate; and lastly, in which the radicle emerges from seed (Wolny et al., 2018). Meanwhile, the sprouting restarts the seed metabolism, this will results in the catabolism and degradation of macronutrients and antinutritional components, also the biosynthesis of secondary metabolites with possible health benefits (Penas E & M. Trilokia (*) Division of Food Science and Technology, SKUAST-Jammu, Jammu, Jammu and Kashmir, India W. S. Ayoub Desh Bhagat University, Mandi Gobindgarh, Punjab, India P. Choudhary KVK, Reasi, SKUAST-Jammu, Jammu, Jammu and Kashmir, India © The Author(s), under exclusive license to Springer Nature Switzerland AG 2023 I. Majid et al. (eds.), Advances in Plant Sprouts, https://doi.org/10.1007/978-3-031-40916-5_10
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Martnez-Villaluenga C, 2020). This causes a number of changes to take place during the germination process. As a result, germination is a quick and low-cost procedure that increases the flavour, digestibility, and availability of several nutrients (Bains et al., 2014).
10.2 Improvement in Nutritional Value Sprouts are a fantastic source of numerous macronutrients, micronutrients, secondary plant metabolites, primarily glucosinolates and phenolic substances (Yang et al., 2013). The biological activity of these substances is associated with their antioxidant capacity that can lessen the oxidative stress brought on by the abnormally higher level of reactive oxygen species within the cells. This capacity also provides molecular tools to balance the unevenness between the creation of reactive oxygen species and the capacity to curb the redox balance (Abellan et al., 2019). This composition enables sprouts to deliver advantageous bioactive components once they are regularly included in the diet (Abellan et al., 2019). Thus, the germination process is a useful way to organically increase the nutritive value of grains, and it is crucial in nations wherever diet-related disorders are widespread (Nelson et al., 2013). During the germination process, the grains stimulate and produce endogenous enzymes that consequently encourage the escape of the kernel pods following the water absorption and time of respiration (Ma et al., 2020). A dynamic and complicated movement of nutrients including remobilization, breakdown and accumulation is reported to be accompanied by substantial phytochemical changes as a result of the action of the endogenous enzymes (Nelson et al., 2013; Makinen & Arendt, 2015; You et al., 2016). The phenolic compounds originate from the polyketide pathway(s) and/or the phenylpropanoid-derived shikimate, both of which have many phenolic rings and don’t have nitrogen based functional groups in their most fundamental structures (Abellan et al., 2019). Cells are shielded from oxidative damage by phenols (Podsedek, 2007). Their capacity to remain stable in various conditions, as well as the quantity and distribution of hydroxyl groups, all influence their antioxidant effectiveness (Podsedek, 2007). Secondary metabolites known as phenolic acids are frequently present in meals obtained from plants (Roche et al., 2017). The most prevalent and varied class of polyphenols are flavonoids (Roche et al., 2017). Foods high in phenolic component can lower the threat of a variety of health related issues (Singh et al., 2017). According to Singh et al. (2017), these chemicals not only add to the organoleptic qualities of food furthermore offer a number of health advantages. Generally speaking, grain germination increases overall polyphenol content; however, this varies by species and sprouting circumstances (Benincasa et al., 2019; Paucar-Menacho et al., 2017). Vitamins, g-aminobutyric acid (GABA), and polyphenols are just a few of the bioactive chemicals that can accumulate during the germination process in seeds
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and sprouts (Gan et al., 2017). During the germination phase, the bioactive substances may be created from scratch or changed in some other way (Gan et al., 2017). It should be emphasized that the majority of research assess the nutritive value of freshly grain sprouts because this is the primary eating method. Additionally, it’s crucial to understand how the nutritional makeup of sprouts varies when they go through certain culinary operations, including cooking. When compared to raw seeds, germination process increases the amount of some water-soluble vitamins namely, folate in soybean seeds and green bean seeds, vitamins B1 and B6, and vitamin C in sprouted buckwheat, chickpeas, lupins, mung beans, and soyabeans as well as their sprouts. Germination also increases the content of phenolic compounds in various seeds (Gan et al., 2016, 2017). When it comes to the vitamins (fat-soluble) germination can change the amount of vitamin E in the edible seeds like soybeans (sprouted), but this impact was not seen in other seeds (Gan et al., 2017). Since storage proteins are degraded into peptides and amino acids by proteolytic enzymes after 2–3 days of imbibition, a shift in protein composition is another significant alteration brought on by grain germination (Benincasa et al., 2019). Higher concentrations of important amino acids which are involved in the amalgamation of proteins in the human were also released (Benincasa et al., 2019). In terms of carbohydrates, sprouting increases the amylase activity, which is in charge of hydrolyzing starch into simple sugars; moreover, the length of the germination period influences the starch degradation (Marti et al., 2021). A unique maize cultivar’s alterations in starch structure and in vitro digestion profile between 0 and 5 days after germination were examined by (Ma et al., 2020). As maize germination continued, the data revealed a considerable decrease in the overall concentration of starch, amylose, and amylopectin (Ma et al., 2020). Triacylglycerols, an oil-like lipid, are widely distributed throughout the live tissues of whole grains found in cereals, such as the embryo, scutellum, and aleurone (Benincasa et al., 2019). During the germination of seeds, lipids are converted into energy (Xu et al., 2020a, b). In a nutshell, the activation of lipase caused the triacylglycerols to begin hydrolyzing into the free-fatty acids moreover, increasing the ratio of saturated/unsaturated fatty acid (Xu et al., 2020a, b; Benincasa et al., 2019). Numerous studies have shown that the sprouting process has positive impact on the nutritive composition of seeds. By increasing the vitamin C and E concentration, protein digestibility, antioxidant capacity, and reducing anti-nutritional factors, it is a simple and effective way for improving food’s nutritional quality (Aguilera et al., 2015). As evidenced by the fact that certain seeds change in flavour during germination (Kaczmarska et al., 2018). Thus, they showed that throughout the sprouting process, the concentration of volatile organic components named as 2-methylbutanal and dimethyltrisulfite increased that have the impact of increasing the flavour of the seeds. At the same time, the seeds’ sweetness was increased. According to research by (Xu et al., 2019a, b), after 4 days the taste profile of chickpea seeds that had undergone the germination process had changed negatively. It seems that a longer germination period enhances the distinctive flavour of the beans as in the cases of lentils and chickpeas, but in chickpea case, the distinctive
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flavour of the beans diminished and unpleasant odours appeared. After germination, the aroma profiles of lentil and pea were quite similar. According to further study, in chickpea flour, the distinctive flavour of beans faded and unpleasant scents seemed. It is desirable to examine changes in flavour profile during the germination process in order to properly boost the nutritional profile of the seeds without sacrificing their flavour. In general, as the germination period increased, the sensory profile of the grains and legumes that were treated to the procedure became more and more obvious. In the case of chickpeas, lentils, and peas, (Xu et al., 2020a, b) observed that as the length of time the legumes were exposed to the germination method grew, the characteristic aroma of beans became more perceptible. They showed that the concentration of three molecules essential to the development of the sensory profile diminished with increasing germination period. This is due to germination, which encourages the elimination of the flavour components by damaging the endosperm. To decrease the unique odour of beans, antioxidants may be introduced during the germination phase.
10.2.1 Health Benefits of Sprouts Popularity of sprouted seeds in human diet increasing day by day owing to the positive health impact. People are more aware and having lot of information about the health benefits of sprouts intake in their daily diet. This is due to the growing demand of therapeutic (sprouted foods) and functional foods to boost immunity in view of latest coronavirus outbreak (Le et al., 2020). Mostly, germinated legume seeds were consumed as sprouts in the human diet while cereal sprouts were used in fodder as animal diet (Waginger H, 1984). Sprouts contains numerous biologically active components with potential health aids. It comprises of variety of important constituents that helps in preventing various human health issues. Moreover, played vital role in avoidance of different kinds of cancers (Teixeira-Guedes et al., 2019; Ki et al., 2017). Mung beans sprouts contributing in reduction of heat strokes and gastro-intestinal disorders (Tang et al., 2014). Yao et al., 2008 reported mung bean sprouts improves glucose tolerance and contains anti-diabetic potential. They analysed after the consumption of mung bean sprouts by type-2 diabetic mice for 5 weeks, there was reduction in the level of blood glucose, triglycerides and cholesterol. Soybean sprouts reduce the cancer risk and cardiovascular diseases due to health promoting properties (Prakash et al., 2007). Likewise, Nakamura et al. (2001) stated due to the presence of iso-flavonoids, soy bean sprouts have been found to give protection from cancer and cardiovascular disease. Cowpea sprouts helps in decreasing cell proliferation while increasing anti-colorectal cancer (Teixeira-Guedes et al., 2019). The anti-bacterial activity of pea sprouts has been shown against Helicobacter pylori accompanied by gastric tumor (Ho et al., 2006).
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10.3 Food Safety Issues Related with Sprouts Sprouts are possible natural source of wide range of bioactive constituents that have a variety of health benefits in the treatment and prevention of illnesses. Sprouts are mature form of edible seeds with enhanced nutritional value. Soaking is necessary step for making the seed. Edible seeds are raised to germinate physically by soaking in salt water (Aloo et al., 2021; National Advisory Committee on Microbiological Criteria for Foods, 1999). The US FDA has issued various regulations and guidelines and recommendations for good agricultural practices for natural plant products which are useful for sprout developers to avoid possible risks of contamination during processing of sprouts. Because bacteria thrive in damp environments, soaking creates a favorable setting for their growth. As a result, it’s important to replace the soaking water often to avoid the development of germs. While limited soaking does not help the enhancement of phytochemical content in seeds, excessive soaking may promote microbial growth and seed fermentation. In order to ensure optimal soaking, variables such the seed weight/water volume ratio, soaking time, and soaking temperature are tracked. In general, soaking can be carried out for up to 24 h at room temperature depending on the qualities of various seeds. Germs thrive in the warm, moist conditions that are required to develop sprouts. Alfalfa, bean and other sprouts may cause food poisoning from Salmonella if consumed raw or gently cooked. Proper maintenance of sprouts is recommended in the refrigerator at 40 °F or less. Consumption of sprouts is quite common in our daily diet but at the same time ensuring their quality or contamination level is more important in order to avoid food poisoning with Salmonella, Listeria and E.coli. Any form of raw or barely cooked, still crunchy sprouts should not be consumed by the people with lowest or impaired immune systems. Home-grown sprouts are equally dangerous to consume uncooked (Chiriac et al., 2020; Ertop et al., 2018; U.S. EPA, 1993).
10.4 Examples of Common Sprouts Every viable seeds are known to be sprouted such as (a) Pulses (lentils, peas, mung beans, chickpeas, black eyed peas and soybeans) (b) Cereals (wheat, maize, oats, rice, barley and rye and (c) Oilseeds (peanuts, almonds, linseed, sesame and sunflower. Their regular consumption as a part of human diet is increasing day by day. Here, in this chapter we discussed some common sprouts such as black eyed peas, chickpea and pea, their health benefits, nutritional value and applications.
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10.4.1 Black Eyed Peas The Black eyed pea or black eyed bean, southern pea and sometimes called cowpea (Vigna unguiculata) is one of the important legume grown in various parts of India. It is a multipurpose legume with a remarkable nutritional profile. All of the cultivated cowpeas are categorized under the species Vigna unguiculata. This species is divided into 4 cultivera: Unguiculata, Biflora, Sesquipedalis and Textilis. In the Indian Subcontinent, it is also referred to as chawli or lobia in Hindi, alachandalu or bobbarlu in Telugu, alasande in Kannada, karamani payir in Tamil, vanpayar in Malayalam, and barbate in Bengali. It is an annual perennial bean that is grown extensively in the arid parts of Africa and Asia. It is a member of the Fabaceae family. Due to the shade effect of their broad, drooping leaves, cowpeas are noted for their drought-resistant characteristics. It is likewise famous as southern pea or black-eyed pea (Becerra-Tomas et al., 2019; Bielefeld et al., 2020; Wang et al., 2021).
Black eyed pea (Vigna unguiculata)
Commonly cultivated in the arid and semi-arid regions such as Goa, Delhi, Punjab, Haryana, Uttar Pradesh, Karnataka, Kerala, Tamil nadu, Gujarat, Maharastra and Rajasthan. A number of varieties of Black eyed pea are existed like Pusa Phalguni, Amba (V-16) (M), Ramba (V240) (M), Swarna (V-38) (M) etc., still, Goan cowpea, local name Alsando is distinctive and likely being grown by the farmers (Naik et al., 2017). They typically have an eye-like black, brown, or red mark on their body that is quite noticeable. Due to their strong, savoury flavour, black eyed peas are frequently regarded as a cornerstone in both Indian and traditional Southern cuisine. It is used to prepare the traditionally in Punjabi families and is also used to create gravies, soups, and salads. It has a distinct nutty, earthy flavour and a rich creamy flavor. Commonly, popular food as a great vegetarian source of protein, high in fibre and include essential vitamins and minerals. Taste great in salads, tacos, soups, and other foods. In addition to provide a number of health benefits. Consumer may be able to improve their healthy gut flora and reduce cholesterol and blood sugar by increasing diet of these foods.
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10.4.1.1 Nutritional Composition The black eyed pea contains significant amount of protein, carbohydrate, amino acids, water-soluble vitamins, minerals and dietary fibre (Devi et al., 2015). Black- eyed peas are highly nutrient-dense due to the quantity of fibre and protein in each serving. Additionally, they are a great source of a variety of essential micronutrients, including folate, copper, thiamine, and iron. Cowpea seeds, including undeveloped (frozen or cooked), matured (raw or cooked), whole grain, leaves and decorticated grain are important sources of carbohydrates, proteins, lipids, minerals, total dietary fibres and vitamins that remain vital for both human and animal nutrition. Cowpea whole grain protein content lies between 23 and 32%, carbohydrate content between 50 and 60% and fat content less than 1% fat. As compare to cereals and tubers, sprouts contain 2–4% more total protein. It contains an adequate quantity of dietary fibre (soluble and insoluble) as well as phytochemicals, minerals and vitamins (Jose et al., 2014; Kirse & Karklina, 2015; Trehan et al., 2015; Aguilera et al., 2013). Cowpea seeds provide a sizable quantity of dietary protein for masses of people in less developing countries, enhancing the nutritive value of low protein-rich cereals and tuber based staples. Cowpeas come in a variety of sizes and shapes and all of them, including the mature seeds (dry grain), green seeds and green pods are all healthy. They include anti-nutritional components that could be harmful for humans and non-ruminant animals to consume. To minimise or completely eradicate the detrimental effects of anti-nutritional components, several processing steps are followed. Due to its high protein, fibre, and low glycemic index, the cowpea is a very nutritious diet that may also have therapeutic effects (Aloo et al., 2021; National Advisory Committee on Microbiological Criteria for Foods, 1999; Idowu et al., 2020). 10.4.1.2 Health Benefits Because they include protein and soluble fibre, they aid in weight loss. The fibre is slowly absorbed by your body and the protein keeps you feeling fuller for longer. Additionally, soluble fibre enhances intestinal health and prevents constipation; Consume more black eyed peas to help control blood pressure and harmful cholesterol levels, which will benefit your heart health. The amount of folate, zinc, magnesium, vitamins, and protein may rise while eating sprouted beans, making them more nutrient-dense. The starchy endosperm is broken down during the sprouting process, making the beans easier to digest. Promotes Loss of Weight The greatest strategy to lose extra weight is to regularly include chawli in your diet plan because it is high in protein and soluble fibre. Foods high in protein lower ghrelin levels, which is a hormone that makes you feel hungry. Soluble fibre keeps
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you full and helps you manage unwelcome hunger pains while delaying the time until the stomach empties. Additionally, studies have shown that include black-eyed peas in the diet significantly decreased belly fat and maintained weight. In an experimental study, 25 rats were divided into 5 groups and after a week of acclimatization, they were fed with 20% fat as a basal diet and compared to 20% fat enriched diets having 20% whole cowpea powder from the 4 different cowpea cultivars. The end result shows loss in liver weight in rats (Perera et al., 2016). Improves Digestive Health Dietary fibre, a crucial component for preserving the regular operation of the gastrointestinal tract, is found in abundance in chawli. It is commonly recognized that eating a diet high in soluble fibre can alleviate stomach ulcers, acid reflux, haemorrhoids, and regular bowel motions. Additionally, cowpeas work as a prebiotics, promoting the development of the beneficial bacteria in the stomach that help to maintain a healthy microbiome that supports healthy digestion, reduces inflammation, and strengthens the immune system. A randomized control trial runned parallel in a experiment. In one study, children of age 5.5–6.5 months were included and in another study 12–13 months’ age group were enrolled for the randomized intervention for complete 12 months. Children from study 1 and study 2 will consume cowpeas, common beans or standard food up to 6 months. Then, Environmental enteric dysfunction [EED] was scrutinized by quantifying intestinal mRNA for inflammatory messages. It is reported via this experiment that cow pea has the potential benefit of reducing the EED in high risk patients (Trehan et al., 2015). Controls Diabetes Because cowpeas naturally have a lower glycemic index than other legumes and lentils, they help diabetes people keep their blood sugar levels within the usual range. In addition, soluble dietary fibre and protein are excellent for you because they keep you fuller longer, delay stomach emptying, and prevent blood sugar spikes. In another study performed on rats, its results showed that the wild cowpea, white cowpea, african have upturned diabetes-associated dyslipidemia as specified by means of the indexes of cardiovascular disorder (Trehan et al., 2015). Improves Heart Health A nutritious diet that includes cowpeas is a fantastic method to improve cardiac health and lessen the threat of heart ailments. The inherent flavonoid, magnesium, and potassium richness of cowpea maintains the proper operation of the heart muscles and reduces inflammatory indicators. Although the amount of dietary fibre and protein aids in lowering blood pressure, triglyceride levels, and bad cholesterol
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(LDL). In addition, chawli has phytosterol components that support a healthy lipid balance in the body. The hypocholesterolemic activity of cowpeas and isolates of cowpea are reported both in vivo and in vitro. A phenolic compound was reported by Cui et al. (2012); Hachibamba et al. (2013) and Salawu et al. (2014) in cowpea that protects the LDL from the copper induced peroxidation. It is found that Oxidized LDL have a significant role in the Coronary heart disease as it triggers the formation of fatty streaks within the blood vessels. This ultimately will lead to the development of atherosclerotic disease. Hence, cowpea helps in protection against heart diseases. Good for Skin The abundance of protein, zinc, and vitamins like A and C in cowpea stimulates the collagen production and speeds up the growth of new skin cells and skin healing. Due to its enormous antioxidant content, it protects skin cells from oxidative loss brought on by free radicals, reducing wrinkles and fine lines and delaying the onset of age-related changes. Resulting in soft, healthy, and beautiful skin (Ishola & Aramide, 2022). Fighting Infections Chawli has a key role in reducing the risk of developing chronic illnesses since it is sacred by way of a variety of antioxidants, including vitamins (A and C) and polyphenols. Cowpeas can assist eliminate harmful free radicals on a regular basis, preventing the development of cancer cells and preserving good health (Jayathilake et al., 2018). Good for Pregnant Females Chawli has a lot of folate, which helps the body make and maintain red blood cells. This vitamin is critical for both women who are trying to conceive and pregnant women. Because folate is essential in preventing congenital abnormalities in the pregnancy (Osunbitan et al., 2016). Control Hair Loss and Promotes Hair Growth Cowpea, which is endowed with a wealth of nutrients beneficial to hair, acts as a cure for hair loss. A food plan that includes cowpeas helps to improve hair follicles and lessen hair loss. Cowpeas significantly speed up the development of hair. Being abundant in protein, one of the key factors in hair development. Therefore,
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consuming cowpeas on a daily basis can increase their protein content and contribute to the strength and volume of hair (Sowmya Binu 2022). 10.4.1.3 Application of Cowpeas in Food Systems Cowpea Flour There is considerable interest for the fortification of wheat flour with great amount of protein and lysine. It improves the content of essential amino acid of baked food products. Being a good source of protein and lysine, cow pea flour would be a great substitute, particularly in breads. Cowpea Protein Isolate Cow pea protein isolate is made from defatted flour by alkaline extraction. It can be used in different forms such as supplements and ingredients in food processing. Owing to good water absorption and oil absorption properties, it has been utilized as an component in scheming foodstuffs with improved nutritive value. Moreover, tremendous usage in the cheese and meat products as extenders and in baked foods for the purpose of impeding the staling process (Rudra et al., 2016). Akara Akara, known with different names such as Koose or Ata is deep fried and ball shaped fritters. Mostly eat in the form of breakfast or snacks in countries like Nigeria, Ghana and other West African countries. It is processed by grinding decorticated and fully hydrated cowpea cotyledons into a paste form. Addition of water into paste followed by whipping, addition of salt, onion and pepper. Deep-frying until crispier texture and golden brown color appears. Generally, cooked by the lady street hawkers in West Africa (Lowenberg-DeBoer & Ibro, 2008). Reduction of radical scavenging capacity and phenolic content of the product by frying has been reported (Apea-Bah et al., 2017). 10.4.1.4 Sprouting Effect on Nutritional Composition of Black Eyed Pea When water enters into the seed coat, seed swells and sprouting process initiates. During the sprouting process, intake of water diverges. The seeds absorb water through a process known as imbibitions (Nonogaki et al., 2010, Sampath et al., 2008). It has been found that sprouting significantly increased the moisture content of black eyed peas (devi et al., 2015). Also, Uwaegbute et al. (2000) analysed that
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the moisture content of black eyed pea improved from 15.6% to 17.6% after the sprouting process. A gradual increase in the crude protein content (19.15%) was noticed after 28 h of black eyed pea sprouting (Mehta et al., 2007), 8–11% increase in crude protein (Uppal & Bains, 2012). Owing to the depletion of the stored fat that contributes the catabolic activities of seeds, there was decrease in crude fat content during the sprouting process (Onimawo & Asugo, 2004).
Black eyed pea sprouts
It has been found that crude fibre content increased after sprouting. Uppal and Bains (2012) reported 20–24% increase in crude fibre afterward sprouting. A gradual rise in crude fibre content (Sood et al., 2002) and decrease in the carbohydrate content after sprouting has been reported. A 5.6% decrease in carbohydrate was observed by Uppal and Bains (2012) and 2.34% decrease in carbohydrate content after 24 h of sprouting (Jirapa et al., 2001). Sprouting of cow pea increased the calcium content by 9.97%, 5.94% and 7.24% in three different cultivars (Devi et al., 2015). Another study revealed 24–62% increase in calcium content after sprouting. After sprouting, a sudden increase in vitamin C amount of the three cowpea genotypes was strangely important. Anti-nutrients play a vital role, a substantial lessening in anti-nutrient content was noticed after sprouting (Devi et al., 2015). Decrease in phytic content (43.19%) after 24 h of sprouting in cow pea was reported (Uppal & Bains, 2012). some of the anti-nutritional factors like phytic acid and trypsin inhibitor lessened afterwards sprouting of black eyed pea (Devi et al., 2015).
10.4.2 Chick Peas The chickpea (Cicer arietinum) is one of the important legume of family Fabaceae are known for various names such as Bengal gram, Gram, Garbanzo bean Desi chana, Kabuli chana and Chana Numerous varieties of chickpeas are cultivated throughout the world. Commonly cultivated in countries such as India, Pakistan, Turkey, Burma and Ethiopia. But, India got first rank in worldwide chickpea
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production. According to reports, in 2019 India was responsible for about 70% global chickpea production. (FAOSTAT, 2022). Also, known as essential legume crop in India particularly cultivated in Madhya Pradesh, Uttar Pradesh, Haryana, Punjab, Rajasthan and Maharashtra. Chick pea serves as energy and protein source food that contains carbohydrates, fat, dietary fibre, calcium, iron and niacin.
ChickPea (Cicer arietinum)
10.4.2.1 Nutritional Composition chickpea is a rich source of carbohydrates (60–70% and proteins (18–24%), altogether constituting around 80% of the total dry seed mass (Chibbar et al., 2010). According to Yust et al. (2012), globulins account for the majority of the increased protein bioavailability of chickpea protein which is observed to be superior. Contrary to cereals, which are abundant in sulfur comprising amino acids and in which lysine serves as the restrictive amino acid, chickpea seeds are lacking in methionine and cysteine (Sulfur-Containing Amino Acids) while great in lysine and arginine ((Rachwa-Rosiak et al., 2015a, b. Protein efficiency ratio (PER), biological value (BV), net protein utilisation (NPU), and total digestibility (TD) ranges for raw chickpea seeds are 1.2–2.8, 0.520–0.850, 0.556–0.920, and 0.760–0.928, respectively. Aspartic acid, glutamic acid, and arginine were the most abundant amino acids in the Kabuli and Desi chickpea protein isolates. The fraction of amino acids containing sulphur was likewise extremely high in these samples .(Wang et al., 2010). About 41% to 50% of starch amount of chickpea cultivars have been stated. The kabuli chana variety constitute more soluble sugars (Jambunathan & Singh, 1980). Chickpea starch grains are even and oval. The amylose content of pea starch is high, which causes it to undergo greater retrogradation and consequently lower glycemic index, as the starch turn into resistant to enzymatic degradation (Fredriksson et al., 2000). Chickpea starch was found to have a glycemic index of 49.8. The low glycemic index of chickpea starch is owing to their higher amylose content and resistant starch (Foster-Powell et al., 2002; Sandhu & Lim, 2008). Resistant starch has a biological function similar to fiber because it spreads the large intestine deprived of being hydrolyzed or broken down. Moreover, presence of 6% fat make it essential to be portion of vegetarian diets of resource-poor consumers. The fat content in the Bengal gram (raw)seeds varies
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from 2.70% to 6.48% (Kaur & Singh, 2005). Small differences in fat content value 2.05 g/100 g have been reported by various researchers (Shad et al., 2009), 2.90–7.42 g/100 g (Wood & Grusak, 2007) and 3.80–10.20 g/100 g (Singh, 1985). The fatty acid composition of chickpeas is: 66% polyunsaturated fatty acid, 19% monounsaturated fatty acid and saturated fatty acids are about 15%. The most dominating fatty acid present in chickpea is linoleic acid (51.2%) followed by the oleic acid (32.6%). In contrast, the quantity of linoleic acid and oleic acid is lesser in other legumes (linoleic acid: beans, peas and lentils 46.7%, 45.6% and 44.4% respectively and oleic acid 28.1%, 23.2% and 20.9% recpectively) (Wang & Daun, 2004). Despite having little lipid content, chickpeas are high in linoleic and oleic acids, which are essential for good nutrition. Important sterols found in chickpea oil include stigmasterol, campesterol, and sitosterol. The lipid fraction (4.5–6.6%) is made up of carotenoids (46.3 g/100 g of flour) and tocopherols (230.3 mg/100 g of oil), as well as fatty acids such as palmitic (10.8%), oleic (33.5%), linoleic (49.7%), and linolenic (2.4%) (Jukanti et al., 2012; Ryan et al., 2007). The fibre content of varieties such as kabuli and desi altered quantitatively and qualitatively (Singh, 1985). Desi-type chickpea has higher total fiber and insoluble fiber content than Kabul-type chickpea due to thicker seed coat and skin (Rincon et al., 1998). Soluble and insoluble fiber in chickpea is about 1.42% and 27.84% (Khatoon & Prakash, 2004). The soluble and in-soluble dietary fibers content in legume flour is greatly influenced by the processing technique. In germinated pea seeds, a reduction in the proportion of insoluble to soluble fiber was detected, on the other hand quantity of both types of fiber increased. This specified that the rise in soluble fiber was more reflective than insoluble fiber (Martin-Cabrejas et al., 2003). Extrusion cooking also leads to significant conversion of insoluble dietary fibers to soluble due to reasonable dissolution of cell wall polymers (Ralet et al., 1993). In dicots, the most important cell wall polysaccharides are xyloglucans (hemicelluloses) and homogalacturonan and rhamnogalacturonan (pectin) (Tosh & Yada, 2010). Legumes comprised of significant amounts of α-galacto-oligosaccharides, mostly related to the raffinose family, which comprises raffinose, verbascose and stachyose (Guillon & Champ, 2002). Chickpea holds significant minerals like calcium and iron (Murty et al., 2010). It has been reported that Immature green chickpea seeds included 2.2 mg thiamine (100 g)−1 and .7 mg riboflavin (100 g)−1 (Geervani & Devi, 1989). Chickpea seeds also include Ca, Mg, P, and particularly K. Important vitamins like riboflavin, niacin, thiamine, folate, and the precursor to vitamin A, alpha-carotene, are all found in abundance in chickpeas (Hirdyani, 2014) When combined with other pulses and cereals, chickpeas may have positive effects on a number of serious human ailments, including cardiovascular disease (CVD), type 2 diabetes, digestive disorders, and several malignancies. (Jukanti et al., 2012). Daily requirements for iron and zinc can be encountered by eating 100 g of chickpeas, and eating 200 g of chickpeas can encounter the daily requirement for magnesium (WHO, 2004). Desi chickpeas have a higher calcium content than kabuli chickpeas. Also, contains significant quantities of Vitamin E (Tocopherol) and
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Vitamin B9 (Folic Acid) as 11.2 and 206.5 mg/100 g in desi and 12.9 in and 299.0 mg/g in kabuli chickpeas. (Ciftci et al., 2010). Desi Chickpeas too comprises lesser amounts of vitamin B complex, primarily vitamin B2 (also called as riboflavin) 0.21 mg/100 g, B5 (also called as pantothenic acid)1.01 mg/100 g, B6 (also called as pyridoxine) 0.30 mg/100 g whereas 0.26, 1.02, 0.38 mg/100 g respectively in the Kabuli chickpeas (Lebiedzinska & Szefer, 2006; Wang & Daun, 2004). The amount of Iron and Zinc varies in the range of 4.9–5.0 mg/100 g and 2.0–2.7 mg/100 g, respectively (Hemalatha et al., 2007). The proportion of micronutrients in chickpeas are highly influenced by its genotype. It was reported that selenium concentration lies in between the range of 15.3 and 56.3 μg/100 g in various varieties of chickpeas grown in North America. (Thavarajah, 2012). Some of the health benefits of chickpeas are believed to be due to the existence of bioactive compounds. The most important phytochemicals in chickpeas include flavonoids, carotenoids, phenolic acids, stilbenes and lignans (Xu & Chang, 2007). Antioxidants are found to be in higher amounts in Desi chickpeas than in the Kabuli variety (Segev et al., 2010). Most of the flavonoids and phenols contained in chickpeas are mainly intense in the seed coat and the flavonoid activity, total phenolic activity, antioxidant activity content are 11, 13, 31 times, respectively higher in the brown desi type than in the creamish kabuli type (Segev et al., 2010). Chickpea seeds are richer in carotenoids (mainly xanthophylls, cryptoxanthins and beta- carotene). Carotenoids are very important because they rise the bioavailability of iron in the human body by act as an iron absorption enhancer (Welch, 2002). Chickpeas, like many other legumes, contain certain anti-nutritional factors that can affect their nutritional value and digestibility. Here are some common anti- nutritional factors found in chickpeas (Table 10.1): Table 10.1 Anti-nutritional factors in chickpea Phytic acid
Chickpeas contain phytic acid, also known as phytate, storage form of phosphorus in plants. Phytic acid can bind to minerals such as iron, zinc, calcium, and magnesium, reducing their bioavailability and inhibiting their absorption in the body Protease Chickpeas contain protease inhibitors, compounds that inhibit inhibitors the activity of digestive enzymes called proteases. Protease inhibitors can interfere with protein digestion, potentially reducing the availability of dietary proteins Lectins Lectins are proteins that can bind to the lining of the digestive tract, interfering with nutrient absorption. Some people may be sensitive to specific lectins found in chickpeas, such as agglutinins, which can cause digestive discomfort Oligosaccharides Chickpeas contain complex carbohydrates called oligosaccharides, such as raffinose and stachyose. These carbohydrates are not easily digested by humans due to the lack of specific enzymes in our digestive system. As a result, they can cause flatulence and digestive discomfort in some individuals
Zhang et al. (2022)
Thakur et al. (2019)
Popova and Mihayloya (2019) Li et al. (2022)
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While these anti-nutritional factors can have adverse effects, there are ways to mitigate their impact. Soaking and cooking chickpeas can help reduce the levels of phytic acid and protease inhibitors. Additionally, sprouting or fermenting chickpeas can further enhance their nutritional value by degrading somehow the anti-nutritional factors. These methods can increase the digestibility and bioavailability of the various nutrients present in chickpeas (Samtiya et al., 2020). 10.4.2.2 Application of Chickpea in Food Systems Compared to another pulses, chickpea is consumed in a variety of forms and in a variety of products (Joshi et al., 2001). Since ancient times, chickpeas have been used in the manufacture of several traditional culinary preparations. It is used in the production of a variety of dishes as a foundation ingredient or an additive. Chickpeas are eaten as a vegetable with meals when they are picked when they are in the green (raw) stage, which is 10–15 days earlier they reach maturity. After going through various processing steps as being soaked, boiled, germinated, or roasted chickpea, it may be eaten as whole grain. The most typical way to consume chickpeas is as “chana dhal,” or split chickpeas (Jambunathan & Singh, 1989). The flour made from the chickpea dhal is known as “besan” and is used to make a variety of snacks, desserts, bakery goods, etc. It can be used alone or in blend with other cereal and pulse flours (Wood & Malcolmson, 2011). Studies have previously shown that eating chickpeas has a number of physiological advantages and lowers the chance of developing chronic illnesses (Duranti, 2006; Murty et al., 2010; Wood & Grusak, 2007). Chickpeas are a functional food because they are high in protein, have dietary fibre, and have antioxidant and anti- inflammatory effects. They also have a low glycemic index (Crujeiras et al., 2007; Fredriksson et al., 2000). It is detailed how chickpeas are used in the food chain in various ways. Whole Grain Chickpeas that are still green are frequently eaten as vegetables, and grains are picked 10–15 days before they reach full maturity. Grain that has reached full maturity and ripeness is collected for storage and for further processing purpose. Before being consumed, dry chickpea seeds are often put through several technical procedures. Most processing methods, including germination, roasting, and fermentation, begin with soaking. It enables grain moisture content to rise and cell water absorption, cutting down on cooking time period (Prasad et al., 2010; Kumar et al., 2018). Chickpeas that have been cooked and then soaked are eaten raw or in a salad. Antinutritional elements can also be removed by soaking. Additionally, by enhancing the heat transfer phenomena when cooking, it shortens the cooking time (Wang et al., 2019). During soaking at 10–60 °C, chickpea seeds weight is observed to rise
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by 1.76–2.12 times (Kaur & Prasad, 2021a). Additionally, it has been discovered that chickpea seeds may germinate at a soaking temperature of 30 °C (Unpublished research data). Various ways that chickpeas are frequently consumed. To make “hummus,” soaked Kabuli chickpea are mashed into a paste and then combined with oil and seasonings. Uses for hummus as a spread or dip (Wallace et al., 2016). By allowing wet chickpea seeds to germinate, chickpea sprouts are created. It is frequently eaten as chickpea salad because to its higher nutritious value and mouthwatering flavour. Germination raises the bioavailability of minerals, decreases the anti-nutritional constituents and rises the digestibility of proteins (El-Adawy, 2002). Due to enzymatic activity during the germination phase, the oligosaccharides, primarily stachyose and raffinose, are reduced (Xu et al., 2019a, b). In India, roasted chickpeas—either hulled or unhulled—are frequently eaten as a snack dish (Sahu et al., 2014). Roasted chickpeas are covered with jaggery or other spices to make them more palatable. (Raza et al., 2019; Kumar & Prasad, 2017, 2018; Kumar et al., 2016; Sharma & Prasad, 2016; Sharma et al., 2015) Roasting is a heat treatment that improves texture, colour, and flavour. The primary cause of the colour and flavour changes that occur during roasting is the transformation of carbohydrates into dextrins which then interact with amino acids (Gahlawat & Sehgal, 1992; Mariod et al., 2012). The desi chickpea’s seed coat is removed, and the cotyledon is separated into two halves to create chickpea splits. It is frequently referred to as “chana dhal.” in India. The majority of chickpeas consumed are consumed as “dhal” or “split” (Mangaraj et al., 2005). It is often eaten with rice or chapatti and boiled in water with additional spices. As a result, the amalgamation of cereal and pulses offers a composition of amino acids that is balanced (Young & Pellett, 1994) (Fig. 10.1). Chickpea Flour Dehulled desi chickpeas are used to make the flour known as “besan.” It serves as a foundational ingredient in the creation of culinary items. Chapati is frequently made with a besan and wheat flour mixture (Malhotra et al., 1987). Besan is used to make a variety of traditional foods in India, including bhujia, dhokla, pakoras, boondi, and sweets, by blending it into a batter, fine paste or dough (Wood & Malcolmson, 2011). Roasted Chickpea Flour (Sattu) The old Indian cuisine dish known as “Chane ka Sattu” is made mostly from roasted Bengal gramme. It may also be made with a blend of flours made from roasted grains and pulses, which will improve its nutritional profile. Popular summer beverage “Sattu” is extremely popular in rural India (Kaur & Prasad, 2021b). It may be advantageous to fortify “Sattu” with vitamins and minerals in accordance with the
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Fig. 10.1 Uses of chickpea in different forms
day-to-day recommended consumption to improve its nutritional profile and address the issue of undernutrition (Prasad, 2009). The addition of cereal flour to roasted Bengal gram flour increases the protein content and balances the amino acid profile. Its additional fortification with vitamins and minerals may help people maintain a healthy diet. With the danger of protein and energy malnutrition, babies in the translational phase and those who have trouble chewing food may benefit from this in terms of delivering adequate nutrition. It is advantageous for diabetes individuals because of its low glycemic index. 10.4.2.3 Germination Effect on Nutritional Composition of Chickpea As we previously stated, enhancing the nutritional value of meals is the main reason that germinated seeds or the flour formed from them is used in the recipes of various foodstuffs. Along with the physical changes, the nutritional makeup of the seeds also changes during germination. Studies have shown, for instance, that the lipid content of the seeds is impacted by the germination process. Thus, (Xu et al., 2020a, b) came to the conclusion that the reduction in fat content may have been caused by the rise in lipolytic activity throughout the germination period. As a result, lipid
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molecules are broken down to make sure seed development. Whether or not the amount of fat is lowered depends on the species that is in the germination period. Numerous studies indicate that during the germination stage, the ash content of rice and amaranth increased. This resulted from a diminution in the amount of soluble solids (such as starch and different sugars). Germination doesn’t appear to significantly alter the quantity of carbohydrates, but it does seem to decrease the amount of amylose and increase the amount of total sugar. Because the germination process increases the nutritive value of the seeds, they can be successfully used in baked goods (Cornejo et al., 2019). The maximum antioxidant capacity was demonstrated by a 32 h germination time. After germination for 8 h, the antioxidant capacity began to rise. There has lately been growing interest in chickpeas since they are a fantastic source of proteins, fibre, carbohydrates, and minerals and help maintain a balanced diet. For vegans, chickpeas are a fantastic source of protein. Furthermore, it contains less allergen. At the same time, chickpeas may be seen as a soy alternative. Studies have shown that chickpeas have an in vitro digestibility that is between 48% and 89.01% greater than that of beans and soybeans (Rachwa-Rosiak et al., 2015a, b). It has been shown that germination enhances the total content of flavonoids, polyphenolic compounds having antioxidant action, in the case of chickpeas. The total flavonoid content rises from 0.22 to 0.42 g/kg following germination at a germination temperature of 30 °C. At germination temperatures greater than 10 °C, the total flavonoid concentration was 0.38 g/ kg. Therefore, it is possible to say that the germination temperature influences how much the flavonoid amount grows. The seed’s response to numerous biotic and abiotic stressors is crucial to its development. However, germination-related changes in the total flavonoid concentration are unique for each species of seed. This transformation takes place based on the physical characteristics of the seed, its class, and its hardness (Kigel et al., 2015). According to research done thus far, the amount of proteins, lipids, fibre, ash, and carbohydrates in chickpeas is not significantly affected by the process of germination. However, this procedure results in more ascorbic acid being generated. Its concentration rises from 1.9 mg/100 g to 9.4 mg/100 g to 15.6 mg/100 g following a 24 h germination period (after a germination period of 48 h). A rise in a few important amino acids was also seen after germination. For instance, the amounts of threosine, lysine, leucine, valine, and isoleucine increased later 24 h of germination, whereas for some of them, their levels decreased after 48 h of germination. Of all the amino acids, lysine had the greatest increase (Fernandez & Berry, 1988). However, studies have shown that chickpeas’ nutritious content can only be increased by their germination under optimum conditions. Monitoring the temperature and germination period is crucial for achieving the necessary nutritional enhancement. A research study by Dominguez-Arispuro et al. (2018) found that 27.5–35 °C and 125–240 h were the optimal temperatures for chickpea germination. The nutritional components (polyphenol content, total flavonoid content, and antioxidant activity) showed the highest values for the germination process parameters of 33.7 °C and 171 h. It has been proven that under these conditions, chickpeas have a much larger amount of nutritional fibre. Additionally,
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it was shown that ferulic and ellagic acids, both have antioxidant characteristics, considerably increased during germination. The protein content considerably increased as a result of the germination process which took place for 171 h at 33.7 °C (Domnguez-Arispuro et al., 2018). The rise in protein content has been associated with the damage of dry matter during the seed respiration, germination, mainly in the form of carbohydrates. As a result of lipids being utilized as source of energy during the germination process, a decline in lipid content was also detected. 10.4.2.4 Sprouting Effect on Nutritional Composition of Chickpea Sprouting is a complex metabolic process that increases the bioavailability, affects the palatability and digestibility of nutrients. Conversely, sprouting effect will depend on the variety of legumes, conditions and time duration of sprouting process. Sprouting effected the moisture content, sprouting time significantly influenced the moisture content with the increase in passage of sprouting time period. Moreover, after 120 h sprouting time period, the mean value of moisture content increased to 62.67% in sprouting seeds. Moisture contents of sprouts were significantly (p 1000 mV), and a high concentration of dissolved oxygen. A non-membrane electrolytic cell electrolyzes diluted HCl solution or NaCl to create electrolyzed water slightly acidic with a pH range of 5.0 to 6.5 and strong hypochlorous acid (C. Zhang et al., 2016). To lessen or get rid of pathogens, fungus, spores, and biofilms, both forms of EO water have been used extensively in a variety of areas, including produce, meat products, and seafood. When EO waters along with ACC of 30 to 50 mg/L was sprayed on seed sprouts, the number of aerobic bacteria, mold, and yeast was reduced by 1.39–1.58 logs, and the gross weight, length, and dry weight of the sprouts were all improved. To overturn the naturally microbes on sprouts, EO water with lower ACC and slightly acidic pH was proven best to control microbiota during seed germination and sprouting (C. Zhang et al., 2016). Fresh fruit and vegetable microbial contamination has been thoroughly researched using the non-thermal preservation technique known as cold plasma therapy. Microorganisms are inactivated by cold plasma due to damage to the microbial cell
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membrane caused by radicals, ultraviolet (UV) photons, electrons, and charged plasma particles. Additionally, membranes may be directly bombarded and harmed by electrons, ions, and free radicals, which can result in surface lesions (etching) and the death of these cells. Inside of Salmonella typhimurium (Oh et al., 2017). Despite being extensively investigated and used as new plant meals rich in bioactive compounds, sprouts from the Brassicaceae family lack information on the stability of their phytochemicals over time and the microbial flora they contain. Sprouts should be stored at a temperature of 0 to 2 °C, however some studies have found that more than 40% of the goods kept in supermarket under refrigeration (7 °C). Here, sprouts are treated as fresh goods and consumed as such. In order to assess plant foods as ideal content of phytochemicals and health-consciousness consumers, we looked at the microbial contents along with the contents of isothiocyanates, glucosinolates, and phenolic compounds of sprouts after collection and after 7 or 14 d of storage at 5 °C, which is typically used in normal household refrigeration (Baenas et al., 2017).
16.9 Conclusion Mustard and radish sprouts are safe foods from the perspective of their microbiological and phytochemical content. Furthermore, this finding showed that these sprouts are considered best from standpoint of nutrition. Additionally, this study, makes a scientific contribution to the field of food safety of significant interest for sprout consumers and producers.
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Chapter 17
General Overview of Composition, Use in Human Nutrition, Process of Sprouting, Change in Composition During Sprouting, Parameters Affecting Nutritional Quality During Sprouting, Benefits of Sprouts, Nutritional Values and Food Safety Issues of Allium Sprouts Bindu Bazaria and Neeraj
17.1 Introduction Traditional systems of medicine, contemporary medications, nutraceuticals, dietary supplements, folk remedies, pharmaceutical intermediates, and chemical entities used in the production of synthetic pharmaceuticals all receive their drugs from medicinal plants. Medicinal plants are the richest bio-resource of drugs because they contain active ingredients, medicinal plants can be of use in the treatment of a variety of human illnesses (Doss, 2009). Plants in their whole, including their leaves, vegetables, roots, and seeds, contain phytochemicals. The genus Allium members like garlic, chives, onions, leeks and scallions also possess health improving compounds. These are rich in the sulphur compounds that are responsible for the therapeutic effects. The cloves of garlic which have the shape of teardrops and are covered in dry skin-like sheets, are combined to form the bulb. Sprouting has been increasingly popular in recent years. When compared to consuming mature vegetables, the nutritional advantages of sprouts are far more concentrated, which is helping to drive their meteoric rise in popularity (Naji et al., 2017). Most people eat sprouts uncooked since they are low in calories, high in fibre, enzymes, protein, and a variety of other important micro-nutrients. When B. Bazaria (*) Department of Food & Nutrition and Food Technology, Lady Irwin College (University of Delhi), New Delhi, India e-mail: [email protected] Neeraj Department of Agriculture, Jharkhand Rai University, Ranchi, India © The Author(s), under exclusive license to Springer Nature Switzerland AG 2023 I. Majid et al. (eds.), Advances in Plant Sprouts, https://doi.org/10.1007/978-3-031-40916-5_17
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included in our diet on a regular basis, sprouts are an excellent source of all of the nutrients and vitamins that are necessary. The term “sprouts” (Regulation (EC) No 208/2013) refers to “the product acquired from the germination of seeds and their growth in water or another medium, collected before the formation of real leaves and which is meant to be eaten whole, including the seed.” Sprouting is a physiological phenomenon that occurs after the dormancy of a plant has been broken, and it presents a significant challenge in the context of the preservation of vegetables. The situation known as sprouting is characterized by the stem apex of any vegetable exhibiting signals that it may soon break through the plant’s neck. The sprouts are an excellent source of protein, vitamins, and minerals, and they also include essential components that are crucial for preserving one’s health, such as glucosinolates, phenolic, and selenium. Sprouts are a great addition to any diet. The incorporation of water into the dormant seed at the beginning of the germination process is followed by the formation of the embryonic axis at the completion of the process. When compared to mature plants, sprouted vegetables have much higher amounts of many types of bioactive chemicals, including flavonoids, polyphenols, glucosinolates, and anthocyanins (Hanlon & Barnes, 2011). According to the literature, the amount of storage loss in vegetables can range anywhere from 50 to 90 percent, depending on the genotype and the circumstances of storage. On the other hand, sprouting has been employed extensively as an economical, productive, and simple technique that is handy for expanding the nutritional and functional potential of cereals, pseudo cereals, cruciferous vegetables, and legumes (Manchali et al., 2012). It has been discovered that sprouting is connected with the formation of phytochemical components such as glucosinolates, which are natural antioxidants that have a key role in the prevention of cancer and may thus be used for the development of functional foods (Swieca & Gawlik-Dziki, 2015). The nutritional concentration of the sprouts is maintained at a very high level despite the fact that they are ingested at the beginning of the sprouting phase. During the process of germination, the amount of antinutritive materials such as trypsin inhibitors, phytic acid, pentosan, and tannin is reduced. After germination, compounds with health-maintaining effects and phytochemical properties such as natural antioxidants and glucosinolates are produced, which have the potential to play a significant role in the prevention of cancer as well as other diseases. Therefore, the process of germination has the potential to result in the production of functional foods that have a beneficial impact on the human body and that contribute to the preservation of health (Sangronis & Machado, 2007). According to Reed et al. (2018), this is one of the reasons why sprouts are regarded to be “functional foods.” which in addition to their regular nutritional content, possess qualities that either promote health or prevent disease. Garlic, which is a member of the Liliaceae family, is said to be one of the first therapeutic herbs that was ever discovered. Garlic and the many preparations of garlic have seen considerable use in the field of health benefits, which has led to an abundance of study papers being published in only the past ten years. The fresh bulb has sulfur-containing chemicals alliin, allicin, and volatile oils inside it. Allinase, an
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enzyme, is responsible for the transformation of the odourless chemical alliin into the pungent compound allicin when the garlic clove is crushed. Garlic gets its very strong odour from a compound called allicin. It includes vitamins, minerals and trace elements. On the other hand, garlic is a herbal medicine that is used for the prevention and treatment of many diseases such as cold and flu symptoms through the enhancement of the immune system. Garlic also exhibits anticancer, antioxidant, anti-inflammatory, antimicrobial, antithrombotic, hypocholesterolemic, and hypoglycemic activities. Garlic has been shown to lower blood pressure, cholesterol levels, and blood sugar levels (Thomson et al., 2016). When garlic sprouts, both its nutritional value and commercial viability decrease, and the bulb undergoes remarkable physiological changes. The onion, scientifically known as Allium cepa L., is one of the first known bulb crops and is typically cultivated as an annual all over the world (Brewster, 2008). It is often considered to have originated in Central Asia but now India is home to one of the most significant commercial vegetable cropping operations in the world. It is highly prized for the peculiar and pungent flavour it imparts, and it is an indispensable component in the cuisine of a variety of places. Epidemiological research has shown that eating onions relates to a lower risk of coronary heart disease (Selvaraj, 1976). Flavonoids, such as quercetin, are thought to be a main contributor for this association (Hamauzu et al., 2011). Onion is a rich source of phenols, flavonols (quercetin, isorhamnetin, and kaempferol, and their glycosides), and anthocyanins (cyanidin and peonin), the content of which is dependent on the geographical location, cultivar, climatic circumstances, and storage conditions (Ramos et al., 2006). The anti-asthmatic activity, anti-fungal and anti-bacterial activities have been attributed to the phenolic components, which have been identified as the active components in the compound (Insani et al., 2016). Sprouting, a natural part of the onion crop life cycle is characterized by a rise in the activity of enzymes connected to phenolic metabolism, such as phenylalanine ammonia-lyase and peroxidase (Benkeblia, 2000). The onion bulb is a living system that functions as a natural food store for the plant. This natural food store goes through a natural biological process called sprouting, which is a key contribution to the overall amount of postharvest storage losses. Onions that are allowed to sprout possess an increase in their total protein content, ascorbic acid content, anthocyanins content, total phenol content, total flavonoid content, and antioxidant activity. Sharma et al. (2015) has reported that sprouting onion samples dominate a significant amount of phenolics, as well as 4 total quercetin and quercetin-4′-O-monoglucoside. Chives are said to have originated in Siberia and then spread to America from that region. Chives are members of the onion family and grow in dense clumps that resemble grass. They may reach heights of up to 20 centimetres (eight inches) when grown from bulbs. They have a pungent odour, can withstand harsh conditions, and develop clusters of long, onion-like green leaves that bloom in the spring. The leaves are dark green, erect, and hollow, and they end in pointed apexes. The blooms are dense spheres that range in colour from pink to purple and are composed of a large number of smaller flowers. The bulbs have an oval form and are frequently
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seen grouped together. Lutein, zeaxanthin, and beta-carotene are also present in it (Wang et al., 2011). The chives have been found to contain many types of diallyl sulphides, including diallyl monosulfide, diallyl disulfide, diallyl trisulfide, and diallyl tetrasulfide. The flavonoid glycosides in chives are quercetin glucoside, isorhamnetin glucoside, and kaempferol glucoside (Parvu et al., 2010). Chives have been shown to be effective as antibacterial agents, as well as antithrombotic agents, anticancer agents, hypolipidemic agents, antiarthritic agents, and hypoglycemic agents.
17.2 Process of Sprouting The life cycle of an allium species like onion may be broken down into three distinct stages: rest, dormancy, and renewal (sprouting). The dormancy does not result in a complete cessation of metabolic activity of any vegetable. The rate of respiration, which is widely believed to be a good measure of metabolic activity, is typically considered to be relatively low during dormancy in the bulbs of allium, but it rises after sprouting (Benkeblia et al., 2002). Using shallots and garlic as an example, it has been shown that the garlic bulb must go through a lengthy period of dormancy (four to six months) before it can begin to sprout (Fig. 17.1). When the sprout emerges from the clove, the dormancy period for garlic comes to an end (Qiu & Zhang, 2016). The clove gradually emerges from its dormant state and enters the sprouting stage because of a process that is governed by hormonal variables (Arguello et al., 1991) during post-harvest storage. This process causes the clove to reach the sprouting stage. The subsequent stage sees the sprout begin its process of emanation from the clove. The length of time spent in each of these phases varies greatly depending on the type of garlic cultivar (Burba, 1993). The production of sprouts for commercial use is highly mechanized, and in order for a farmer to make a profit off of a smaller scale of production, they will need to charge higher prices. The Food and Drug Administration (FDA) mandates that facilities that produce sprouts must establish a Hazard Analysis and Critical Control Point (HACCP) plan, one of the components of which is adhering to Good Manufacturing Practices (GMPs). The transfer of water and metabolites from the scales to the base plate marks the beginning of the sprouting process. The process of soaking, draining, and then washing seeds at predetermined intervals until they germinate, also known as sprouting (Table 17.1), and the narrow green colored shoots which emerge out are referred to as sprouts (Anonymous, 2009) (Fig. 17.2). The processes and pieces of machinery used for the cultivation of sprouts differ from one facility to the next. Although commercial storage life and market value of the allium species continue until there is apparent sprout development and roots, dormancy is said to end when the inner sprouts begin to grow, as stated by Abdalla and Mann (1963).
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Fig. 17.1 Growth stage of shallot (a–d) and garlic (e–h): (a, e) adventitious bulb, (b, f) shoot, (c, g) young leaf. (Licensed from: https://creativecommons.org/licenses/by/3.0)
The sprouts grow from this base plate, and the redistribution of phytonutrients is what causes the development of new cells and cell elongation. The meristem mitotic activity increases prior to harvest and throughout sprout growth (7–10 cm), and it decreases noticeably during dormancy. Xihong et al. (2010) studied the effect of various packaging environments (storage at 4 °C for 15 days) on fresh-cut garlic sprouts and discovered that the fresh cut garlic sprouts stored in the Polyvinyl Chloride (PVC) plastic bag had the best quality (Fig. 17.2). To assess the inner sprout growth a dormancy overcoming visual index (DVI) (Burba, 1993) is used which is the percentage ratio between the total length of the
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Table 17.1 General process of vegetable’s sprouting Steps 1. Crop selection 2. Facilities and processing
Details Sprout vegetables: Crops include arugula, broccoli, cabbage, garlic, onions, peas, pumpkin, and radish. Sprouts are produced by placing vegetables in a warm, moist environment until they have germinated to the desired size. Each crop will have its own temperature and soak time requirements, but many germinate in three to seven days. Facilities for production need to be equipped so that air and water temperature can be controlled. Supplemental lights may be required for sprouts in which green color (chlorophyll) development is in demand; however, greening is not desirable for all sprouts. 3. Pest Treatment with calcium hypochlorite, following strict sanitation protocols management during production, and proper storage of the harvested sprouts. Using chlorinated or ozonated irrigation water may also be helpful in reducing the incidence of seed decay. 4. Harvest and Once sprouts have reached the desired size they are rinsed; in some cases, storage remnants of the seed coat are also rinsed off. Excess water is removed using a centrifuge or similar equipment. 5. Packaging Sprouts are packaged for retail sales in clear clamshell containers. Immediate and continued refrigeration is essential for maintaining product quality from harvest to end-market Fig. 17.2 Sprouted onion
sprout and that of the storage leaf, measured in clove longitudinal incisions (Fig. 17.3). Every 30 days (about 4 and a half weeks) following harvest, these measures can be made on samples that each include 10 sprouts. After 90 days of storage, garlic microbulblets are physiologically mature and capable of sprouting (Arguello et al., 2001). A developed, greenish bud is thought to be growing.
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Fig. 17.3 Sprout obtained from a garlic bulb
17.3 Changes in Composition During Sprouting The start of sprouting begins at the same time that the ATP level in sprout leaves increases, which occurs 8 weeks after harvest; however, in the stem plate, it happens sooner while it takes longer in the inner scale. Therefore, it is hypothesized that just before sprout growth begins, the bulb’s center experiences a brief burst of respiration that later spreads to the rest of the bulb (Hussien & Gebhard, 2007). Allium species‘quality and weight loss are key characteristics that are influenced by the coexistence of respiration and desiccation processes. After 10 weeks, onion bulbs loss 50% of their initial weight, according to Sharma et al’s (2014) observations. Up until the end of the fourth week, when visible signs of sprouting and decay begin to appear, the bulbs become whole and healthy. Intense sprouting takes place from the fourth to the eighth week, and at the same time, signs of decay also become more obvious. Onion bulbs lose their spherical shape and shrink to about half their original size by the tenth week, when the decay process reaches its peak. Over a period of storage of several months, the lipids of allium (garlic & onion in particular) sprouts undergo a series of significant compositional changes; these changes are connected to the sprout growth inside the cloves after dormancy. As a result, sprouting onion bulbs breathe more quickly than dormant onion bulbs (Benkeblia et al., 2002). Allium sprouts display a significant range in secondary metabolites like flavanols and amino acids like phenylalanine. Total phenolic and flavonoid show changes after post-storage in ambient conditions, expressed as g GAE/g FW and g Q/g FW, respectively. The 8th week of post-storage observes the highest level of total quercetin in onion sprouts (3.209 0.350 mol/g FW), along with intense sprouting. Additionally, the actual quercetin concentration affects the onion waste’s potential for recovery. Overall, there is a general tendency for the concentration of total
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quercetin and its constituents to rise during the first few weeks of storage and then fall during the final few weeks of post-storage time up until the start of onion sprouting. Up until the sixth week after storage, the phenylalanine content is found to slightly increase in onion bulbs. Inducing a plant defence response, cell injury (which lowers the amount of quercetin-producing units), and the hydrolysis of quercetin conjugates have all been reported to cause changes in chemical composition during post-storage under ambient circumstances (Chung et al., 2011). At both 4 and 21 °C, anthocyanin exhibits an increasing trend between 14 and 42 days after harvesting before beginning to decline towards the end of storage (when sprouting starts). The concentrations of glucose, fructose, and sucrose are reported to have changed after the allium sprouts storage. There are conflicting reports on how much sugar varies amongst onion bulbs at different stages of sprouting, notably between the end of dormancy and this stage. The first six weeks of storage report an increase in total carbohydrates, but after that a gradual decline at an average rate of 1386 g/g FW per week has been reported (Benkeblia et al., 2005). Several researchers have noted a decrease in the sugar content in sprouted allium species, particularly in onions. Chope et al. (2007) has hypothesized in their studies that sugars are metabolized to provide energy for the expanding sprout and that the decline in sugar concentration correlated well with an increase in sprout length. The respiration rates are generally found to be positively correlated with physiological activity (Öpik & Rolfe, 2005). Onion bulbs that are sprouting breathe more quickly than onion bulbs that are dormant (Benkeblia et al., 2002). Although their rate of CO2 production is relatively high immediately following harvest and drying, the rate of CO2 production in “Copra F1” bulbs may also fit into this pattern eight weeks after harvest (Fig. 17.4). Internal sprouting of onion bulbs has been reported by Sharma et al. (2014) to occur during the first week of storage and visible sprouting signs to appear after the fourth week. According to the findings, sprouting times
Fig. 17.4 Changes in sucrose synthase activity of sprouted onion. (Variety: Copra F1) parts
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and the pattern of changes in carbohydrates in garlic cloves stored at 4 and 21 °C are comparable (K. Mashayekhi et al., 2015). According to Kevers et al. (2007), most fruits and vegetables stored in the days following market purchase show an increase in total antioxidant activity, which is also brought on by an increase in phenolic compounds. In the case of onions, the antioxidant activity grow steadily over the course of storage (10 times higher after 23 days after market purchase). According to Benkeblia and Shiomi (2004), total phenolic content slightly increase during the first five weeks of storage and then slightly decrease after seven weeks, when internal sprouting starts. These findings show an increase in the content of querecetin and its glucosides during post storage, suggesting the possible use of sprouted and decayed onion as a source of quercetin and its glucosides.
17.4 Parameters Affecting Nutritional Quality During Sprouting Genotype, environmental conditions that the mother plant experienced, and germination conditions are the main variables affecting sprout composition. The chemical makeup of the bulbs is influenced by external factors in addition to metabolic processes. The action of biological elements like pathogens is also very important (Mark et al., 2002). The chemical makeup of the bulbs can also be affected by interactions with atmospheric air and moisture. According to earlier reports, garlic dormancy is broken by being stored at low temperatures (Vazquez-Barrios et al., 2006). According to Arguello et al. (2001), the process of garlic sprouting starts by cold treatment at 4 °C and Gibberellic Acid (GA3). Wareing and Sannders (1971) hypothesis states that gibberellins (GAs), cytokinins, and the inhibitory hormone abscisic acid work antagonistically to control dormancy in plants (ABA). In onion bulbs or related species, many researchers looked into the relationship between sprouting and changes in endogenous growth substances (auxin, GAs, cytokinins, and ABA) levels. The hydrolytic enzymes result from the highly endogenous GA-like substances that affect the breakdown of starch reserve in the bulbs. Simple sugars are made available for the growth and development of the roots and shoots after starch is broken down. Sprouting occurrs less frequently at lower temperatures than at 21 °C because at higher temperatures, complex oligosaccharides are converted into soluble sugars, which serve as a source for the regrowth of nesting buds before they emerge as sprouts (Hurst et al., 1985). Despite being more expensive than cold or ambient storage, CA storage increases the crop’s worth by lengthening the product’s shelf life. Temperature, carbon dioxide, and oxygen concentrations in the storage environment must all be kept under control in order to safely store CA (Gubb & MacTavish, 2002). Upon returning to the normal temperature range (18–25 °C), the onion bulb undergoes a number of internal changes that prepare it for sprouting. Low-oxygen storage delays weight
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loss, reduces the chance of neck rot, and prevents sprouting. However, severely low oxygen concentrations (0.7%) can cause significant rates of sprouting following removal from storage, as well as unpleasant odours and tissue degradation (Chope et al., 2007).
17.5 Benefits & Nutritional Values of Allium Sprouts Sprouts are a potential natural source of a wide range of bioactive substances that have a variety of positive health effects in the treatment and prevention of diseases. In comparison to raw pastes, sprouted onion pastes are found to retain quality characteristics with fewer colour changes, higher total phenolic and flavonoid content, antioxidant activity, and microbial stability (Majid & Nanda, 2017). The physical and flowability characteristics of onion powders made from sprouted onion varieties are found to be superior to those made from the corresponding raw onion varieties. The microstructural change, larger mean particle diameter, lower values of Carr index, cake strength, and other flow parameters found in the powders from sprouted onion varieties are evidence of their improved flowability for the design of processing equipment for efficient handling and storage. The therapeutic potential of sprouted plant extracts against oxidative damage has made the antioxidant activity of these extracts a major area of study. Sprouts contain a variety of phenolic and nonphenolic compounds that have been found to have antioxidant properties. The activity of ascorbic acid in sprouts has been described (Hamilton & VanderStoep, 1979). As a result, sprouts are gradually being recognized for their useful qualities. Cancer has recently become a more pressing global health issue because of exposure to toxic substances. According to the Centers for Disease Control and Prevention (US), 599,274 deaths in the US occurred specifically due to cancer in 2018 (US, 2005). Scientists have investigated the function of plant sprouts in the treatment of cancer due to the high mortality rate of cancer patients. The anticancer phytochemicals 3-terpene derivatives, 5-flavonoid, −carotene, and lutein have been found in alfalfa, making sprouted seeds important targets in cancer chemoprevention and therapy. A group of metabolic diseases known as diabetes mellitus are distinguished by high blood sugar levels (hyperglycemia). The disease is caused by a number of factors that either result in defects in insulin secretion or errors in insulin action, and occasionally both of these occurrences can cause hyperglycemia at the same time. Additionally, diabetic patients frequently produce advanced glycation end-products (AGEs), which can hasten the onset of osteoporosis (Yamagishi, 2011). Research on diabetes treatments is being influenced by the desire to discover inhibitors that can stop or delay carbohydrate hydrolysis with enzymes like alpha-glucosidases and lower the accumulation of sugar. Studies have shown that the majority of naturally occurring antioxidants found in plant sprouts can act as oxidative stress defence mechanisms and inhibit the primary enzymes that hydrolyze carbohydrates into simple sugars (Basha et al., 2017).
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17.6 Food Safety Issues of Allium Sprouts Sprouts provide a unique risk to food safety because infections thrive in the same warm, wet, and nutrient-rich settings required to create them. Because of the damp conditions of sprouting, decay bacteria can become a significant issue with pre- and post-germinated crops. Between 1996 and July 2016, there were about 46 outbreaks in the US that were linked to sprouts, resulting in 2474 illnesses, 187 hospitalisations, and three fatalities, including two outbreaks with the confirmed pathogen Listeria monocytogenes (Gubernot, 2015). High-quality vegetable selection, calcium hypochlorite treatment, adherence to stringent sanitation procedures during production, and proper storage of the harvested sprouts are all part of the prevention strategy. Irrigation water that has been chlorinated or ozonated may be utilised to minimise the incidences of seed rot. Also, the Institute for Food Safety and Health (IFSH), a nationally recognized leader in food safety, and the U.S. Food and Drug Administration formed the Sprout Safety Alliance (SSA) External Link Disclaimer to help sprout producers identify and implement best practices for the secure production of sprouts. The Alliance is developing a core curriculum for the group of sprout producers as well as outreach and training events. Onions and other sprouted allium species are typically unattractive to consumers and are therefore thrown away as waste. As direct disposal of onions to landfills is not recommended, decayed onions are unsuitable as organic fertilizers due to their susceptibility to phytopathogens like Sclerotium cepivorum, and they should not be used as animal feed due to their strong odour. One way to reduce environmental loading is to recover quercetin from discarded onions (Khiari et al., 2009). Additionally, sprouted and decayed onions have a high moisture content, making their removal by combustion more expensive. Due to the associated health benefits of onion waste (Corzo-Martínez et al., 2007), growers, breeders, and the food- processing industry have suggested using it as a food ingredient (Roldán et al., 2008). On the price and applicability of quercetin extraction from the waste of sprouted allium species, more research is advised.
References Abdalla, A. A., & Mann, L. K. (1963). Bulb development in the onion (Allium cepa L.) and the effect of storage temperature on bulb rest. Hilgardia, 35, 85–112. Anonymous, Annual Research Report. (2008–2009). AICRP on Post Harvest Technology. Dr. Panjabrao Deshmukh Krishi Vidyapeeth, . Arguello, J. A., Ledesma, A., & Bottini, R. (1991). Hormonal regulation of dormancy in garlic (Allium sativum L.) cv. Rosado Paraguayo. Agriscientia, 8, 9–14. Arguello, J. A., Falcon, L. R., Seisdedos, L., Milrad, S., & Bottini, R. (2001). Morphological changes in garlic (Allium sativum L.) micro-bulblets during dormancy and sprouting as related to peroxidase activity and gibberellin GA3 content. Biocell, 25, 1–9. Basha, S. C., Babu, K. R., Madhu, M., & Gopinath, C. (2017). In vitro antidiabetic activity of sulforaphane. BMC Pharmacology and Toxicology, 3, 47–49.
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Benkeblia, N. (2000). Phenylalanine ammonia-lyase, peroxidase, pyruvic acid and total phenolics variations in onion bulbs during long-term storage. LWT – Food Science & Technology, 33, 112–116. Benkeblia, N., & Shiomi, N. (2004). Chilling effect on soluble sugars, respiration rate, total phenolics, peroxidase activity and dormancy of onion bulbs. Scientia Agricola, 61, 281–285. Benkeblia, N., Varoquaux, P., Shiomi, N., & Sakai, H. (2002). Storage technology of onion bulbs cv. Rouge Amposta: effects of irradiation, maleic hydrazide and carbamate isopropyl, N-phenyl (CIP) on respiration rate and carbohydrates. International Journal of Food Science and Technology, 37, 169–175. Benkeblia, N., Ueno, K., Onodera, S., & Shiomi, N. (2005). Variation of fructo-oligosaccharides and their metabolizing enzymes in onion bulb (Allium cepa L. cv. Tenshin) during long-term storage. Journal of Food Science, 70, 208–214. Brewster, J. L. (2008). Onions and other vegetable alliums (2nd ed.). CAB International. Burba, J. L. (1993). Manual de produccion de semillas hort ´ ´ıcolas. In: Crnko, J. (Ed.), Produccion de “semilla” de ajo. INTA—EE La Consulta, Men-´ doza Centers for Disease Control, Prevention (US); National Immunization Program (Centers for Disease Control and Prevention). (2005). Epidemiology and prevention of vaccine-preventable diseases. Department of Health & Human Services, Public Health Service, Centers for Disease Control and Prevention. Chope, G. A., Terry, L. A., & White, P. J. (2007). The effect of the transition between controlled atmosphere and regular atmosphere storage on bulbs of onion cultivars SS1, Carlos and Renate. Postharvest Biology and Technology, 44, 228–239. Chung, D. M., Chung, Y. C., Maeng, P. J., & Chun, H. K. (2011). Regioselective deglycosylation of onion quercetin glucosides by Saccharomyces cerevisiae. Biotechnology Letters, 33, 778–783. Corzo-Martínez, M., Corzo, N., & Villamiel, M. (2007). Biological properties of onions and garlic. Trends in Food Science & Technology, 18, 609–625. Doss, A. (2009). Preliminary phytochemical screening of some Indian Medicinal Plants. Ancient Science of Life, 29(2),12–16. Gubb, I. R., & MacTavish, H. S. (2002). Onion pre and postharvest considerations. In H. D. Currah & L. Rabinovich (Eds.), Allium crop science: Recent advances (pp. 233–265). CAB International. Gubernot, D. Memorandum of Record: 2015–2016, Sprout and Sprout-Derived Product Related Outbreak Data, July 2016. Food and Drug Administration. Hamauzu, Y., Nosaka, T., Ito, F., Suzuki, T., Torisu, S., Hashida, M., Fukuzawa, A., Ohguchi, M., & Yamanaka, S. (2011). Physicochemical characteristics of rapidly dried onion powder and its anti-atherogenic effect on rats fed high-fat diet. Food Chemistry, 129, 810–815. Hamilton, M. J., & VanderStoep, J. (1979). Germination and nutrient composition of Alfalfa seeds. Journal of Food Science, 44, 443–445. Hanlon, P. R., & Barnes, D. M. (2011). Phytochemical composition and biological activity of 8 varieties of radish (Raphanus sativus L.) sprouts and mature taproots. Journal of Food Science, 76, 185–192. Hurst, W. C., Shewfelt, R. L., & Schuler, G. A. (1985). Shelf-life and quality changes in summer storage onions (Allium cepa). Journal of Food Science, 50, 761–763. Hussien, Y., & Gebhard, B. (2007). Dormancy and sprouting in onion Allium cepa L.) bulbs. I. Changes in carbohydrate metabolism. Journal of Horticultural Science and Biotechnology, 82, 89–96. Insani, M. E., Cavagnaro, P. F., Salomon, V. M., Langman, L., Sance, M., Pazos, A. A., & Galmarini, C. R. (2016). Variation for health-enhancing compounds and traits in onion (Allium cepa L.) germplasm. Journal of Food and Nutrition Sciences, 7, 577–591. Kevers, C., Falkowski, M., Tabart, J., Defraigne, J. O., Dommes, J., & Pincemail, J. (2007). Evolution of antioxidant capacity during storage of selected fruits and vegetable. Journal of Agricultural and Food Chemistry, 55, 8596–8603. Khiari, Z., Makris, D. P., & Kefalas, P. (2009). An investigation on the recovery of antioxidant phenolics from onion solid wastes employing water/ethanol–based solvent systems. Food and Bioprocess Technology, 2, 337–343. Majid, I., & Nanda, V. (2017). Instrumental texture and flavonoid profile of paste developed from sprouted onion varieties of Indian origin. International Journal of Food Properties, 20(11), 2511–2526.
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Manchali, S., Murthy, K. N. C., & Patil, B. S. (2012). Crucial facts about health benefits of popular cruciferous vegetables. Journal of Functional Foods, 4(1), 94–106. Mark, G. L., Gitaitis, R. D., & Lorbeer, J. W. (2002). Bacterial diseases of onion. In H. D. Rabinovich & L. Currah (Eds.), Allium crop science: Recent advances (pp. 269–270). CAB International. Mashayekhi, K., Chiane, S. M., Mianabadi, M., Ghaderifar, F., & Mousavizadeh, S. J. (2015). Change in carbohydrate and enzymes from harvest to sprouting in garlic. Food Science & Nutrition published by Wiley Periodicals, Inc, pp. 370–376. Naji, E. A., Jasna, C. B., Gordana, C., Vesna, T. S., Jelena, V., & Ilic, N. (2017). Powdered barley sprouts: Composition, functionality and polyphenol digestibility. International Journal of Food Science and Technology., 52(1), 231–238. Öpik, H., & Rolfe, S. A. (2005). The physiology of flowering plants (4th ed). Cambridge University Press, 392 pp Parvu, M., Toiu, A., Vlase, L., & Alina, P. E. (2010). Determination of some polyphenolic compounds from Allium species by HPLC-UV-MS. Natural Product Research, 24(14), 1318–2134. Qiu, D. Y., & Zhang, Z. J. (2016). Effect of isolated garlic sprouts on germination properties of Angelica sinensis seeds under simulated continuous cropping stress, p. 47. Ramos, F.A., Takaishi, Y., Shirotori, M., Kawaguchi,Y., Kawaguchi, Y., Tsuchiya, K., Shibata, H., Higuti, T., Tadokoro, T., & Takeuchi, M. (2006). Antibacterial and antioxidant activities of quercetin oxidation products from yellow Onion (Allium cepa) skin. Journal of Agricultural and Food Chemistry, 54, 3551–3557. Reed, E., Ferreira, C.M., Bell, R., Brown, E.W., & Zheng, J. (2018). Plant-Microbe and Abiotic Factors Influencing Salmonella Survival and Growth on Alfalfa Sprouts and Swiss Chard Microgreens. Applied and Environmental Microbiology, 84(9), 2814–2817. Roldán, E., Sánchez-Moreno, C., Ancos, B. D., & Cano, M. P. (2008). Characterization of onion (Allium cepa L.) by–products as food ingredients with antioxidant and anti-browning properties. Food Chemistry, 108, 907–916. Sangronis, E., & Machado, C.J. (2007). Influence of germination on the nutritional quality of Phaseolus vulgaris and Cajanus cajan. LWT - Food Science and Technology, 40(1), 116–120. Selvaraj, S. (1976). Onion: Queen of the kitchen. Kisan World, 3(12), 32–34. Sharma, K., Assefa, A.D., Kima, S., Koa, E.Y., & Parka, S.W. (2014). Change in chemical composition of onion (Allium cepa L. cv. Sunpower) during post-storage under ambient conditions. New Zealand Journal of Crop and Horticultural Science, 42:2, 87–98. Sharma, K., Assefa, A. D., Ko, E. Y., Lee, E. T., & Park, S. W. (2015). Quantitative analysis of flavonoids, sugars, phenylalanine and tryptophan in onion scales during storage under ambient conditions. Journal of Food Science and Technology, 52, 2157–2165. Swieca, M., & Gawlik-Dziki, U. (2015). Effects of sprouting and postharvest storage under cool temperature conditions on starch content and antioxidant capacity of green pea, lentil and young mung bean sprouts. Food Chemistry, 185, 99–105. Thomson, M., Al-Qattan, K. K., Divya, J. S., & Ali, M. (2016). Anti-diabetic and anti-oxidant potential of aged garlic extract (AGE) in streptozotocin-induced diabetic rats. BMC Complementary Medicine and Therapies, 16:17. Vazquez-Barrios, M. E., Lopez-Echevarria, G., MercadoSilva, E., Castano-Tostado, E., & Leon- Gonzalez, F. (2006). Study and prediction of quality changes in garlic cv. Perla (Allium sativum L.) stored at different temperatures. Scientia Horticulturae, 108, 127–132. Wang, Z. X., Dong, P. C., Sun, T. T., Xu, X. R., Ma, L., Huang, Y. M., & Lin, X. M. (2011). Comparison of lutein, zeaxanthin and b -carotene level in raw and cooked foods consumed in Beijing. Zhonghua yu fang yi xue za zhi, 45, 64–67. Wareing, P. F., & Sannders, P. F. (1971). Hormones and dormancy. Annual Review of Plant Physiology, 22, 261–288. Xihong, L., Li, L., Zhaojun, B., Xiuli, W., & Li, Z. (2010). Improved keeping quality of fresh- cut garlic sprouts by modified atmosphere packaging. In XXVIII International Horticultural Congress—IHC, pp. 77–178. Yamagishi, S. I. (2011). Role of advanced glycation end products (AGEs) in osteoporosis in diabetes. Current Drug Targets, 12, 2096–2102.
Chapter 18
Onion Sprouts Bababode Adesegun Kehinde, Oluwakemi Igiehon, Adekanye Oluwabori, and Ishrat Majid
18.1 Introduction Onions are biennial bulb plants belonging to the Allium genus and “bulby” appearance. As a plant, its bulb is naturally situated at its bottom section and its bluish- green hollow leaves are distributed in a fan-like structure. It is cultivated around the world and is also managed as a perennial or annual crop that is harvested at its first season of growth. Different species in different parts of the world have been recognized to belong to the Allium genus. They include A. vavilovii and A. asarense of Turkmenistan and Iran regions, A. canadense associated with Canada, A. fistulosum whose origin relates to Japan, and A. proliferum of Egyptian roots. However, the A. cepa is the seemingly most prominent with regards to cultivation.
B. A. Kehinde (*) Food Processing Center, University of Nebraska-Lincoln, Lincoln, NE, USA O. Igiehon Department of Microbiology and Immunology, Louisiana State University, Shreveport, LA, USA A. Oluwabori Department of Veterinary and Biomedical Sciences, Mississippi State University, Starkville, MS, USA I. Majid Department of Food Technology, Islamic University of Science and Technology, Awantipora, J&K, India © The Author(s), under exclusive license to Springer Nature Switzerland AG 2023 I. Majid et al. (eds.), Advances in Plant Sprouts, https://doi.org/10.1007/978-3-031-40916-5_18
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18.2 Diverse Applications of Onions Onions are versatile in functionalities, making them useful for food and non-food uses. Their volatile oils have been studied to be linked to these. Onions have found applications in diverse fields of agronomy, cosmetics, pharmaceutics, and medical sciences. Khater et al. (2013) used onion oil as an insecticide to inhibit the second and third phases of larval development of oestrid fly, Cephalopina titillator (Clark). The study reported mortality rates of 86 and 78% on the tested 50 larvae after 48 h. Mansour and Abdel-Hamid (2015) compared oil extracts from different plant materials such as Cumin seeds (Cuminum cyminum Linn.), Parley seeds (Petroselinum sativum L.), Sweet Basil (Ocimum basilicum Linn.), Marjoram (Origanum vulgare L.), Geranim (Pelargonium radula Cav), Chamomile Blue whole plant (Matricaria chamomilla L.), and Onion leaves (Allium cepa L) against 3rd nymphal holometabolous of Schistocerca gregaria (Forskål) (Orthoptera: Acrididae), commonly referred to as desert locust. The oils were applied using the bait application technique and their relative percentage potency compared with Methomyl were reported to be 100, 93.6, 90.7, 80.5, 34.3, 29.3, and 28.8 for Cumin, Sweet Basil, Onion, Geranium, Marjoram, Chamomile, and Parsley. Only oil extracts from Cumin and Sweet Basil were found to be more potent than the one obtained from Onion. Gharsan et al., 2018 examined the toxicity of oil extracts from five different plant materials against grain pests namely, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) and Oryzaephilus surinamensis (L.) (Coleoptera: Silvanidae). Oils from brown galingale (Cyperusfuscus, or saad L, Cyperaceae), caraway (Carum carvi [Lindl.] H. Wolff, Apiaceae), flax (Linum usitatissimum Mill., Linaceae), onion (Allium cepa L., Amaryllidaceae) and lavender (Lavandula angustifolia Mill., Lamiaceae) were used. At a concentration of 4 μL/mL, mortality percentage results against Oryzaephilus surinamensis (L.) were 100 ± 0.00 for caraway and onion, while flaxseed, lavender and saad were 95 ± 2.88, 90 ± 0.00, and 90 ± 0.00 respectively. Against Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), mortality percentage results of 70 ± 0.00, 90 ± 5.77, 55 ± 2.88, 95 ± 2.88, and 60 ± 11.54 were reported. Draelos (2008) examined how the gel obtained from extract revamps the outlook of post-surgical scars in human subjects with symmetrical seborrheic keratoses. The gel extracts were used on the patients at their surgical sites having a minimum of 8 mm-diameter of symmetrical seborrheic keratoses on the left and right upper chest with subsequent examinations after 2 to 3 weeks of healing. The study reported a significant improvement in the global appearance, texture, redness, and softness of the scars by the onion gel extract. Cho et al. (2010) examined the in vivo and in vitro impacts of onion extract on skin healing. The study involved the application of quercetin and onion extract on fibroblast proliferation, and expression of type I collagen and matrix metalloproteinase-1 (MMP-1). The study reported that quercetin and onion extract decreased fibroblast proliferation rates through a dose-dependent fashion, and though the MMP-1 expression was markedly enhanced by both components in vivo and in vitro, significant alterations were not exerted on type I collagen.
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Pikuła et al. (2014) analyzed how onion extract and enoxaparin affects β1 integrin expression, proliferation, and apoptosis on human fibroblasts. The human Fibroblast cell lines (46 BR.1 N) were dosed with varying amounts of onion [Allium cepaL. (Alliaceae)] extract (50, 250, 1000 mg/mL) and/or enoxaparin sodium (500, 100, and 20 mg/mL) and the β1 integrin expression and apoptosis evaluated by flow cytometry while [3H]-thymidine incorporation assay was used to analyze cell proliferation. The study reported that both treatments inhibited human fibroblast proliferation and that at an amount of 250 mg/mL, the extract obtained from onion showed a strong inhibition to cell proliferation by a 50.8% reduction. The study concluded that onion extracts are potentially useful for the management of keloids and scars that are hypertrophic in nature.
18.3 Onion Nutraceuticals and Biological Benefits Onions contain a wide variety of biologically active components such as organosulfur, phenolics, saponins, and polysaccharides known to possess antidiabetic, antimicrobial, anti-obesity, anti-inflammatory, and antioxidant activities and serve to protect different body systems such as respiratory, cardiovascular, digestive, and reproductive systems (Kehinde & Sharma, 2018; Zhao et al., 2021). In addition, several anthocyanins, alk(en)yl cysteine sulphoxides, and flvaonols such as cyanidins, (propen-1-yl)-L-cysteine sulphoxide, propyl-L-cysteine sulphoxide, methyl- L- cysteine sulphoxide, isorhamnetin, kaempferol and peonidin (Griffiths et al., 2002).
18.4 Antidiabetic Effects Lee et al. (2013) examined how onion juice affected serum glucose levels of in vivo using rats administered with streptozotocin by intraperitoneal injection to raise their blood glucose levels to ≥300 mg/dL. The Sprague-Dawley rats showed glucose blood levels of 366 ± 5 mg/dL at the first day of experimentation and 315 ± 8 mg/ dL (14% reduction) at the sixth day with orally administered onion juice at 15 mL/ kg b.w. Jung et al. (2011) studied how onion peel extract affects insulin resistance and hyperglycemia in diabetic Sprague-Dawley rats. The rats were orally administered with high AIN-93G fat régime (bearing about 41.2% lipid) and subsequently dosed with 40 mg/kg b.w streptozotocin by intraperitoneal injection. The onion peel extracts were obtained using 60% on onion bulbs at 50 °C and a pH of 5.5 for a duration of 3 hours and fed at 1% concentration to the laboratory animals which had above 126 mg/dL of fasting blood glucose levels. The study reported an insulin sensitizing effect of the extracts by increased levels of glycogen in the skeletal muscle and liver from quantitative Real-Time Polymerase Chain Reaction (RT-PCR)
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analysis which indicated enhanced GLUT4 and insulin receptor expression in muscle tissues. In addition, hepatic protein expressions, plasma free fatty acids, and oxidative stress as evaluated by malondialdehyde formation and superoxide dismutase activity were all found to be significantly reduced by the onion extract administration. The study also concluded that the extract might have enhanced insulin resistance and glucose response lined with diabetes by inhibiting oxidative stress, assuaging metabolic dysregulation of free fatty acids, up-regulating glucose uptake at peripheral tissues and/or down-regulating inflammatory gene expression in liver.
18.5 Antihypertensive Effects Quercetin and several sulfur-bearing compounds such as S-methylcysteine sulfoxide, S-propylcysteine sulfoxide and S-propenylcysteine sulfoxide along with other alkyl cysteine sulfoxides have been reported to be of possible advantages for management of hypertension and associated disorders. Sakai et al. (2003) investigated how onion affected the status of stroke-prone and L-NAME (NG-nitro-L-arginine methyl ester) induced-hypertensive rats. Sprague-Dawley male rats, about 6 weeks old, were induced by oral administration with NG-nitro-L-arginine methyl ester at a concentration of 50 mg/kg BW/day after which they were fed with 5% dried onion in their diets. The onion was found to improve the nitrite/nitrate derivatives of nitric oxide excreted in the urine of the rats. However, the antihypertensive action of the onion was speculated to have possibly resulted from antioxidative action of the dried onion in L-NAME initiated-hypertensive animals. Olayeriju et al. (2015) examined how ethyl acetate extracts of red onion affected the biochemical and hemodynamic properties of normotensive albino rats. The extracts were orally administered at concentrations of 10-, 20-, or 40 mg/kg for 14 days and the experimentation showed a dose-dependent relationship between the onion extracts and their anti-hypertensive effects. The pulse pressure, mean arterial blood pressure, heart rate, diastolic blood pressure and systolic blood pressure of the examined test animals were found to have a dose-dependent decrease.
18.6 Antioxidant Benefits Several classes of nutraceuticals such as oleoresins, polysaccharides, flavonoids, sulfides, and nitrogenous compounds in onions have been studied to be of potential antioxidative effects (Rafiq et al., 2022). Such compounds have been observed to scavenge free radicals and reverse oxidation and associated disorders in animals. Zhou et al. (2022) purified polysaccharides from onion through multiple unit operations such as deproteinization, alcohol precipitation, hot water extraction, homogenization, and drying and examined in vitro the potentials of the extract and its
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derivatives to inhibit oxidation. Tests such as reduction ability, anti-lipid peroxidation, and the superoxide anion and hydroxyl radical scavenging strength proved positive regarding the antioxidant profile. Chernukha et al. (2021) examined the in vivo antioxidant effect of ethanolic extracts obtained from yellow onion husk. The study involved 20 male Wistar albino rats that were arbitrarily sectioned in two equal classes viz. an experimental group that were orally administered the extract and a control group. After a 188-day period, the experimental group was reported to have their brain and liver antioxidant system affected as shown by elevations in superoxide dismutase and catalase activities by 79.1 and 300% for the brain, and 79.1 and 44.4% for the liver respectively.
18.7 Onion Antimicrobials The opening or cutting of onion tissues initiates enzymes such as S-alk(en)yl cysteine sulfoxides lyase or alliinase to hydrolyze S-alk(en)yl-l-cysteine sulfoxides with an eventual synthesis of pyruvic acid, ammonia, and sulfenic acids which in turn spontaneously react to produce thiosulfinates by condensation (Loredana et al., 2019). These unstable compounds have been attributed to the antimicrobial properties of onions against pathogens within and without food systems. Faluyi et al. (2020) studied the preservative effects of diced onion on unrefrigerated chicken meat. The study involved lacing an experimental group of 150 g of freshly diced red onions for each kilogram of meat and the control group without any onions with a periodic evaluation of their microbial profiles every 12 hours for 36 hours. The study reported that during the storage duration, the onion-laced had markedly reduced counts relative to the control for the 12th and 24th hour, though no significant differences in their microbial counts were found at the end of the storage study. However, microbes such as Klebsiella spp., Proteus vulgaris, Enterobacter aerogenes, and Escherichia coli. had lower percentage prevalence during storage for the onion-laced samples. Loredana et al. (2019) examined the antibacterial effects of Vatolla, Alife, and Montoro onion varieties native to Italy using the halo test on agar plates. Bacteria such as Escherichia coli (DSM 8579), Pseudomonas aeruginosa (DSM 50071), Listeria innocua (DSM 20649), Staphylococcus aureus (DSM 25923), and Bacillus cereus (DSM 43984 and 4313). The ethanolic extracts were reported to be actionable in inhibiting Gram-negative bacteria (P. aeruginosa) and Gram-positive species employed in the order of Staphylococcus aureus