Roselle (Hibiscus Sabdariffa): Chemistry, Production, Products, and Utilization 9780128221006, 0128221003

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Roselle (Hibiscus sabdariffa)

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Roselle (Hibiscus sabdariffa) Chemistry, Production, Products, and Utilization

Edited by

Abdalbasit Adam Mariod Indigenous Knowledge and Heritage Center, Ghibaish College of Science & Technology, Ghibaish, Sudan; College of Sciences and Arts-Alkamil, University of Jeddah, Alkamil, Saudi Arabia

Haroon Elrasheid Tahir School of Agricultural Equipment Engineering, School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China

Gustav Komla Mahunu Department of Food Science and Technology, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana

Academic Press is an imprint of Elsevier 125 London Wall, London EC2Y 5AS, United Kingdom 525 B Street, Suite 1650, San Diego, CA 92101, United States 50 Hampshire Street, 5th Floor, Cambridge, MA 02139, United States The Boulevard, Langford Lane, Kidlington, Oxford OX5 1GB, United Kingdom Copyright © 2021 Elsevier Inc. All rights reserved. No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, recording, or any information storage and retrieval system, without permission in writing from the publisher. Details on how to seek permission, further information about the Publisher’s permissions policies and our arrangements with organizations such as the Copyright Clearance Center and the Copyright Licensing Agency, can be found at our website: www.elsevier.com/permissions. This book and the individual contributions contained in it are protected under copyright by the Publisher (other than as may be noted herein). Notices Knowledge and best practice in this field are constantly changing. As new research and experience broaden our understanding, changes in research methods, professional practices, or medical treatment may become necessary. Practitioners and researchers must always rely on their own experience and knowledge in evaluating and using any information, methods, compounds, or experiments described herein. In using such information or methods they should be mindful of their own safety and the safety of others, including parties for whom they have a professional responsibility. To the fullest extent of the law, neither the Publisher nor the authors, contributors, or editors, assume any liability for any injury and/or damage to persons or property as a matter of products liability, negligence or otherwise, or from any use or operation of any methods, products, instructions, or ideas contained in the material herein. British Library Cataloguing-in-Publication Data A catalogue record for this book is available from the British Library Library of Congress Cataloging-in-Publication Data A catalog record for this book is available from the Library of Congress ISBN: 978-0-12-822100-6 For Information on all Academic Press publications visit our website at https://www.elsevier.com/books-and-journals

Publisher: Nikki Levy Acquisitions Editor: Megan Ball Editorial Project Manager: Mona Zahir Production Project Manager: Kumar Anbazhagan Cover Designer: Christian J. Bilbow Typeset by MPS Limited, Chennai, India

Contents List of contributors ................................................................................................ xiii

CHAPTER 1 Breeding, genetic diversity, and safe production of Hibiscus sabdariffa under climate change ............................................................. 1 1.1 1.2

1.3 1.4 1.5

Gustav Komla Mahunu Abbreviation .................................................................................. 1 Introduction ....................................................................................1 Breeding and genetic diversity ......................................................3 1.2.1 Breeding .............................................................................. 3 1.2.2 Genetics diversity................................................................ 4 Safe production of roselle in climate change ................................5 Potential sources of heavy metals and microbial contamination of roselle.................................................................8 Conclusion ......................................................................................9 Acknowledgment ......................................................................... 10 Conflict of interest....................................................................... 10 References.................................................................................... 10

CHAPTER 2 Harvesting, storage, postharvest management, and marketing of Hibiscus sabdariffa........................ 15 2.1 2.2

2.3

2.4

Mildred Osei-Kwarteng, Joseph Patrick Gweyi-Onyango and Gustav Komla Mahunu Introduction ..................................................................................16 Harvesting of roselle ....................................................................17 2.2.1 Determinants of harvest maturity of roselle..................... 17 2.2.2 Harvesting of leaves and tender shoots ............................ 18 2.2.3 Harvesting of calyces........................................................ 19 2.2.4 Harvesting of seeds ........................................................... 22 2.2.5 Harvesting stems for fiber ................................................ 22 Postharvest management of harvested roselle .............................23 2.3.1 Postharvest management of leaves and tender shoots ..... 23 2.3.2 Postharvest management of calyces ................................. 23 2.3.3 Postharvest management of seeds .................................... 25 2.3.4 Postharvest management of stem (fiber) .......................... 26 Storage of useful parts of roselle .................................................27 2.4.1 Storage of leaves and tender shoots after harvest ............ 27

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2.4.2 Storage of harvested calyces............................................. 27 2.4.3 Storage of harvested seeds................................................ 27 2.5 Marketing of roselle produce and products .................................28 2.5.1 Marketing of leaves and tender shoots, calyces, seeds, and fiber ................................................................. 28 2.6 Conclusion ....................................................................................30 References.................................................................................... 30

CHAPTER 3 Effect of pests and diseases on Hibiscus sabdariffa quality ........................................................ 33 3.1 3.2

3.3

3.4

Gustav Komla Mahunu, Maurice Tibiru Apaliya and Mildred Osei-Kwarteng Introduction ..................................................................................33 Pests of roselle..............................................................................34 3.2.1 Types of pests ................................................................... 34 3.2.2 Effect of pests on roselle .................................................. 35 3.2.3 Prevention and control of pests ........................................ 38 Diseases of roselle........................................................................40 3.3.1 Types of diseases .............................................................. 40 3.3.2 Effect of diseases on roselle ............................................. 41 3.3.3 Prevention and control of diseases ................................... 42 Conclusion ....................................................................................43 References.................................................................................... 43

CHAPTER 4 Measurement and maintenance of Hibiscus sabdariffa quality ........................................................ 47 Joseph Patrick Gweyi-Onyango, Mildred Osei-Kwarteng and Gustav Komla Mahunu 4.1 Background...................................................................................47 4.2 Quality and quality maintenance of the Hibiscus sabdariffa......49 4.2.1 Anthocyanins as an essential component of Hibiscus sabdariffa quality ............................................... 50 4.2.2 Microbial and antimicrobial activities and Hibiscus sabdariffa quality .............................................................. 51 4.2.3 Influence of plant growth, age, and harvesting on Hibiscus sabdariffa quality ............................................... 52 4.2.4 Antinutritional components and the prevailing factors on Hibiscus sabdarrifa quality ............................. 54 4.3 Maintenance of quality of secondary metabolites of Hibiscus sabdariffa.......................................................................55

Contents

4.4 Methods of measurements of bioactive compounds in Hibiscus sabdariffa.......................................................................57 References.................................................................................... 59

CHAPTER 5 Composition of Hibiscus sabdariffa calyx, pigments, vitamins...................................................... 69 Abdalbasit Adam Mariod, Haroon Elrasheid Tahir and Gustav Komla Mahunu 5.1 Introduction ..................................................................................69 5.1.1 Plant description................................................................ 69 5.1.2 The chemical composition of Hibiscus sabdariffa calyx................................................................. 70 5.1.3 Composition of Hibiscus pigments................................... 71 5.1.4 Composition of Hibiscus sabdariffa vitamins.................. 72 5.1.5 Parts used medically ......................................................... 72 5.1.6 Pharmacological and therapeutic properties of hibiscus ......................................................................... 73 References.................................................................................... 74

CHAPTER 6 Hibiscus sabdariffa: protein products, processing, and utilization......................................... 77

6.1 6.2

6.3

6.4 6.5

Mildred Osei-Kwarteng, Joseph Patrick Gweyi-Onyango, Gustav Komla Mahunu, Haroon Elrasheid Tahir and Maurice Tibiru Apaliya Introduction ..................................................................................77 Protein products............................................................................79 6.2.1 Seeds.................................................................................. 79 6.2.2 Leaves................................................................................ 79 6.2.3 Calyces and general flowers ............................................. 80 Processing of protein products.....................................................80 6.3.1 Types of processing methods ........................................... 81 6.3.2 Quality of proteins from different processing methods .......... 82 Utilization of roselle protein products .........................................84 Conclusion ....................................................................................85 References.................................................................................... 85

CHAPTER 7 Volatile compounds and phytochemicals of Hibiscus sabdariffa..................................................... 91 Abubakr Musa, Haroon Elrasheid Tahir, Mohammed Abdalbasit A. Gasmalla, Gustav Komla Mahunu and Abdalbasit Adam Mariod 7.1 Introduction ..................................................................................91

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7.2 Bioactive constituents ..................................................................92 7.3 Organic acids................................................................................92 7.3.1 Ascorbic acid..................................................................... 96 7.3.2 Hydroxycitric acid ............................................................ 97 7.4 Anthocyanins................................................................................98 7.5 Flavonoids ....................................................................................99 7.6 Volatile compounds......................................................................99 7.7 Conclusion ..................................................................................107 References.................................................................................. 108

CHAPTER 8 Oil recovery from Hibiscus sabdariffa seeds .......... 113 8.1 8.2 8.3 8.4 8.5 8.6

Abdalbasit Adam Mariod, Haroon Elrasheid Tahir and Gustav Komla Mahunu Introduction ................................................................................113 Roselle components as food source...........................................114 Protein of roselle seeds ..............................................................115 Oil of roselle seeds.....................................................................115 Roselle oil seed products ...........................................................120 Conclusion ..................................................................................121 References.................................................................................. 121

CHAPTER 9 Food use of whole and extracts of Hibiscus sabdariffa................................................... 123

9.1 9.2

9.3 9.4

Gustav Komla Mahunu, Haroon Elrasheid Tahir, Mildred Osei-Kwarteng, Abdalbasit Adam Mariod and Joseph Patrick Gweyi-Onyango Abbreviations............................................................................. 123 Introduction ................................................................................124 Use of whole roselle plant constituents in food ........................124 9.2.1 Calyx ............................................................................... 124 9.2.2 Leaves.............................................................................. 129 9.2.3 Roselle seeds ................................................................... 129 Other functional food applications roselle extracts...................130 Conclusion ..................................................................................131 References.................................................................................. 132

CHAPTER 10 Nutritional properties and feeding values of Hibiscus sabdariffa and their products ................... 137 Maurice Tibiru Apaliya, Emmanuel Kwaw, Gustav Komla Mahunu, Mildred Osei-Kwarteng, Richard Osae and Michael Azirigo 10.1 Introduction ................................................................................137

Contents

10.2 Nutritional properties of Hibiscus sabdariffa ............................138 10.2.1 Nutritional composition ................................................ 138 10.2.2 Organic acids................................................................. 142 10.3 Feeding values of Hibiscus sabdariffa ......................................145 10.3.1 Health benefits .............................................................. 145 10.3.2 Uses and value .............................................................. 147 10.4 Products of Hibiscus sabdariffa.................................................148 References.................................................................................. 150

CHAPTER 11 Medicinal and therapeutic potential of roselle (Hibiscus sabdariffa) ................................................ 155

11.1 11.2 11.3 11.4

11.5 11.6

Muhammad Arslan, Muhammad Zareef, Haroon Elrasheid Tahir, Allah Rakha, Zou Xiaobo and Gustav Komla Mahunu Introduction ................................................................................156 Roselle bioactive compounds and their therapeutic benefits .......................................................................................157 Roselle uses in traditional medicine ..........................................158 Medicinal and therapeutic health benefits of roselle ................159 11.4.1 Antihypertensive activity ............................................ 159 11.4.2 Anti-inflammatory activity ......................................... 159 11.4.3 Antiobesity activity ..................................................... 161 11.4.4 Antidiabetic activity.................................................... 162 11.4.5 Nephroprotective activity............................................ 164 11.4.6 Hepatoprotective activity ............................................ 165 11.4.7 Cardioprotective activity............................................. 167 11.4.8 Renal effects, uricosuric effect, and hyperuricemia.............................................................. 169 11.4.9 Treatment of anemia ................................................... 170 11.4.10 Cancer preventive activity .......................................... 171 11.4.11 Uses against cadmium poisoning ............................... 172 Future perspectives.....................................................................174 Conclusion ..................................................................................175 References.................................................................................. 175

CHAPTER 12 Hibiscus sabdariffa interactions and toxicity.......... 187 Haroon Elrasheid Tahir, Gustav Komla Mahunu, Zou Xiaobo and Abdalbasit Adam Mariod 12.1 Introduction ................................................................................187 12.2 Hibiscus
drug interactions........................................................189

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12.3 12.4 12.5 12.6

Hibiscus sabdariffa toxicology ..................................................191 Effect of hibiscus on kidney and liver functions ......................193 Other adverse effects..................................................................194 Conclusion ..................................................................................195 References.................................................................................. 195

CHAPTER 13 Conventional and rapid methods for measurement of total bioactive components and antioxidant activity in Hibiscus sabdariffa.............. 199 13.1 13.2

13.3 13.4 13.5 13.6

Huang Xiaowei, Li Zhihua, Haroon Elrasheid Tahir, Zou Xiaobo, Shi Jiyong, Xu Yiwei and Zhai Xiaodong Introduction ................................................................................199 Spectrophotometric techniques ..................................................201 13.2.1 Conventional measurement of total phenolic content ........................................................................... 201 13.2.2 Measurement of total flavonoid content and total anthocyanin content ...................................................... 204 13.2.3 Measurement of antioxidant activities.......................... 204 Near-infrared spectroscopy ........................................................205 Chemometrics analysis...............................................................206 Determination of phenols, flavonoid, anthocyanins, and antioxidant activities in roselle ..................................................207 Conclusion ..................................................................................209 References.................................................................................. 210

CHAPTER 14 Hibiscus sabdariffa extract: antimicrobial prospects in food pathogens and mycotoxins management .... 215

14.1 14.2 14.3 14.4 14.5 14.6 14.7 14.8

Lydia Quansah, Gustav Komla Mahunu, Haroon Elrasheid Tahir, Maurice Tibiru Apaliya, Mildred Osei-Kwarteng and Abdalbasit Adam Mariod Introduction ................................................................................216 Antioxidant properties in H. sabdariffa.....................................216 Antimicrobial potentials of H. sabdariffa..................................218 Mycotoxin production and economic losses to food.................220 Mycotoxin effect on human health............................................220 Synthetic fungicides: implications on food quality and consumer health..........................................................................222 H. sabdariffa extract: biocontrol agent against pathogens and mycotoxins ..........................................................................222 Conclusion ..................................................................................223 References.................................................................................. 223

Contents

CHAPTER 15 Ethnobotanical uses, fermentation studies and indigenous preferences of Hibiscus sabdariffa....... 231 15.1 15.2 15.3

15.4

15.5

Haroon Elrasheid Tahir, Gustav Komla Mahunu, Zou Xiaobo and Abdalbasit Adam Mariod Introduction ................................................................................231 Ethnobotanical uses....................................................................234 Fermented studies.......................................................................234 15.3.1 Seeds fermented products ............................................. 234 15.3.2 Roselle calyx’s fermented products.............................. 240 15.3.3 Uses of roselle in fermented milk ................................ 243 15.3.4 Uses of roselle in baked products................................. 244 Indigenous preferences...............................................................246 15.4.1 Roselle beverage ........................................................... 246 15.4.2 Seeds.............................................................................. 247 15.4.3 Leaves............................................................................ 248 15.4.4 Fermented milk ............................................................. 248 Conclusion ..................................................................................248 References.................................................................................. 249

Index ......................................................................................................................255

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List of contributors Maurice Tibiru Apaliya Department of Food Science and Postharvest Technology, Faculty of Applied Sciences, Cape Coast Technical University, Cape Coast, Ghana; Department of Hotel Catering and Institutional Management, Cape Coast Technical University, Cape Coast, Ghana Muhammad Arslan School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China Michael Azirigo Department of Agriculture for Social Change, Regentropfen College of Applied Sciences, Bongo, Ghana Mohammed Abdalbasit A. Gasmalla Department of Nutrition and Food Technology, Omdurman Islamic University, Omdurman, Sudan Joseph Patrick Gweyi-Onyango Department of Agricultural Science and Technology, Kenyatta University, Nairobi, Kenya Shi Jiyong School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China Emmanuel Kwaw Department of Food Science and Postharvest Technology, Faculty of Applied Sciences, Cape Coast Technical University, Cape Coast, Ghana Gustav Komla Mahunu Department of Food Science and Technology, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana Abdalbasit Adam Mariod Indigenous Knowledge and Heritage Center, Ghibaish College of Science & Technology, Ghibaish, Sudan; College of Sciences and Arts-Alkamil, University of Jeddah, Alkamil, Saudi Arabia Abubakr Musa Sugar Institute, University of Gezira, Wad Madani, Sudan Richard Osae Department of Food Science and Postharvest Technology, Faculty of Applied Sciences, Cape Coast Technical University, Cape Coast, Ghana Mildred Osei-Kwarteng Department of Horticulture, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana

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Lydia Quansah Department of Biotechnology, Faculty of Biosciences, University for Development Studies, Tamale, Ghana Allah Rakha National Institute of Food Science and Technology, University of Agriculture, Faisalabad, Pakistan Haroon Elrasheid Tahir School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China Zou Xiaobo School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China Zhai Xiaodong School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China Huang Xiaowei School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China; School of Agricultural Equipment Engineering, Jiangsu University, Zhenjiang, P.R. China Xu Yiwei School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China Muhammad Zareef School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China Li Zhihua School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China

CHAPTER

Breeding, genetic diversity, and safe production of Hibiscus sabdariffa under climate change

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Gustav Komla Mahunu Department of Food Science and Technology, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana

Chapter Outline Abbreviation ............................................................................................................ 1 1.1 Introduction ..................................................................................................... 1 1.2 Breeding and genetic diversity .......................................................................... 3 1.2.1 Breeding ........................................................................................3 1.2.2 Genetics diversity ............................................................................4 1.3 Safe production of Roselle in climate change .................................................... 5 1.4 Potential sources of heavy metals and microbial contamination of Roselle .......... 8 1.5 Conclusion ....................................................................................................... 9 Acknowledgment ................................................................................................... 10 Conflict of interest ................................................................................................. 10 References ............................................................................................................ 10

Abbreviation AIVs

African indigenous vegetables

1.1 Introduction African indigenous vegetables (AIVs) remain nearly important crops that support diet, health improvement, and commercial benefits to the people of Africa. These vegetables are generally considered as underutilized crop species. By description, these crops are native and adapted fruits and vegetables gathered from wild populations, relatively easy to cultivate within a short vegetation period aided with lower inputs than other exotic (temperate) vegetables. These crops are very much Roselle (Hibiscus sabdariffa). DOI: https://doi.org/10.1016/B978-0-12-822100-6.00005-7 © 2021 Elsevier Inc. All rights reserved.

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acclimatized to native environmental conditions and mainly provide livelihood support toward income, health, and nutrition. AIVs continue to be part of the solution to averting hunger, malnutrition, and impacts of climate change (Sto¨ber et al., 2017). The preference for AIVs varies across countries, thus defined by the eating habits of the peoples, availability, and quantity required for consumption (Dansi et al., 2008). Notably, approximately 1000 species of natural plant species are used as vegetables, of which the majority (80%) are leafy vegetables, with the other 20% made up of vegetables from fruits, seeds, roots and tubers, stems, and flowers (Shackleton, Pasquini, & Drescher, 2009). The quest for local innovations and discoveries of new vegetable species has significantly advanced as people search for ways to manage food insecurity and enrich their awareness of indigenous species (Maundu, Achigan-Dako, & Morimoto, 2009). Much important, the search or selection of these novel categories of AIVs has been through the introduction of species and/or depending on the native African species. Among the AIVs, Roselle in recent years has found place as a very important multipurpose indigenous vegetable across Africa. In times past, Roselle was a minor vegetable crop, but it is fast gaining attention for the food and manufacturing industries, as will be discussed subsequently. Hibiscus sabdariffa plant produces edible calyx, belonging to a large family Malvaceae (Sharma et al., 2016). It is annual but can be cultivated as a perennial plant in the tropical and subtropical areas worldwide. In addition to the bast fiber and paper pulp or calyx, it also produces leaves and seeds (Osman et al., 2011). According to Satyanarayana, Visalakshmi, Mukherjee, Priya, and Sarkar (2015), the Roselle is considered as an important crop among the bast fiber group India, holding the second position among fiber crops after jute. Generally, the fiber is mainly used for making industrial products such as sacs, twines, and carpets. Despite its quality for fiber products in Asia, Roselle is underutilized and its fiber product is less used in the Sub-Saharan Africa (SSA) (Tetteh, Ankrah, Coffie, & Niagiah, 2019). In recent times, various products from Roselle are being explored and promoted in SSA. Salt-resistant trait in Roselle fiber makes it a perfect material for cordage production (Singh, 2017), as packaging sacks, assorted paper material, upholstery, and fabric shoes and bags production (Managooli, 2009). Roselle, in recent years, has been identified as biocomposite for the manufacturing of vehicle parts and materials for construction including fiber board (Alves et al., 2010). Actually, Roselle is estimated to be 20% of bast fiber crops. In northern Ghana, Roselle is mostly known as “sobolo” or “suure.” The plant has more than 25 names in the tribal folks, which is a proof of Roselle domestication in the northern part of Ghana (Ankrah, Tetteh, Coffie, & Niagiah, 2018). Other common names like “biito” in Nankana and Frafra, “vio” among the Grushi and Kasem, and “tingyanbam” in Konkomba. Common names for Roselle vary on the basis of the geographical origin; they are Roselle, razelle, sorrel, red sorrel, Guinea sorrel, Jamaican sorrel, Indian sorrel, sour-sour, Queensland jelly plant, karkade´, Pusa hemp, rohzelu, laalambaar, sabdriqa, jelly okra, lemon bush,

1.2 Breeding and genetic diversity

and Florida cranberry (Kays, 2011; Mahadevan & Kamboj, 2009; Mohamed, Sulaiman, & Dahab, 2012; Small, 2006). It appears that East Indies is probably the origin of Roselle species (Duke, 1993), with tropical Africa having extreme diversity (Wilson & Menzel, 1964).

1.2 Breeding and genetic diversity 1.2.1 Breeding Roselle is most bred for its fiber yield (Rajasekharan, 2004). According to Wilson and Menzel (1964), Roselle is a tetraploid (2n 5 4x 5 72), the chromosomes are related to the diploid (2n 5 2x 5 36) Hibiscus cannabinus. There are two types of the Roselle plant: the H. sabdariffa var. sabdariffa (HSS) cultivated mainly for the fleshy, shiny-red caly and the H. sabdariffa var. altissima (HSA) cultivated mainly for its phloem fiber (Purseglove, 1968). There is little attention on Roselle, and information on the genetics, breeding, and production related to climate change adaptation is scarce. The world’s largest collection of Roselle accessions (628) is by the ICARCentral Research Institute for Jute and Allied Fibres, India (Mahapatra, 2008). There are two categories of the plant according to the growth habit and usage of the end products (Sharma et al., 2016). However, HSA category consists of plants with upright growth habit with branching at the lower part supported by an extended stem that has bast fiber of commercial use. On the other hand, the HSS category is described as bushy and has hefty branches, which bear fleshy calyces (Mahadevan & Kamboj, 2009). According to the morphological structure of the calyx, the HSS has been defined in various races. These characterizations are bhagalpuriensi (the calyces produced are bears green, red-streaked but inedible), intermedius and albus (calyces produced are edible with yellow green, some form of fiber also produced), and ruber (edible calyces produced are red) (Da-CostaRocha, Bonnlaender, Sievers, Pischel, & Heinrich, 2014). Before 2009, researchers in India developed eight varieties of Roselle with high fiber content based on the selection and hybridization of indigenous landraces (Kar et al., 2010). The detection of unique germplasm with extensive diversity is essential for maintaining or improving the fiber and calyx yield in the species. For that reason, sufficient characterization of available germplasm offers the most significant opportunity in crop improvement programs. Usually, morphological attribute (based on phenotypic qualities) to select germplasm is adopted across the gene bank because it has provision for easy and quick scoring (Ganopoulos, Kazantzis, Chatzicharisis, Karayiannis, & Tsaftaris, 2011). Environmental conditions often have a significant influence on the phenotypic traits; this can lead to an overestimated mixture of assessing agronomically important variables resulting from high gene 3 environment interactions (Marinoni, Akkak, Bounous, Edwards, & Botta, 2003). In the Roselle, several morphological

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qualities are known as a germplasm descriptor (Mahajan, Sapra, Umesh, Singh, & Sharma, 2000), which are much determined by the environment with limited success for genetic characterization of the species. Therefore agromorphological behaviors determined by molecular markers is a tool for better determination and more specific description. For decades now, molecular markers especially simple sequence repeats have shown considerable success in some crop species including rice (Nachimuthu et al., 2015), wheat (Chen, Min, Yasir, & Hu, 2012), pigeon pea (Kumari, Mishra, & Srivastava, 2014), peanut (Ren et al., 2014), and jute (Banerjee et al., 2012) for germplasm characterization, range of genetic evaluation and analysis of the population structure. These markers have shown many advantages above conservative morphological indicators. The advantages describe the fact that they are abundant, reproducible, environment and crop stage independent, high polymorphism, hypervariability, and codominant in nature (Sharma et al., 2016).

1.2.2 Genetics diversity In recent years, it has been reported that the dearth of variability in exotic Roselle germplasm hinders the contributions to improving genetic characters (Mohammed, Islam, Jahan, Yaakob, & Osman, 2014). The study of genetic diversity in Roselle makes data available in the increased cultivar improvement and their conservation amid the changing climate. Genetic diversity is an asset to germplasm collection; thus its estimation may depend on three approaches: morphological, biochemical, or molecular evaluation (Bhandari, Bhanu, Srivastava, Singh, & Shreya, 2017). Morphological evaluation will offer less cost and the easy assessment of measurements makes it attractive to breeders to use it for genetic improvement programs. But then, morphological evaluation proves to be labor-intensive, requires large size of plant population, exhibits a low rate of polymorphism, and is inhibited by its sensitivity to the environment with higher risks of biased estimates (Botha & Venter, 2000). As mentioned earlier, the morphological approach generates sufficient information on crop physiognomies and presents the origins of beneficial genotypes for improving the traits, regardless of the weaknesses (Camussi, Ottaviano, Calinski, & Kaczmarek, 1985). Studies on genetic diversity of Roselle (var. altissima) are rather lacking and limited with few recent reports. Some of the works include 36 accessions of wild Roselle identified in Ghana by Ankrah et al. (2018), Roselle bast fiber characterization in Kenya by Mwasiagi et al. (2014), and comparison studies of variability between kenaf and Roselle by Coffie (2017). Generally, the results of these studies showed variations in fruit morphology of Roselle across the globe (Sharma et al., 2016; Tetteh et al., 2019). The results of Tetteh et al. (2019) showed genetic variability among the plant component (seed, calyx harvests, leaves, and other yield parameters) of Roselle. Fundamentally, the special morphological features of Roselle make it able to survive under diverse growth conditions. The special morphological features include deep taproot, which enables deep soil penetration for the search of water and

1.3 Safe production of Roselle in climate change

minerals for growth. The height can reach up to 3.5 m with smooth or nearly smooth, cylinder-shaped stem, with dark green to red color characteristics. Comparatively, the leaves are green with reddish veins as well as long or short petioles, and they alternate with length between7.5 and 12.5 cm. The leaves of young seedlings and upper leaves of older plants look simple. The lower leaves are between three and five or even seven-lobed with saw-like margins. Flowers are about 12.5 cm wide with one each in the leaf axils. The flowers are yellow or buff colored with a rose or maroon eye and turn pink at senescence. The natural redcolored calyx has five large sepals with a collar (epicalyx) of between 8 and 12 slim, pointed bracts (or bracteole) surrounding the base. The calyx can measure to the length of about 3.2
5.7 cm, and they surround the fruit. The velvet-textured fruit pod of 1.25
2 cm length is green when immature, with
three to four seeds contained in each of the representing five valves. When fruits are mature and dry, the pods turn brown in color and rupture. Seeds appear kidney-shaped, light-brown in color, 3
5 mm long, and surrounded with miniature, stout, and stellate hairs (Mahadevan & Kamboj, 2009). All these characteristics are adaptive qualities that also guide the selection of the planting materials for crop production. Various components (calyces, leaves, and seeds) of the Roselle can be used in fresh or dry form or even can be processed into different valuable products. Generally, there are genotypic differences in these various plant parts (calyces, leaves, and stem). There is evidence that parameters such as seed weight (kg/ha), plant population (1000/ha), number of branches per plant, number of capsules per plant, hay weight (kg/ha), and plant height can contribute to the determination of the calyx yield (kg/ha). In other words, careful selection of quality seeds based on the earlier-mentioned parameters can improve plant stand, plant architecture, and eventually increase calyx yield (Atta et al., 2011). So far, the best criteria for improving Roselle plant is by selection; study outcomes prove a positive relationship between calyx yield and important parameters (including characters of seed weight, number of branches per plant, number of capsules per plant, hay weight, and plant height) (Sabiel, Ismail, Osman, & Sun, 2014).

1.3 Safe production of Roselle in climate change In most rural areas, women folks are in charge of the cultivation of minor vegetables, including Roselle. Roselle is planted mostly at the peripheries for farm demarcations or intercropped with staple crops. Harvested Roselle plants are processed into various products; the women enhance the market value to support the household income (Van Walsum, 2009). Although the Roselle crop typically grows as a perennial plant, it is mainly cultivated as an annual erect shrub and can be grown on the field for 5 months between planting and harvesting (Bailey & Bailey, 1976; Mohamed et al., 2012). The plant is susceptible to changes in day length; for that reason, it is recommended that the planting time is aligned more toward the

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proportion of the day than rainfall requirements (Mohamed et al., 2012). With the shorter days and decreased light intensity, flowering is induced, starting in September or later, depending on the growing area (Mohamed et al., 2012). Occasionally, growers in Sudan will allow seeds to ripe fully, and abscission of leaves will occur before harvesting the pods (Plotto, Mazaud, Ro¨ttger, & Steffel, 2004). Even though Roselle does very well in soils with suitable organic materials and essential nutrients and performs adequately on moderately infertile soils, it can survive in moderately high temperature during the vegetative and fruit production phases. Nevertheless, the ideal rainfall of about 45
50 cm well distributed across the 90
120 days growth period is most suitable (Adanlawo & Ajibade, 2006). It takes between 3 and 4 months for the Roselle plants to reach commercial value at maturity before harvesting the flowers. The plant is well situated in the tropics with a fairly rainfall distribution between 1500 and 2000 mm annually from sea level to almost 600 m in altitude. The plant tolerates warmer and more humid climate with nighttime temperature of 21 C and more but cannot withstand frost and fog injury. Therefore premature flowering is possible when the plant is exposed to 13 hours of sunlight in the first months of growth (Ismail, Ikram, & Nazri, 2008). The plant has a deep rooting system and requires in-depth preparation of seedbed. The seeding rate is between 6 and 8 kg/ha with about 2.5 cm planting depth. It is recommended that the seeds are sown at the start of the wet season at distances of 60
100 cm 3 45
60 cm. Larger calyces are produced when plant density is less. Sowing of seeds is by hand or use of modern grain drills. Seedling is then thinned by hand to a single plant stands to ensure appropriate plant density. There are more than 100 Roselle cultivars or varieties and the main marketable varieties are found in China, Thailand, Mexico, and Africa. In Africa, the major producing countries are Sudan, Senegal, and Mali (Ahmed, 1980; Plotto et al., 2004). In SSA between 1961 and 2016, bast fiber crop acreage moved from 15,000 to 25,000 ha, which was approximately 67% growth. Subsequently, it declined from 1.15 to 0.67 t/ha, representing 42% decrease (Tetteh et al., 2019). Sudan is currently the leading producer of Roselle in the SSA, although producers still perceive it as a hunger crop. In addition, when farmers foresee drought, they will choose to cultivate Roselle instead of cereals; the crop is hardier in extreme weather conditions (Mohamad, Nazir, Rahman, & Herman, 2002). Available data showed that in the 2000/2001 season, total land area under Roselle cultivation was approximately 121,800 ha, compared with approximately 9370
32,950 ha in the earlier years (1970s) and about 20,160
25,160 ha in the 1980s. The increase in the cultivated land area produced extra yield; thus 454 tons in the 1960s to 26,000 tons in the 1999/2000 seasons (El-Awad, 2001). In western Sudan, Roselle serves as a vital cash crop providing substantial income for the smallscale farmers. It is grown mostly under traditional farming systems with low farm inputs and exclusively handled as a rainfed crop (El Naim & Ahmed, 2010).

1.3 Safe production of Roselle in climate change

China and Thailand are involved in substantial Roselle production, making them major suppliers of the world. Higher-quality Roselle products are from Thailand; so, the country has made heavy investment in its production. Recently, Tetteh et al. (2019) also reported that India is the primary grower of Roselle in the world. Countries such as Mexico, Egypt, Senegal, Tanzania, Mali, and Jamaica are also important suppliers of Roselle products, but their production quantities can only meet the domestic market demands (Mohamad et al., 2002). Though Sudan produces the best Roselle in the world, the quality and quantities produced are limited by insufficient and poor handling and processing practices. The five major Roselle growing states in India (Andhra Pradesh, Bihar, Orissa, West Bengal, and Maharashtra) cultivated a total area of 84,000 ha between 2012 and 2013 (Sen & Karmakar, 2014). Satya, Karan, Kar, Mahapatra, and Mahapatra (2013) reported that Roselle jute production is only 0.55% of the cropped area in India but supports as much as 4 million farm folks, 0.25 million workers in the manufacturing industries, and 0.50 million direct traders. Roselle in Senegal is locally processed into drinks, and the local industries use a small percentage of calyces. Here, the red H. sabdariffa in Senegal in 2012 was 2885 tons and only 200 tons of was used by the local agroprocessors (Cisse et al., 2009). Generally, there have been unlimited discussions on the safe production of vegetable crops regarding water management, synthetic pesticides use, and fertilization practices. Characteristically of most vegetable crops, Roselle is relatively responsive to poor production practices. In cases where the plant is grown in different agroclimatic places than the regions of natural origin or most conducive environment, it becomes more vulnerable to adverse soil conditions and climatic factors with significant potential yield losses. For that matter, the plants will need special care and resources to make up for the deficits. Climate change factors, including fluctuating in temperatures, shortage of available water for irrigation or drought situation, the rate of regular to prolonged flooding, changes to extreme pH levels, and wind velocity increases, culminate in creating unsustainable vegetable farming conditions (Singh & Bainsla, 2014). Roselle plant will respond to severe environmental stress diversely according to its genotype and other crop factors (Singh et al., 2013). There is evidence that vegetable yields in tropical areas can persistently be low due to genotypic or environmental effects or their interactions (Singh et al., 2013). An estimate of 50% vegetable crop yield loss was found to be mostly due to environmental stresses (Bray, 2002), and certainly revenue from major farming choices will also be affected (Singh & Bainsla, 2014). Over time, more and more strategies are being pursued to adapt to the potential impact of climate change (Phophi & Mafongoya, 2017; Srang-iam, 2011; Sto¨ber et al., 2017). It requires multifaceted techniques to attain a sustainable outcome of crop productivity. For instance, diversity in parent materials has been harnessed for crop improvement. The diversity in parent materials will each time in a hybridization program produce positive effects; this means that identification of qualities that determine the total diversity of genotypes among the populations

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must be done well (Mehetre, Mahajan, Patil, & Hajare, 1994). The success of improving any crop depends on the nature and scope of genetic variability available in the plant (Satyanarayana et al., 2015).

1.4 Potential sources of heavy metals and microbial contamination of Roselle Over the years, pesticides have been used to improve crop yields, and the annual demand for chemical products in agriculture has been increasing significantly. Pesticides use has diverse detrimental effects on flora, fauna, as well as the environment. More so, consumers are exposed to high chemical risk in food. There are various ways that heavy metals from pesticides and microorganisms can contaminate Roselle (Fig. 1.1). Soil amendments with organic and inorganic fertilizers have also proved to be potential sources of heavy metals, making them bioavailable for uptake into edible parts of the plant (Chaney, 2012). Continuous crop cultivation, especially in the tropics, causes soil depletion of productive capacity, hence the need for constant replenishment with fertilizers. The increased application of organic fertilizers such as poultry manure (Delgado, Mrialles, Peralla, & Almestre, 2014) and municipal solid waste compost (Ghaly & Alkoaik, 2010) also predispose treated plants to heavy metal contamination. Polluting the air with lead (Pb) and cadmium (Cd) by vehicles will eventually find their way into the soil through precipitation and affect plant life (Popescu, 2011). In similar studies, Abubakari, Moomin, Nyarko, and Dawuda (2017) found Pb (0.8 mg/kg) and Cd (5.0 mg/kg) concentrations in Roselle leaves treated with composts to be above the maximum residue levels (MRLs) of Cd (0.2 mg/kg) and Pb (0.3 mg/kg) established by the European Commission and Codex Alimentarius Commission. It was also reported that some leafy vegetables cultivated in urban environments were contaminated with Pb levels above the MRLs (Wamalwa et al., 2015). Other sources of Roselle contamination include the use of synthetic pesticides to control plant pests. Pimentel and Levitan (1986) indicated that less than 0.1%

FIGURE 1.1 Flow chart of potential sources of Roselle heavy metal and microbial contamination on human and animal health.

1.5 Conclusion

of these pesticides are applied to reach the target pests, which means the leftover pesticides find their way into the environment to contaminate soil, water, and plants. An estimated 200,000 acute poisoning deaths occur annually due to pesticide abuse (Svensson et al., 2013), and 99% of these occurrences have been noticed in developing countries (Goldman, 2004). In the past few decades, pesticide use has increased astronomically, with most of the weaker and less firm pragmatic environmental regulations being implemented in developing countries. The availability of water determines the plant survivability and the existence of beneficial microorganisms. In recent years, the increasing freshwater shortage is a significant challenge in crop production, especially in urban areas but more critical in the semiarid and arid regions. Consequently, farmers resort to the use of untreated wastewater as a common practice to irrigate vegetables. However, the consumption of raw leafy vegetables without appropriate decontamination of microbial load (Total coliforms, Escherichia coli) may induce adverse human health risk (Hussain & Qureshi, 2020). A study conducted by Ataogye (2012) in the Upper East Region of Ghana found that the microbial load on Roselle leaves (both dry and fresh) were higher than that of World Health Organization and the International Commission on Microbiological Specifications for Foods standards. Here, levels of total and fecal coliforms, Enterococci and E. coli were measured on both fresh and dry leaves. The study outcome indicated that decontaminated water for irrigation would reduce microbial load and lessens human health risk. Generally, vegetables are very vulnerable to chemical contamination, and they are known to be high accumulators of heavy metals, representing a high risk to consumers who frequently depend on these foods. The risk of consuming heavy metal-contaminated vegetables includes chronic health complications such as damage to the liver and kidneys (Mahmood & Malik, 2014). Finally, pragmatic actions to build composed and practical agroecosystems guarantee persistent increasing yields of Roselle; thus it also offers greater capacity to endure changes in climate and survive the possible occurrence of pests and diseases resistance. For instance, natural plant extracts as biocontrol methods against plant enemies, the use of varieties that are tolerant to drought, sustainable water and soil conservation methods (mulching and crop rotation), reduction of emissions of greenhouse gasses, and provision of carbon sinks have been recommended. These methods have been stated as the most effective technologies to mitigate the negative impact of climate change, misuse of synthetic pesticides, and fight food insecurity.

1.5 Conclusion In summary, Roselle plants are very suitable for tropical areas, thus adapted to hot climates. For many decades it has been used as herbal medicine in phytotherapy and nutritious vegetable. The Roselle crop is appreciated for its multipurpose

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bast fiber and considered an important commodity marketed worldwide. Large genetic variability in Roselle characterized by vast differences in morphotypes provides information for the development of improved cultivars (such as fiber yield and quality) and conservation of desired traits. Various methods to manipulate the Roselle plant or its environment to increase yield quality have been investigated. The selection of Roselle according to phenotypic qualities can increase calyx yield, but the use of molecular markers helps to conduct accurate characterization of the plant species. Finally, sources of product contamination have received massive consideration first to ensure the safety of Roselle products for consumption.

Acknowledgment The author is thankful to the University Development Studies, Ghana, for providing facilities to undertake this chapter review work.

Conflict of interest The author has declared that no conflict of interest exists.

References Abubakari, M., Moomin, A., Nyarko, G., & Dawuda, M. (2017). Heavy metals concentrations and risk assessment of Roselle and jute mallow cultivated with three compost types. Annals of Agricultural Sciences, 62(2), 145
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557. Ahmed, A. (1980). Karkade (Hibiscus sabdariffa L.) seed as new oilseed and a source of edible oil. University of Reading. Alves, C., Silva, A., Reis, L., Freitas, M., Rodrigues, L., & Alves, D. (2010). Ecodesign of automotive components making use of natural jute fiber composites. Journal of Cleaner Production, 18(4), 313
327. Ankrah, N. A., Tetteh, A. Y., Coffie, N., & Niagiah, A. (2018). Characterization of Roselle (Hibiscus sabdariffa L. var. altissima Wester) accessions in Northern Ghana by agromorphological traits. Journal of Agricultural Science, 10(9). Ataogye, G. (2012). Microbial contamination of an indigenous leafy vegetable, Roselle (Hibiscus sabdariffa L.) and associated risk factors on farm and market samples in the Kasena-Nankana East Municipality of the upper east region. Atta, S., Seyni, H. H., Bakasso, Y., Sarr, B., Lona, I., & Saadou, M. (2011). Yield character variability in Roselle (Hibiscus sabdariffa L.). African Journal of Agricultural Research, 6(6), 1371
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Osman, M., Golam, F., Saberi, S., Abdul Majid, N., Nagoor, N. H., & Zulqarnain, M. (2011). Morpho-agronomic analysis of three Roselle (’Hibiscus sabdariffa’L.) mutants in tropical Malaysia. Australian Journal of Crop Science, 5(10), 1150. Phophi, M. M., & Mafongoya, P. L. (2017). Constraints to vegetable production resulting from pest and diseases induced by climate change and globalization: A review. Journal of Agricultural Science, 9(10), 11. Pimentel, D., & Levitan, L. (1986). Pesticides: Amounts applied and amounts reaching pests. Bioscience, 36(2), 86
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Srang-iam, W. (Ed.). (2011). Fighting global climate change, securing local livelihood: The paradox of carbon reduction and agricultural vulnerability in Thailand. Colorado Conference on Earth System Governance: Crossing Boundaries and Building Bridges. Sto¨ber, S., Chepkoech, W., Neubert, S., Kurgat, B., Bett, H., Lotze-Campen, H. (2017). Adaptation pathways for African indigenous vegetables’ value chains. In Climate Change Adaptation in Africa: Springer, 413-433. Svensson, M., Urinboyev, R., Wigerfelt Svensson, A., Lundqvist, P., Littorin, M., & Albin, M. (2013). Migrant agricultural workers and their socio-economic, occupational and health conditions
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167). Bourton on Dunsmore, UK: Practical Action Publishing. Wamalwa, K., Lusweti, J., Lutta, S., Anditi, B., Imbila, O., & Muthoka, T. M. (2015). Variation of pollutant levels in vegetables: A case study of Kitale municipality, TransNzoia County, Kenya. African Journal of Education, Science and Technology, 2(2), 146
160. Wilson, F., & Menzel, M. Y. (1964). Kenaf (Hibiscus cannabinus), Roselle (Hibiscus sabdariffa). Economic Botany, 18(1), 80
91.

CHAPTER

Harvesting, storage, postharvest management, and marketing of Hibiscus sabdariffa

2

Mildred Osei-Kwarteng1, Joseph Patrick Gweyi-Onyango2 and Gustav Komla Mahunu3 1

Department of Horticulture, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana 2 Department of Agricultural Science and Technology, Kenyatta University, Nairobi, Kenya 3 Department of Food Science and Technology, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana

Chapter Outline 2.1 Introduction ................................................................................................... 16 2.2 Harvesting of Roselle ...................................................................................... 17 2.2.1 Determinants of harvest maturity of Roselle .....................................17 2.2.2 Harvesting of leaves and tender shoots ............................................18 2.2.3 Harvesting of calyces .....................................................................19 2.2.4 Harvesting of seeds .......................................................................22 2.2.5 Harvesting stems for fiber ..............................................................22 2.3 Postharvest management of harvested Roselle ................................................. 23 2.3.1 Postharvest management of leaves and tender shoots .......................23 2.3.2 Postharvest management of calyces ................................................23 2.3.3 Postharvest management of seeds ..................................................25 2.3.4 Postharvest management of stem (fiber) ..........................................26 2.4 Storage of useful parts of Roselle .................................................................... 27 2.4.1 Storage of leaves and tender shoots after harvest .............................27 2.4.2 Storage of harvested calyces ..........................................................27 2.4.3 Storage of harvested seeds .............................................................27 2.5 Marketing of Roselle produce and products ..................................................... 28 2.5.1 Marketing of leaves and tender shoots, calyces, seeds, and fiber .......28 2.6 Conclusion ..................................................................................................... 30 References ............................................................................................................ 30

Roselle (Hibiscus sabdariffa). DOI: https://doi.org/10.1016/B978-0-12-822100-6.00010-0 © 2021 Elsevier Inc. All rights reserved.

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CHAPTER 2 Harvesting, storage, and management of Roselle

2.1 Introduction Hibiscus sabdiriffa var. sabdariffa, commonly known as Roselle, is an annual herbaceous plant that grows in tropical and subtropical countries (Plotto, Mazaud, Ro¨ttger, & Steffel, 2004). It is among the plants with several useful products depending on plants’ parts in question and processing approaches. Common products derived from this plant comprise herbal medicine, food (sauce and vegetable salads from leaves, fruit salads from calyx), soft drinks (juices), syrup, jelly, jam, butter, tea, wine, cake, ice-creams, perfumes, marmalade, seasoning products, fibers, spices, sauces, and desserts (Duke, 1983; Mohamed, Sulaiman, & Dahab, 2012; Plotto, Mazaud, Ro¨ttger, & Steffel, 2004). According to FAO (2004), it is one of the plants with the highest volume of specialty botanical products. The useful parts include the leaves, calyces, stems, and seeds. However, the calyx is the Roselle plant part that has high economic value (Gomaa & Rashed, 2016). Roselle plant requires about 4
6 months from sowing to harvesting. However, the growth and development of this plant are strongly influenced by environmental conditions and genotype (Haryati, Nisa, Julianti, & Blunden, 2018). Hence, harvesting of the whole plant or any useful part of the plant is dictated by the growth environment among other factors. The first harvest is usually at thinning where young seedlings are collected. Later, when the plant is 6
8 weeks old, branches of about 50 cm long can be picked thrice during the vegetative growth (Mcclintock & Tahir, 2004). On the other hand, the other useful parts such as the calyces, flowers, seeds, and stems are harvested at later stages of the plant development and quality characteristics for these parts determine their harvest maturity. There are several methods used in storing harvested Roselle produce, some vary from one country to the other where Roselle production and marketing are common. For instance, in Niger, harvested seeds/grains can be stored in nonhermetic bags/containers such as woven bags, plastic jugs, plastic bags, and granaries. There are some preservatives added to Roselle produce and comprise wood ash, neem compounds, sand, and insecticides to prevent pathogen attack (Boureima, Moussa, & Lowenberg-DeBoer, 2015). Postharvest management involves the handling, storage, and transport of harvested Roselle produce and products. These processes are key to improve the quality, durability, safety, and marketability of Roselle produce and products. For example, common postharvest activities for Roselle calyces may include shelling, drying, packaging, and storage (Gomaa & Rashed, 2016). However, some of the useful parts of Roselle are marketed fresh in bunches (tender shoots and leaves), while others undergo simple processing procedures such as drying, especially the flowers, calyx, and seeds sold in local markets. In such cases, postharvest procedures are simple. In regional and international markets, Roselle products may be sold as processed creams, juices, cycles dried, and packaged in bales and in several other processed forms. Marketing forms of Roselle products vary depending on the producing countries as well as by the specifications from the importing countries.

2.2 Harvesting of Roselle

Common producers supplying large international markets comprise Malaysia, Sudan, Egypt, Thailand, China, and Mexico, while the major importers comprise the United States and Germany (Plotto, Mazaud, Ro¨ttger, & Steffel, 2004). This chapter provides an elaborate discussion on the harvesting, storage, postharvest management, and marketing of some useful products of Roselle. The main useful parts to discuss are the leaves or tender shoots, calyces, flowers, seeds, and stems.

2.2 Harvesting of Roselle Harvest maturity is categorized as either physiological or horticultural. Physiological maturity is the stage at which the useful organ/plant part (e.g., fruits) completes its development. Physiological maturity is normally used for fruits, while horticultural maturity is more of vegetables (mostly leafy vegetables). Horticultural maturity is the stage of development when a plant or a plant part possesses the required attributes (peak acceptable quality) for utilization by the consumer for a particular purpose. The timing of harvesting for a useful part of Roselle is dictated by the determinants of harvest maturity of the useful part. Also, the purpose of production determines which useful part and when this part would be harvested during the production season. For example, harvesting of stems for fiber requires a long period of development, this was observed to be about 6 months in some parts of India (Agarwal & Dedhia, 2016). Different parts of Roselle are harvested at different stages of development. Roselle grown for calyces and seeds is harvested after completion of the flowering stage of development. Flowering of Roselle is induced during short-day periods when the light intensity reduces, this in most countries coincides with September or later (Plotto, Mazaud, Ro¨ttger, & Steffel, 2004). In tropical parts of East and Western Africa, harvesting of Roselle plant for calyces starts in November. It is necessary to harvest Roselle plant parts at the appropriate maturity stage for highquality produce (sensory, nutritional, and other consumer-preferred qualities) as well as to reduce postharvest losses.

2.2.1 Determinants of harvest maturity of Roselle Determinants of harvest maturity of the useful parts of Roselle are necessary to ensure that these parts (1) can be harvested at the best/maximum quality stage (appearance, texture, flavor, and nutritive value); (2) are harvested at the optimum stage at which senescence can be delayed and the shelf life prolonged; (3) are harvested at a stage where returns can be maximized; and (4) can be transported for long-distant marketing. Different Roselle varieties respond differently to growth conditions and would therefore reach peak maturity period for different harvestable parts at different

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CHAPTER 2 Harvesting, storage, and management of Roselle

Table 2.1 Time to maturity of Roselle parts observed by different researchers. Country

Roselle part

Harvesting time after planting

Reference

India

Stems

180 months

Agarwal and Dedhia (2016)

Egypt

Calyces

180
183 days

Gomaa and Rashed (2016)

Nigeria

Calyces

100
160 days

Nnebue et al. (2014)

Tanzania

Calyces

4
5 months

ITM (2008)

Ghana

Calyces

About 3 months

Amoasah, Appiah, and Kumah (2018)

Sudan

Calyces

About 181 days

Mohamed et al. (2012)

periods of development. Roselle plants are highly affected by environmental conditions such as humidity, soil fertility, diseases, and photoperiod; these conditions would significantly influence growth, maturity, and yields. For example, for cases of high humidity at flowering, early harvesting of cycles may be profitable since it would avoid serious pest attacks. Planting density also has an influence on the development of Roselle plants. Due to high dependence on photoperiod, planting dates have also a considerable influence on the period taken by Roselle to mature, for instance, in parts of Nigeria, Roselle plants grown in June and July reach flowering time faster than those planted in May (Castro et al., 2004; Nnebue et al., 2014). On the other hand, planting in May exposes Roselle plants to extended periods of long day; this would be suitable for leaves and stem production (Nnebue et al., 2014). The method of planting and field conditions may also have an influence on the period taken to maturity, since plants that are directly sowed on the field may develop differently from those grown in nurseries and later transplanted. But Roselle cultivation is mostly propagated by direct field sowing. Direct field establishment may lack uniformity; therefore such plants require thinning during seedling development, this will encourage removal of poorly developing seedlings. Roselle plants grown for different purposes in diverse countries have been observed to take about 4
6 months to mature (Table 2.1) (Agarwal & Dedhia, 2016; Gomaa & Rashed, 2016; ITM, 2008; Nnebue et al., 2014).

2.2.2 Harvesting of leaves and tender shoots Leaves and shoots are harvested for use as salads and to be prepared as vegetables. They are harvested when young and tender by hand (Fig. 2.1). These young and tender leaves and shoots can be prepared alone or in a combination of other vegetables. There are cultivars of Roselle that produce large quantities of leaves compared to calyces, especially the ones propagated vegetatively. The yield of

2.2 Harvesting of Roselle

FIGURE 2.1 Tender leaves and shoots of Hibiscus sabdariffa.

leaves may be up to 10 t/ha (Tindal, 1983). Leaves may be harvested and used as fodder while still fresh as indicated by Plotto, Mazaud, Ro¨ttger, & Steffel, 2004. Fresh leaves yield can be about 10 t/ha; however, there are greater variabilities in cultivars as well as the environmental conditions of the site of cultivation.

2.2.3 Harvesting of calyces The calyces ripen about 3 weeks after the start of flowering, about 100
160 days after transplanting (Tindal, 1983). At this time, the calyces are fleshy, flowers have dropped, and the seeds are ripe but not yet exposed since the pods are still closed (Fig. 2.2). Calyces mature at about 20 days following flowering, this is normally about 2
3 weeks after flowering when the flowers have dried up and fallen off. However, harvesting of calyces is done when they are still fresh before they dry up. At this time of harvesting, calyces would still have high moisture content (85%) (Mcclintock & Tahir, 2004; Shruthi et al., 2016). Optimum period of harvesting calyces varies depending on the climate. For instance, in Bangladesh, the optimum

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CHAPTER 2 Harvesting, storage, and management of Roselle

FIGURE 2.2 Mature Roselle fruit: (A) the fleshy calyces; (B) green seed capsule.

FIGURE 2.3 Tools used for harvesting Roselle: (A) hand clippers and (B) knife.

time for harvesting fresh calyces was observed to be about 45 days after anthesis, while in Indonesia, this was observed to be about 33 days after anthesis (Haryati et al., 2018). Harvesting of calyces is done by picking or plucking off mature Roselle fruits from the plants. This can be done manually by hand or by use of clippers, sometimes knives can be used in harvesting and shelling (Fig. 2.3). Late harvesting of calyx is entirely by use of clippers because at such time they are a little dry and difficult to handpick. Furthermore, late harvesting of calyx makes them susceptible to disease attacks and deterioration by sun cracking (Morton, 1987; Plotto, Mazaud, Ro¨ttger, & Steffel, 2004). Regular picking of the fruits encourages prolonged flowering, this increases the productivity of Roselle plants. Roselle fruits are temporally carried in buckets and sacks such as gunny sacks before being taken to the shelling or deseeding centers. Transfer to the deseeding centers can be done using refrigerated trucks especially when it is located far away from the farm, but for those in proximity to the farms the fruit can be carried in buckets and sacks. In large plantations, such as those in Malaysia, the fruits are collected in buckets and sacks then transferred to larger baskets or crates before ferrying to the deseeding centers using large refrigerated lorries (Ali, Kamarudin, Me, Sulaiman, & Alli, 2019). At these centers, the first activity is to remove the seed capsules before further processing (Fig. 2.4). Each fruit yields about 7
10 g of calyx, and at maturity the average weight of calyces per fruit is about 11
12 g (Haryati et al., 2018). A yield of 1.5 kg per plant

2.2 Harvesting of Roselle

FIGURE 2.4 Ripe Roselle calyces after removal of seed capsules: (A) green Roselle calyces, (B) light red Roselle calyces, and (C) dark red Roselle calyces (Hussein, Shahein, El Hakim, & Awad, 2010). From Hussein R. M., Shahein Y., El-Hakim A. E., & Awad H. M. (2010). Biochemical and molecular characterisation of three coloured types of Roselle (Hibiscus sabdariffa). Journal of American Science, 6(11), 726
733. https://doi.org/10.7537/marsjas061110.105.

FIGURE 2.5 Deseeding tools for Roselle fruits: (A) traditional deseeding tool and (B) SORREL deseeding tool. From Ali, E. A., Kamarudin, K. M., Me, R. C., Sulaiman, R., & Alli, H. (2019). The development of SORREL systematic Roselle harvesting system for efficient transfer process. IOP Conference Series: Materials Science and Engineering, 697. https://doi.org/10.1088/1757-899X/697/1/012029.

can be obtained for the calyces, which is approximately 8 t/ha (Tindal, 1983). Fresh calyx yields can range from 4 to 6.5 t/ha at about 12% moisture content. Several methods are used in deseeding; this can be done by use of knives or simple hand tools. There are efforts to develop new systems and technologies to enhance efficiency in harvesting of Roselle calyces. In Malaysia plantations, to improve the harvesting and handling calyces, there has been development of the SORREL harvesting system that comes with new harvesting bags (SORREL harvesting bags), SORREL harvesting, and deseeding tools; these items are more efficient than the traditional harvesting using buckets and simple tools (Fig. 2.5). These new developments also enhance maintenance of high quality for the harvested calyces (Ali et al., 2019).

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CHAPTER 2 Harvesting, storage, and management of Roselle

2.2.4 Harvesting of seeds Roselle seeds are obtained from harvested mature Roselle fruits. During harvesting of the mature fruits, the inner seedpod is normally green and closed. This is always the case for Roselle grown for fleshy calyx. However, in other cases the fruit can remain on the plant to dry up. This would allow the seeds within the seedpod to reach maximum maturity. After harvesting of the fleshy Roselle fruits, the processes of obtaining the seeds involve removal of calyces that separates the calyx from the seedpod. Simple hand tools are used for this procedure. The obtained ripe seeds can be further dried and stored. Roselle plants grown for seeds may be managed differently by allowing the fruits to partially dry on plants before picking the Roselle fruits. The dry calyx easily cracks by exposure to sunlight (Fig. 2.6). Some of the Roselle cultivars open when mature, this can enhance removal of the ripe seeds. Delayed harvesting of fruits may also enhance improved quality of the harvested seeds. The weight of seeds per single Roselle fruit is about and 2
3 g (Haryati et al., 2018) (Fig. 2.6).

2.2.5 Harvesting stems for fiber Cultivation of Roselle for fibers is common in India. Harvesting of stems for fiber is done by cutting the stems at about 4
5 months before flowering to prevent the decline of fiber quality after flowering (Mcclintock & Tahir, 2004). In some other regions due to variable growth conditions, harvesting of stems for fibers may be carried out after a longer period, for example, harvesting of stems for fibers in Maharashtra, India, is done after 6 months (Agarwal & Dedhia, 2016).

FIGURE 2.6 Hibiscus sabdariffa seedpod containing mature seeds.

2.3 Postharvest management of harvested Roselle

2.3 Postharvest management of harvested Roselle There has been less research in development of systems and tools to support postharvest handling of Roselle produce. For example, harvesting of the Roselle parts is manual, where simple hand tools are used, such as the hand clippers and shelling tools. Development of harvesters may be easier for stems compared to fruits that display an indeterminate pattern of development and maturity, spanning an extended period on a single plant.

2.3.1 Postharvest management of leaves and tender shoots Harvested young leaves and tender shoots are used in salads or cooked as greens together with other vegetables or with meat and fish. Juices can also be made from strained boiled young leaves and tender stems (Plotto, Mazaud, Ro¨ttger, & Steffel, 2004).

2.3.2 Postharvest management of calyces The initial procedure for postharvest processing of Roselle calyces is shelling. Shelling is a process of removing seedpods from the Roselle calyces. This is done by hands or by simple hand tools (Gomaa & Rashed, 2016). Although using hand tools in shelling may be faster and less labor intensive, it may result in lower fresh yield of calyces. After separation of calyx from the seedpods, the fresh Roselle calyx is further processed using different methods, which can comprise cooking and fermentation to produced juice, jelly, and Roselle syrups or sun drying of the calyx. For juices and jelly the process may also involve heating to varied temperatures and addition of sugars as well as other compounds such as preservatives. Sometimes, the calyces can be left to dry on the Roselle plant, this reduces the need for drying after harvesting. However, for the fleshly harvested mature fruits, the calyces after separation of pods through shelling can be air dried, this should be undertaken under the shed and in a well-ventilated environment. This allows for the removal of excess moisture from the calyces. When sun dried, the quality of calyces would deteriorate and will lower their market price. Furthermore, during drying, there is a need for caution to avoid contamination. In this light, to avoid contamination with the soil during drying, plastic sheets are placed on the ground. Also, the plastic sheets help to keep a good value product. Calyces can be dried by direct sunlight, using solar panels, solar driers, ovens, or by use of dehumidifiers (Amoasah et al., 2018; Gomaa & Rashed, 2016; Saeed, 2010). Therefore the process can use solar energy and other heat energy sources, but the heat should be relatively low, since higher temperatures with low regulation of relative humidity may result in degradation of phytochemicals. In advanced systems such as ovens, higher temperatures are used, for example, temperature of

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CHAPTER 2 Harvesting, storage, and management of Roselle

about 55 C has been used in oven drying for 36 hours to a moisture level of about 11% without affecting the quality of calyces (Gomaa & Rashed, 2016). The common methods of drying calyces include forced air oven drying (60 C), constant temperature and humidity chamber (i.e., drying at 35 C, 45 C, 55 C, and 55 C at a relative humidity of 30%, 35%, 40%, 45%, and 50%), tunnel drying (drying calyces at 70 C for 4.5 hours), thin layer drying (tray drying; drying at temperature 40 C, 50 C, and 60 C and air velocity of 1.5
1.6 m/s), sun and oven drying (sun drying for 3 days and oven drying for 24 hours at 50 C), hot air drying (60 C), and oven drying (oven drying at 105 C temperature for 45
60 minutes). In Sub-Saharan Africa, drying by artificial heating is rare and capital-intensive (Mcclintock & Tahir, 2004). For example, in Egypt, calyces are traditionally sun dried. In this traditional drying, calyces are spread on mats and plastic sheets laid on ground surface; this is done for about 6
10 days and results in about 14%
16% moisture content, suitable for storage. However, sun-drying method results in heterogenous quality of the calyces products; this may have an effect on the marketability (Gomaa & Rashed, 2016). The above processing procedures, involving drying, heating, and fermentation, enhance the quality of Roselle produce and improve the shelf life of produced products. For example, some of the processed Roselle such as juice may be used for an extended period. Dried calyces have increased shelf life because they are less susceptible to deterioration (Shruthi et al., 2016). Dried calyces are packed in woven polyethene bags in bales of about 50 kg. Extracts from calyces are obtained at varied temperatures, this has been reported to range from 25 C to 100 C (Bhat, Sridhar, & Yokotani, 2007). Dried calyces may be used later to produce hibiscus tea, a sweet herbal tea. Prior to processing, the dry calyces are package in gunny bags and transported to processing industries (Fig. 2.7). Processed tea is packaged in small packets. In terms of quality, different drying methods may have an effect on the quality of calyces; oven

FIGURE 2.7 Drying methods for Roselle calyces: (A) solar drier and (B) oven. From Amoasah, B., Appiah, F., & Kumah, P. (2018). Effects of different drying methods on proximate composition of three accessions of Roselle (Hibiscus sabdariffa) calyces. International Journal of Plant & Soil Science, 21(1), 1
8. https://doi.org/10.9734/ijpss/2018/38550.

2.3 Postharvest management of harvested Roselle

FIGURE 2.8 A flowchart of postharvest processing of Roselle calyces.

drying was observed to yield high quality of calyces in terms of the quantities of total soluble solids (Gomaa & Rashed, 2016). It has been observed that the air temperatures are very important during drying of Roselle calyces; during drying of calyces in a dehumidifier drying system, an increase in drying air temperatures from 35 C to 65 C reduces the drying time, while at low temperatures of about 35 C, increase in the drying air velocity could still reduce the drying time (Saeed, 2010). Different methods of drying have an influence on the quality of calyces (Fig. 2.8). High fat, ash, and carbohydrates were obtained in oven-dried calyces compared to both direct sun and solar-dried calyces, while solar-dried calyces were observed to have a high content of proteins (Amoasah et al., 2018).

2.3.3 Postharvest management of seeds Seeds are obtained from the seedpods or capsules by opening the capsules, the seedpod/capsule can be crushed when dry to remove the seeds (see summary in Fig. 2.8). This is followed by drying of the seeds to a moisture content of about 11%, under a shade. To obtain clean seeds, winnowing is done to remove seedpod materials (Nyarko, Bayor, Craigon, & Suleimana, 2006). Seeds to be used as planting materials are left to mature and dry within the seedpods inside the calyx. Those obtained from harvested fruits may be air dried before further storage (Fig. 2.9). Some of the harvested seeds after drying may be further processed as aphrodisiac coffee substitutes, this involves roasting of such seeds (Mohamed et al., 2012). Roselle seeds are sometimes powdered for use in food (Plotto, Mazaud, Ro¨ttger, & Steffel, 2004). Since they contain large quantities of oils (about 17%), they may

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CHAPTER 2 Harvesting, storage, and management of Roselle

FIGURE 2.9 A flowchart of postharvest processing of Roselle seeds.

be processed to produce vegetable oils (Mohamed, Fernadez, Pineda, & Aguilar, 2007). Just before oil extraction, cleaning of the seeds is necessary to remove foreign materials and other contaminants that can be achieved through washing with clean water. Afterward, the seeds should be dried before further processing. Oil extraction can be undertaken through traditional oil extraction methods from seeds or modern industrial procedures. It has been observed that after oil extraction, the oils from Roselle seeds should be stored in clean containers. It has been observed that storage of Roselle oils in dark airtight conditions preserves their quality without addition of preservatives for about 16 weeks with high maintenance of their quality (Nkafamiya, Atiku, Akinterinwa, & Fari, 2017). These oils can be used in soap and other cosmetic industries or in production of paints.

2.3.4 Postharvest management of stem (fiber) Harvested stems are subjected to retting to obtain the fibers, this process can be done through simple traditional procedures such as water retting, microbial retting, or through advanced industrial processes (Agarwal & Dedhia, 2016). In water retting, harvested stems for fiber are soaked in water for about 2 weeks and the bark is removed after the 2 weeks. To separate the fiber, stems are then beaten, washed, dried, and sorted based on length, color, stiffness, and purity (Mcclintock & Tahir, 2004). The fibers from Roselle stems can be used to make bags, clothes, linen, ropes, fishing nets, and other items.

2.4 Storage of useful parts of Roselle

2.4 Storage of useful parts of Roselle Majority of Roselle farmers undertake small production, they have low investment in value addition and processing procedures; therefore they face challenges in marketing their produce (Boureima, Moussa, & Lowenberg-DeBoer, 2015). Just like many other crops, due to seasonality of the crop, there are many instances of high market fluctuations occurring during peak harvest seasons. Most of the produce is sold shortly after harvest with less storage periods.

2.4.1 Storage of leaves and tender shoots after harvest Most leaves and tender shoots harvested are used while fresh, a substantial amount is used for local consumption by farmers’ families and their communities. Harvesting is done in small quantities and handling is done in ways similar to other vegetables. They are carried in well-ventilated containers and cooked shortly after harvesting with less storage requirements. For the leaves used in extracts production, these may be carried through cooking or other juice extraction procedures, the juice is processed with addition of preservatives and packaged in containers for extended storage.

2.4.2 Storage of harvested calyces Dried Roselle calyces can be stored over a period of 1 year without losing their quality (Gomaa & Rashed, 2016). Extracts from calyces can be packed in polyethene and glass bottles or polythene sachets. This allows storage of these extracts for extended periods of time without deterioration of their quality (Gomaa & Rashed, 2016). Cold storage of freshly harvested calyces can also prolong the shelf life with less deterioration. According to Kek et al. (2017), storage of fresh Roselle calyces at 10 C for 8 days did not result in significant weight loss, color deterioration, or softening compared to storage at room temperature (23 C). On the other hand, storing the calyces’ extracts at 4 C maintains the quality of extracts for a period of about 3 months (Moussa, Ndeye, Joseph, & Mady, 2018). Dried calyces can be powdered and combined with other compounds or encapsulated before storage. For example, powdered Roselle calyces can be encapsulated with mesquite gum before storage, this has been shown to extend the storage time to about a year with a minimum reduction in quality especially under cold storage (Ochoa-Velasco, Salazar-Gonza´lez, Cid-Ortega, & Guerrero-Beltra´n, 2017).

2.4.3 Storage of harvested seeds Roselle seeds are stored in woven polypropylene bags, plastic bags, granaries, metal drums, purdue improved crop storage (PICS) triple bags, old clothes, fabrics, bowls, and even plastic jugs. There is challenge of extended storage of seeds

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CHAPTER 2 Harvesting, storage, and management of Roselle

due to susceptibility to pest attacks so farmers use various methods to manage these pests during storage in diverse ways. In Niger, farmers mix seeds with sand, wood ash, or and neem leaves to prevent the seeds from pest attack (Boureima, Moussa, & Lowenberg-DeBoer, 2015). To avoid the rapid deterioration of stored Roselle seeds, appropriate storage containers are supposed to be used; in addition, there should be measures to prevent damage from moisture and storage pests. In Ghana, the use of polyethene bags, bottles, and plastic containers was found to be suitable for long-term storage of Roselle seeds especially alongside the use of charcoal as a desiccant (Nyarko et al., 2006). Other containers used in storage of Roselle seeds comprise earthen pots and gourds, but these traditional containers are less suitable for long-term storage due to significant reduction in quality that affects germination. The practice of using antipest compounds in seed storage, especially use of powders obtained from neem and wood ash, is common for seed storage among Roselle farmers, but Nyarko et al. (2006) observed that these powders did not have a significant effect on the germination capacity of Roselle seeds after 1 year of storage, it is possible that they have limited protection or rather there were no storage pests for Roselle seeds for this case.

2.5 Marketing of Roselle produce and products Roselle markets are local, regional, and international. Huge international markets are in the United States and Germany and they are dominated by major producers such as Malaysia, China, and Thailand. Global Roselle market is poorly regulated and hence faces dramatic price fluctuations. The supply of fresh Roselle produce (leaves and cycles) is high due to high production from various parts of the world, which results in oversupply. At times, poor prices lead to crop abandonment by some farmers. Prices and marketing quality are influenced by Roselle products’ taste, color, and cleanness among other attributes. However, in most Roselle-producing countries, there are several local markets. For example, Kenyan farmers process the herbal tea and beverages which they sell to local supermarkets, shops, and other retailers specialized in herbal products. The east Africa Roselle Company produces juice and hibiscus tea. Hibiscus beverages are commonly produced for locals in many countries including Egypt, Sudan, Nigeria, Mexico, West Indies, Senegal, Benin, Coˆte d’Ivoire, Burkina Faso, Tanzania, and Kenya. Among regional markets, Kenya imports hibiscus from Tanzania for flavoring and other value addition procedures.

2.5.1 Marketing of leaves and tender shoots, calyces, seeds, and fiber The useful products of Roselle are sold locally in countries where they are produced and exported internationally to the United States and Germany. China and

2.5 Marketing of Roselle produce and products

Thailand are the world-leading producers of Roselle with Thailand providing high-quality Roselle products in the international market. Nevertheless, the world’s best quality Roselle comes from Egypt and Sudan, although the quantity is low and poor processing hampers the quality (Plotto, Mazaud, Ro¨ttger, & Steffel, 2004). Fibers from stems are used in the production of clothes, linen, nets, ropes, and paper; however, fibers from Roselle stems face competition from other fibers in market such as those from other plants and synthetic fibers as there is a very small market share for Roselle, and this affects marketability. In addition, Roselle grown for fiber takes a long period to mature; this is another discouragement to their production for this purpose.

2.5.1.1 Marketing of leaves, tender shoots, and calyces Roselle shoots are sold locally in bunches in African markets (Mcclintock & Tahir, 2004). Leaves are commonly cooked as vegetables in Sudan and Malaysia. In West Africa, dried calyces are sold in bulk or in individual sachets (Mcclintock & Tahir, 2004). Also, leaves can be cooked in groudnut soup, commonly referred to as “Bra” or “Bito” soup in Northern Ghana. In Ghana, dried calyces are sold in the market in basins or bigger bowls. Calyces are fetched in containers of different sizes and prices are based on the size of the containers. There are export markets for Roselle products in countries such as the United States and Germany. Exported Roselle must meet some strict export standards including (1) moisture content (12%), (2) acidity, (3) residues, and (4) contaminations (e.g., waste and metals) (Mcclintock & Tahir, 2004). In Tanzania, a large market for the dried Roselle calyces is located at Dar es Salaam (ITM, 2008). Although there are poor local markets for Roselle calyces, there is a high potential for the development of processing industries; efforts have been directed at projects for juice production from cycles in both Kenya and Tanzania. In Tanzania, the Institute of Traditional Medicine of Muhimbili University of Health and Allied Science proposed to produce juice from a combination of Moringa leaves and Roselle cycles, such developments would improve the market potential of Roselle produce (ITM, 2008).

2.5.1.2 Marketing of seeds In the food industry, Roselle products are used as flavoring agents. Roasted Roselle seeds are used as food. Markets for Roselle seeds are seasonal with high demand during planting seasons and reduction in demands during other periods. Farmers may be required to store their seeds for extended periods of time to fetch good market prices; however, they may face challenges of pest damage. For example, extended storage of seeds in Niger for over a period of 7 months results in huge pest damage, therefore resulting in huge losses (Boureima, Moussa, & Lowenberg-DeBoer, 2015).

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2.6 Conclusion The growth and development of Roselle are strongly influenced by environmental conditions and genotype. Harvesting has a profound influence on the qualities of harvested Roselle, this ranges from harvesting time and standard postharvest processing procedures. Furthermore, postharvest handling has an influence on storage time and marketability of Roselle products. However, there are huge knowledge gaps on the appropriate procedures in harvesting and processing Roselle products. There is a potential market for Roselle products when marketing standards are met especially for export.

References Agarwal, J., & Dedhia, E. (2016). Sustainable development of Hibiscus sabdariffa-indigenous practices. International Journal of Science and Research (IJSR), 5(8), 190
194, Retrieved from. Available from https://www.ijsr.net/archive/v5i8/ART2016246.pdf. Ali, E. A., Kamarudin, K. M., Me, R. C., Sulaiman, R., & Alli, H. (2019). The development of SORREL systematic Roselle harvesting system for efficient transfer process. IOP Conference Series: Materials Science and Engineering, 697. Available from https://doi.org/10.1088/1757-899X/697/1/012029. Amoasah, B., Appiah, F., & Kumah, P. (2018). Effects of different drying methods on proximate composition of three accessions of Roselle (Hibiscus sabdariffa) calyces. International Journal of Plant & Soil Science, 21(1), 1
8. Available from https://doi. org/10.9734/ijpss/2018/38550. Bhat, R., Sridhar, K. R., & Yokotani, K. T. (2007). Effect of ionizing radiation on antinutritional features of velvet bean seeds (Mucuna pruriens). Food Chemistry, 103, 860
866. Boureima, S., Moussa, B., & Lowenberg-DeBoer, J. (2015). Analysis of the profitability of pic bags for the storage of Roselle grains (Hibiscus sabdariffa) in three regions in Niger. In: Working papers (August). Available from https://doi.org/10.22004/ag.econ.208450. Castro, N. E. Ade, Pinto, J. E. B. P., Cardoso, Mdas G., de Morais, A. R., Bertolucci, S. K. V., da Silva, F. G., & Delu´ Filho, N. (2004). Planting time for maximization of yield of vinegar plant calyx (Hibiscus sabdariffa L.). Cieˆncia e Agrotecnologia, 28(3), 542
551. Available from https://doi.org/10.1590/s1413-70542004000300009. Duke, J. A. (1983). Handbook of energy crops. West Lafayette, IN: Centre for New Crops and Plants Products, Purdue University. Gomaa, R. B. A., & Rashed, N. M. (2016). Post-harvest studies on reducing losses and maintaining quality of packaging Roselle calyxes. Journal of Sustainable Agricultural Sciences, 42(4), 68
86. Available from https://doi.org/10.21608/jsas.2016.3027. Haryati., Nisa, T. C., Julianti, E., & Blunden, G. (2018). Changes in size and weight of calyxes, weight, and moisture content of seeds of Roselle (Hibiscus sabdariffa L.) during development. Journal of Physics: Conference Series, 1116, 052027. Hussein, R. M., Shahein, Y., El Hakim, A. E., & Awad, H. M. (2010). Biochemical and molecular characterisation of three coloured types of Roselle (Hibiscus sabdariffa). Journal of American Science, 6(11), 726
733. Available from https://doi.org/10.7537/ marsjas061110.105.

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ITM (2008). Value chain analysis report for Morizella juice. ITM: Dar es Salaam. Kek, T. Z., Sakimin, S. Z., Nazrin, M., Juraimi, A. S., Alam, M. A., & Aslani, F. (2017). Regulation of fruit colour development, quality and storage life of Hibiscus sabdariffa L. As influenced by plant growth regulators. Bangladesh Journal of Botany, 46, 419
426. Mcclintock, N. C., & Tahir, E. (2004). Vegetables. In O. A. Grubben, & G. J. H. Denton (Eds.), Plant resources of tropical Africa 2 (pp. 321
326). Leiden/Wageningen: Backhuys Publishers/CTA. Mohamed, B. B., Sulaiman, A. A., & Dahab, A. A. (2012). Roselle (Hibiscus sabdariffa L.) in Sudan, cultivation and their uses. Bulletin of Environment, Pharmacology and Life Sciences, 1(6), 48
54. Mohamed, R., Fernadez, J., Pineda, M., & Aguilar, M. (2007). Roselle (Hibiscus sabdariffa) seed oil is a rich source of γ-tocopherol. Journal of Food Science, 72, 2007
2011. Morton, J. F. (1987). Roselle. In C. F. Dowling, Jr (Ed.), Fruits of warm climates (pp. 281
286). Greensboro, NC: Media Inc. Moussa, N., Ndeye, S. F., Joseph, B., & Mady, C. (2018). Stability of concentrated extracts of Hibiscus sabdarifa L. calyx during storage at different temperatures. African Journal of Food Science, 12(12), 347
352. Available from https://doi.org/10.5897/AJFS2018.1694. Nkafamiya, I. I., Atiku, J., Akinterinwa, A., & Fari, A. (2017). Effect of storage conditions on the degradation of Roselle (Hibiscus sabdariffa) seeds oil. International Journal of Biological and Chemical Sciences, 11(3), 1350. Available from https://doi.org/10.4314/ ijbcs.v11i3.34. Nnebue, O. M., Ogoke, I. J., Obilo, O. P., Agu, C. M., Ihejirika, G. O., & Ojiako, F. O. (2014). Estimation of planting dates for Roselle (Hibiscus sabdariffa L.) in the humid tropical environment of Owerri, South-eastern Nigeria. Agrosearch, 14(2), 168
178. Available from https://doi.org/10.4314/agrosh.v14i2.7. Nyarko, G., Bayor, H., Craigon, J., & Suleimana, I. A. (2006). The effect of container types, seed dressings and desiccants on the viability and vigour of Roselle (Hibiscus sabdariffa L. var sabdariffa) seeds. Pakistan Journal of Biological Sciences, 9(4), 593
597. Ochoa-Velasco, C. E., Salazar-Gonza´lez, C., Cid-Ortega, S., & Guerrero-Beltra´n, J. A. (2017). Antioxidant characteristics of extracts of Hibiscus sabdariffa calyces encapsulated with mesquite gum. Journal of Food Science and Technology, 54(7), 1747
1756. Available from https://doi.org/10.1007/s13197-017-2564-1. Plotto, A., Mazaud, F., Ro¨ttger, A., & Steffel, K. (2004). Hibiscus: Post-production management for improved market access organization. Rome, Italy: Food and Agriculture Organization of the United Nations (FAO), AGST. Saeed, I. E. (2010). Solar drying of Roselle (Hibiscus sabdariffa L.): Mathematical modelling, drying experiments, and effects of the drying conditions. Agricultural Engineering International: CIGR Journal, 12(3), 115
123. Shruthi, V. H., Ramachandra, C. T., Nidoni, U., Hiregoudar, S., Nagaraj Naik, N., & Kurubar, A. R. (2016). Roselle (Hibiscus sabdariffa L.) as a source of natural colour: A review. Plant Archives, 16, 515
522. Tindal, H. D. (1983). Vegetables in the tropics. London and Basingstoke: Macmillan Press Ltd.

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Effect of pests and diseases on Hibiscus sabdariffa quality

3

Gustav Komla Mahunu1, Maurice Tibiru Apaliya2 and Mildred Osei-Kwarteng3 1

Department of Food Science and Technology, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana 2 Department of Hotel Catering and Institutional Management, Cape Coast Technical University, Cape Coast, Ghana 3 Department of Horticulture, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana

Chapter Outline 3.1 Introduction ................................................................................................... 33 3.2 Pests of Roselle ............................................................................................. 34 3.2.1 Types of pests ...............................................................................34 3.2.2 Effect of pests on Roselle ..............................................................35 3.2.3 Prevention and control of pests ......................................................38 3.3 Diseases of Roselle ........................................................................................ 40 3.3.1 Types of diseases ..........................................................................40 3.3.2 Effect of diseases on Roselle ..........................................................41 3.3.3 Prevention and control of diseases ..................................................42 3.4 Conclusion ..................................................................................................... 43 References ............................................................................................................ 43

3.1 Introduction Pests and diseases present serious threats to crop production in tropical and subtropical regions of the world. Similar to the crops of Malvaceae family, Roselle is also highly susceptible to pests and diseases. All around the world, pests and diseases have emerged as a substantial limitation to the production of quality Roselle. Consistently over the years, the yield quality of the Roselle crop has been on the decline partly due to the effects of pests and diseases (Ambuja, Aswathanarayana, Govindappa, Naik, & Patil, 2017).

Roselle (Hibiscus sabdariffa). DOI: https://doi.org/10.1016/B978-0-12-822100-6.00004-5 © 2021 Elsevier Inc. All rights reserved.

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The intensive and extensive cultivation of high-yielding, fertilizer-responsive cultivars of Roselle have contributed to the problem of pests and nematodes. They have different feeding behaviors and feed on all parts of the plant, such as leaf defoliation. The degree of damage may vary between seasons, types, and among other essential factors. Pests attack on Roselle starts at the seedling stage and may cease prior to or persist in harvesting (Mahapatra, Mitra, Ramasubramanian, & Sinha, 2009). Although younger seedlings show greater susceptibility to these pests and their growth gets hampered, older plants may survive the attack but poor yield quality is irreversible. Findings indicate that various symptoms of Roselle diseases are attributed to fungi, bacteria, and virus infections, which can vary in appearance, incidence, intensity, and extent. Diseases have a destructive effect on Roselle plants, and severe cases often result in the death of the entire plant. Moreover, reports indicated that the manifestation and severity of the disease vary from one environment to the other. In the same way, different control measures of the disease, including manipulation of the host plant or its environment to a large extent, have helped to avoid some conditions. Generally, the population of pests and diseases, as well as their levels of invasion, are influenced by various factors such as soil moisture, rainfall, and plant growth stage. Moreover, the plants are most vulnerable to attacks in the seedling stage. Pests and diseases can cause a loss of up to 100% based on the crop and protection methods being used as, often, most of the attacks are not noticed until after a few months of field establishment. Given this, several investigations have proven that effective management of pests and diseases on crops can significantly contribute to food security by improving the availability and quality of food. Besides, new concerns have arisen due to the increasing importance of Roselle as high economic potential and as high export value crop. However, a comprehensive review of findings on the different aspects of Roselle, particularly pests and diseases, is seriously lacking. Undoubtedly, the availability of such information is essential for the provision of pests- and diseases-free conditions or significantly minimizing their occurrence; in certain instances, all these come to play as part of phytosanitary measures to fulfill export trade conditions. Hitherto, studies have been carried out to permit a better knowledge of the pests and diseases as well as their economic importance. This chapter reviews the occurrence and distribution of pests and diseases in Roselle, types of pests and diseases of Roselle, their effects on the growth and development of Roselle plants and, and, lastly, the preventive and control measures to reduce the effect of pests and diseases to reduce crop losses and improve productivity.

3.2 Pests of Roselle 3.2.1 Types of pests Roselle is affected by a variety of pests including stem borer, spiral borer Agrilus acutus Thunb, mealybug Maconellicoccus hirsutus, Phenacoccus solenopsis, and

3.2 Pests of Roselle

flea beetle Nisotra orbiculata (Motsch); abutilon moth, cotton bollworm, cotton Stainer and the cutworm, leaf miners, and the leafhoppers are the significant pests of Roselle (Hibiscus sabdariffa) (Plotto, Mazaud, Ro¨ttger, & Steffel, 2004). The pests attack different phonologies of Roselle plants, causing extensive economic damage. For instance, the pests attack the lamina of the young and mature leaves, leading to significant loss of the photosynthetic ability of the crop (Fasunwon & Banjo, 2010; Mahapatra et al., 2009). In a study by Abdel-Moniem, Abd El-Wahab, and Farag (2011), it was reported that 16 species of insect pests on Roselle belonged to 7 insect orders including Hemiptera (3 species), Homoptera (1 species), Lepidoptera (3 species), Neuroptera (1 species), Coleoptera (3 species), Diptera (1 species), and Hymenoptera (4 species). In addition, piercing sucking pests were the most predominant species; they include Empoasca spp., Spilostethus longulus Dallas, Nezara viridula L., and Oxycarenus hyalinipennis (Costa). An earlier report by Daramola (1984) outlined the insects and the corresponding damage they caused to Roselle plants (Table 3.1). Similarly, the predators that were observed include Polistes sp., Coccinella undecimpunctata, and Scymnus syriacus. Table 3.2 represents insect pests that perform predatory activities on Roselle. Honeybee (Apis mellifera L.) is mainly responsible for pollination on Roselle. Root-knot nematode is another economically important pest of Roselle. Among the various root-knot nematode species, Meloidogyne incognita, Meloidogyne javanica, and Meloidogyne arenaria mainly affect Roselle in the field (Adegbite, Agbaje, & Abidoye, 2008; Karssen & Moens, 2006; Minton & Adamson, 1979). However, a recent study by Ogunsola et al. (2018) identified four genera of root-knot nematodes in cultivated fields of Roselle: Meloidogyne spp., Helicotylenchus spp., Rotylenchulus spp., and Tylenchus spp. All were found in the roots and rhizosphere. The finding also showed that Meloidogyne had the highest population score (62.3%) on the roots with Tylenchus scoring the least (1.68%), and significant population density of Helicotylenchus spp. (50.1%) was recorded in rhizosphere but none was found on the roots.

3.2.2 Effect of pests on Roselle Mealybugs are polyphagous, sap-sucking pests with soft bodies, found worldwide. They have become economically important pests on varied agricultural and horticultural crops. Apart from causing direct damage to crops through their feeding activities, some mealybug species can act as conduits for spreading plant diseases. Crop losses attributed to destruction caused by mealybug alone has been estimated at up to 60%, and damage intensity was 4% between 40 and 65 days after planting (Satpathy, Gotyal, Ramasubramanian, & Selvaraj, 2013). Mealybugs will defoliate, puncture the tissues, consume the sap, and eventually weaken plants (Pena, Mohyuddin, & Wysoki, 1998). Some climatic conditions of precedence determine the general distribution, pattern, and intensity of pest infestation. A greater number of nonrainy days together with a slight rise in mean temperature by 1 C
2 C present favorable conditions for pest outbreak in the early stages of plant growth. Characteristically, tropical situation favors P. solenopsis by hastening the life cycle;

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Table 3.1 Insect pests and the parts they destroy of Roselle plant (Daramola, 1984). Order

Family

Genus

Species

Pest status

Part of plant destroyed

Coleoptera

Chrysomelidae Chrysomelidae Chrysomelidae Lagriidae Lagriidae Lagriidae Coccinellidae Telephoridae Lycidae Curculionidae Curculionidae Curculionidae Curculionidae Curculionidae Buprestidae Scarabaeidae Scarabaeidae Pyrgomorphidae Cercopidac Lygaeidae Lygaeidae Lygaeidae Pyrrhocoridae Pentatomidae Pentatomidae Coreidae Coreidae Coreidae Noctuidae Noctuidae

Podagrica Podagrica Syagrus Lagria Chrysolagrica Chrysolagria Epilaehna Silidius Lycus Lobotrachelus Nematocerus Alcidodes Baris sp. Apion sp. Pseudagrilus Pachnoda diplognatha Zonocerus Locris Oxycarenus Oxycarenus Lygaeus Dysdercus Nezara Drydocoris sp. Anoplocnemis Mirperus Clavigralla Earias Earias

sjostedti Jac. uniforma Jac. calcaratus F. villosa Fd ddc cuprina Thomas. nairobana Borch chrysomel ino F. bennensis Pic. semiamplexus Mur incalidus Boh. acerbus Fst. grassirostris Thorn NR NR sophorae F. cordata Dr. gagates F variegatus L. maculate F. hyalinipennis Costa gossypinus Dist. festivus Thull superstitiosus F. viridula L. NR -clirvipes F. torridus Wstw gibbosa Spin. biplaila Wlk insulana Bdr.

Major Major Major Major Major Major Minor Minor Minor NR NR NR Major Major Minor Minor NR Minor Minor Major Major Minor Minor Minor Minor Minor Minor Minor Major Major

Leaf Leaf Leaf Leaf Leaf Leaf Leaf Leaf Leaf Stem Stem Stem Fruits Fruits Stem Flower and fruit Flower and fruit Leaf and stem Leaf Fruit and seeds Fruit and seeds Fruit and seeds Fruit and seeds Fruit and seeds Fruit and seeds Fruit and seeds Fruit and seeds Fruit and seeds Fruit and seeds Fruit and seeds

Orthoptera: Hemiptera: -

Lepidoptcra: --

NR, not reported.

3.2 Pests of Roselle

Table 3.2 Insect pests that act as predators on Roselle. Order

Family

Genus

Species

Observed prey

Coleoptera

Coccinellidae Coccinellidae

Cheilomenes Cheilomenes

Aphids Aphids

Coccinellidae

Stictoleis

Coccinellidae

Cryptocephalus

Reduviidae Reduviidae

Rhinocoris Nagusta sp.

lunata F. vicina Muls mandata F. obesus Suff. bicolor F. NR

Hemiptera

Aphids Aphids Beetles, Oxycarenus spp. Dysdercus spp., Oxycarenllsspp.

NR, not reported.

FIGURE 3.1 Foliar damage by a beetle. Source: Photograph was taken from farmers field at Tampion Kukuo, Tamale, Northern Ghana.

thus its sensitivity to temperature and humidity fluctuations shortens the developmental period (Vennila et al., 2010). It was reported that warm and arid conditions during the dry season could trigger a mealybug outbreak on Roselle (Satpathy et al., 2013). Generally, it is can be concluded that the changing climatic conditions during the growing seasons influence the occurrence of most pests on Roselle. With this understanding, various preventive and control measures can be planned for the assurance of effective reduction of impact on crop productivity. It was also reported that the attack of flea beetles (Fig. 3.1) on Roselle begins with the seedling stage and ceases just before harvest. However, 7-day-old

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CHAPTER 3 Effect of pests and diseases on Hibiscus sabdariffa quality

seedlings are easily destroyed by flea beetles, and for that reason, resowing can be done to replace the lost young seedlings. Vegetables have many potentially destructive pests. Major pests including Podagrica sjostedti, Podagrica uniforma, Lagria villosa, Chrysolagria cuprina, Chrysolagria nairobana, and Syagrus calcaratus feed on the seedling and foliage. Also, the edible calyces and seeds were attacked by pests like Baris sp., Apion sp., and Earias spp. Their larvae feed on the inner part of the calyces and immature seeds and destroy them. Oxycarenus gossypinus and O. hyalinipennis also feed mostly on seeds. Baris and Apion damaged an estimated 10%
21% fruits harvested in mid-December and 48%
84% of late fruit harvests in mid-January. Percentage germination (10%) of insectinfested seeds was very poor, and uninfested seed germination was 83%. The root-knot nematode can cause 15%
20% damage in Roselle, although H. sabdariffa is relatively resistant (Laha & Pradhan, 1987; Mahapatra & Saha, 2008). The damage caused by the root-knot nematode enables the entry of opportunistic pathogens (such as bacteria, Ralstonia solanacearum, and fungi, Macrophomina phaseolina). The root-knot nematode creates injury on plant roots, causing substantial visible damage. Karssen and Moens (2006) reported that rootknot nematodes could change the physiology of the plant by producing specific enzymes that induce giant cell formation within the root at the feeding site.

3.2.3 Prevention and control of pests 3.2.3.1 Chemical control It was reported that farmers employ a wide array of pesticides (synthetic pyrethroids, organophosphates, phenyl pyrazoles, and neonicotinoids) without any substantial result (Satpathy et al., 2013). A possible reason for the poor effect of pesticides on pests was late application. Additionally, the resurgence of infestation beyond control can be caused by the remnants of the mealybug population on adjacent host plants (such as Parthenium hysterophorus, Abelmoschus esculentus, Solanum melongena, and Sesamum indicum). Another serious implication of repeated pesticides application is that activities of favorable natural enemies are suppressed (Satpathy et al., 2013). Though chemical control has been demonstrated to be very useful, it should be used very carefully as the possibility of pests developing resistance due to persistent pesticides application and the effect on the beneficial microbes have been reported. Foliage pests can be controlled effectively by spraying twice with carbaryl; spraying must be done at 2 weeks interval (first week apply 0.5 kg a.i./ha and the second week use 1.0 kg a.i./ha). However, to manage the effect of possible dosage hazard of carbaryl residues, 0.5 kg a.i./ha is recommended for the control of the leaf-eating insects on Roselle. This is because there were no significant differences between the effects of the two-treatment procedures. It was observed that fruit yield and seed viability of the treated Roselle plants were significantly higher compared to the untreated plants (Daramola, 1984). The carbaryl showed effective control of the Roselle insect pests that significantly improved the fruit yield.

3.2 Pests of Roselle

It was also reported that spiral borers could be effectively controlled with the application of carbofuran (1 kg/ha). Similarly, applying methyl demeton (2.5% @ 20 mL/plant) to wet the Roselle stem (at the height of 45 cm above the soil surface) can control spiral borers. However, some varieties of Roselle (RT 1) showed minimal susceptibility to A. acutus (Das et al., 2008).

3.2.3.2 Nonchemical control Organic farming appears more appropriate as it considers the important areas such as the sustainability of natural resources and the environment. Organic farming favors the maximum use of organic materials, including crop residue, animal residue, legumes, on- and off-farm wastages, growth regulators, and biopesticides. It discourages the use of synthetic agro-inputs to maintain soil productivity and fertility and to keep pests under control (Cid-Ortega & Guerrero-Beltra´n, 2015). A recent investigation showed that foliar spraying with ALKANZ 2000 and kaolin (Super Nano) formulations was able to decrease significantly the population of mealybug on Roselle (Hashem, EL-Hadidy, & Ali, 2017). These authors indicated that a special effect against mealybug insects was observed when EM bokashi (at 10 m3/Fed) was combined with ALKANZ 2000 or Super Nano. There was also an effective percentage reduction of leaf-miner insects infestation when EM bokashi was combined with Tracer 24% Sc or ALKANZ 2000. According to Abd ElAzim, Badawy, and Khater (2016), ALKANZ products comprise plant extracts with a variety of chemical components including many elements and active materials (such as tannins, carbohydrates, glycosides, flavonoids, and volatile and fixed oil). EM has been mentioned as a environment-friendly technology with the most significant effect in boosting plant growth, yield, and chemical constituents and at the same time suppressing wilt disease incidence and severity on Roselle (ElHadidy & El-Ati, 2014; Hashem et al., 2017; Widdiana & Higa, 1995). Ali (2016) describes Tracer 24% Sc as the first active ingredient that was proposed for a new class of insect control products, the Naturates. The positive effect of the Tracer (Naturates Spinosad) may be associated with the metabolites of the naturally occurring bacteria, Saccharopolyspora spinosa, which contain a mixture of Spinosyn A and Spinosyn D (Ali, 2016, 2017). This result agrees with a report on the effect of Azadiractin and Spinetoram treatment on the population density of the cotton aphid, Aphis gossypii that invade Roselle leaves (El-Zoghby, 2017). Kaolin actives as a pest repellant disrupt feeding and deter their oviposition (Alavo, Yarou, & Atachi, 2010). Kaolin is described as a white, nonabrasive, inert aluminosilicate mineral extensively used in various industrial applications including paints, cosmetics, and pharmaceuticals (Unruh, Knight, Upton, Glenn, & Puterka, 2000). Hydrophobic formulations of kaolin can effectively protect host plants against insect pests including lepidopterans, sucking insects, and mites. Through the foliar application, kaolin forms a layer on the surfaces of leaves, which then decreases transpiration rate, and retains more water in plant tissues for plant metabolism, photosynthetic rate, and increase the outward passage of photosynthates from the foliage to the fruits (Cantore, Pace, & Albrizio, 2009). The

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CHAPTER 3 Effect of pests and diseases on Hibiscus sabdariffa quality

plant coating with a hydrophobic particle film creates a barrier to suppress the attacks by making the plant visually or tactually unrecognizable as a host. As insects crawl upon the film, the particles get attached to the arthropod’s body, which severely reduced their movement, feeding, and other physical activities (Glenn, Puterka, Vanderzwet, Byers, & Feldhake, 1999). According to Gupta and Dikshit (2010), biopesticides are the eco-friendly alternative to chemical pesticides, encompassing a broad range of microbial pesticides, biochemicals derived from microorganisms, and other natural sources. As one of the natural products, biopesticides are involved directly in controlling insects, plant pathogens, weeds, and nematodes in the field or indirectly by improving plant health (Koivunen, Duke, Coats, & Beck, 2013). The overreliance on chemical pesticides for conventional agricultural production over time has led to serious challenges. Chemical pesticides used haphazardly have contributed to health deterioration, environmental degradation, and unsustainable systems. Therefore the use of organic materials and compounds of natural origins is the way forward for sustainable agricultural production systems (Bhanti & Taneja, 2007). It has been recognized that preventing the occurrence of root-knot nematode is the main approach to effective control; this is because the cost of controlling advanced heavy infestation is a very high or difficult task to attain. Basic cultural practices including removal of stubbles, weed control, thinning of plants, and 2-year crop rotation with rice are methods to minimize root-knot nematode field colonization (Mahapatra et al., 2009). Intercropping or mixed cropping has also been found as a suitable alternative to reduce the pest population. All these approaches have shown comparable advantages to the chemical method without compromising the positive effect of beneficial microorganisms. A number of varietal screening studies on Roselle types identified just a few as having resistance to specific insect pests. In an earlier report, Malati and Namita (1988) identified five kinds of Roselle belonging to red-bristled types showing different resistant levels: one possessing a good resistant reaction while the other four had moderately resistant characteristics against Phytophthora parasitica in the field.

3.3 Diseases of Roselle 3.3.1 Types of diseases Fungi are major disease-causing microorganisms in Roselle. The fungal species that cause diseases in Roselle include Aecidium garckeanum, A. hibiscisurattense, Alternaria macrospora, Cercospora abelmoschii, C. malaysensis, Corynespora cassiicola, Cylindrocladium scoparium, Diplodia hibiscina, Fusarium decemcellulare, F. sarcochroum, F. solani, F. vasinfectum, Guignardia hibisci sabdariffa, Irenopsis molleriana, Leveillula Taurica, Microsphaera euphorbiae, Phoma sabdariffae, Phymatotrichum omnivorum, P. parasitica, Phytophthora terretris,

3.3 Diseases of Roselle

Pythium perniciosum, Rhizoctonia solani, Sclerotinia fuckeliana, S. sclerotiorum, and Sclerotium rolfsii. P. parasitica var. sabdariffae is the causal organism of root and stem rot, soft rot or collar rot is caused by S. rolfsii and yellow vein mosaic is caused by geminivirus (Mahapatra et al., 2009). Other authors also stated that among the diverse organisms causing stem and root rot on Roselle, the following fungal species are the most outstanding ones: P. parasitica, R. solani, M. phaseolina, S. rolfsii, Phymatotrichopsis omnivora, Fusarium oxysporum, F. semitectum, F. solani, and F. equiseti (Amusa, Adegbite, & Oladapo, 2005; Hassan, Shimizu, & Hyakumachi, 2014; Ploetz, Palmateer, Geiser, & Juba, 2007). Ortega-Acosta et al. (2015), in their study, confirmed that P. parasitica was the most commonly occurring species, whereas F. oxysporum was observed to be the low occurring pathogen of Roselle. Other studies on molecular characterization identified two distinct whiteflytransmitted monopartite begomovirus complexes associated with the yellow vein mosaic disease. The two are the yellow vein mosaic virus and yellow vein mosaic Bahraich virus of Roselle. Two other species of the beta satellite are the cotton leaf curls Multan beta satellite and Ludwigia leaf distortion beta satellite (Roy et al., 2009). The complexes of begomovirus were observed in different combinations across many Roselle cultivation regions in India. Arif et al. (2018) recently confirmed that cotton leaf curl Multan virus (CLCuMuV-CLCuMuB) was first identified in Roselle. Also, Roselle is the fourth seed-propagated plant to suffer from this disease in China. In another interesting study, Eslaminejad and Zakaria (2011) fungal pathogens associated with Roselle diseases were identified and characterized according to their morphological and cultural characteristics to determine the pathogenicity. These authors found that four fungi (Phoma exigua, Fusarium nygamai, F. camptoceras, and R. solani) were present in the samples of Roselle seedlings. P. exigua was in 45% of the samples which is more than F. nygamai (25%), R. solani (19%), and F. camptoceras (11%). Lastly, the pathogenic test on these fungi also established different levels of severity.

3.3.2 Effect of diseases on Roselle Foot and stem rot are the more severe diseases in Roselle, and due to severe infection, more than 40% crop loss was recorded (De & Mandal, 2007; Ghosh, 1983). It was observed that the diseased plant produces secondary roots above the rotten portion of the stem, as a way of struggling to survive the severe infections. In addition, the appearance of “blackleg” includes basal necrosis, general wilting, leaves acquiring a yellow color, and finally death of the plant. These were found as one of the main areas that limit plant health for Roselle production (OrtegaAcosta et al., 2015). Collar rot occurs in Roselle, although the incidences are not yet alarming (De & Mandal, 2007). However, various levels of yield losses due to the effect of yellow

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CHAPTER 3 Effect of pests and diseases on Hibiscus sabdariffa quality

vein mosaic have been recorded in Roselle. Fiber yield loss was approximately 12.8%
17.5%, and 18.9%
23.8% for seed yield loss have been reported. The Viruliferous whitefly has also been observed to carry out an effective transmission of yellow vein mosaic at 78%
85% transmission efficiency where three of these whiteflies per Roselle plant can adequately transmit the yellow vein mosaic disease (YVMD) on the plant (Roy et al., 2007). Studies on the interaction effect of whitefly vector populations, climatic conditions, and disease spread were conducted, which cited the impact of favorable weather as the main factor that enabled whitefly populations to spread YVMD (Chatterjee & Ghosh, 2015; Roy et al., 2007). The disease is endemic, and the rate of spread is faster. The first observation of yellow vein mosaic disease in Roselle was made in Andhra Pradesh and then reported from West Bengal (Chatterjee, Roy, Padmalatha, Malathi, & Ghosh, 2005; Ghosh, 2009). Eight cultivars of Roselle were tested, and all were very susceptible to YVMD. Yellow vein mosaic disease
infected plants will lose about 70% of the fiber yield when inoculation occurs at an early stage of growth (Roy et al., 2009). Studies showed that continuous cultivation of infected fields could lead to more than 40% of Roselle crops being blight disease infected. The leaf blight can cause more than 20% loss of leaf yield and more than 34% loss of edible leaf. The study also recorded that average biological and marketable yield losses of calyces were 35% and 38%, respectively, whereas the biological and commercial yield losses of seed represent 30% and 32% (Amusa, 2004).

3.3.3 Prevention and control of diseases 3.3.3.1 Chemical It was reported that treatment of Roselle seeds with copper oxychloride was more effective against P. parasitica (De & Mandal, 2007) and could reduce disease incidence at 30 days after sowing by 50.3% and at maturity by 45.5%. Also, spraying with copper oxychloride at a concentration of 5
7 g/L of water can control collar rot; however, when the chemical is directed to the base of the Roselle plants, it provides effective control of the disease (Mahapatra et al., 2009).

3.3.3.2 Nonchemical control of Roselle diseases Use of the appropriate Roselle seeds for planting can prevent infection, as some cultivated varieties have exhibited resistance to diseases caused by P. parasitica. Screening of cultivated varieties of Roselle (“AMV 1,” “RT 1”, and “AP 481”) found them to be moderately resistant to P. parasitica (Malati & Namita, 2000). In other words, varietal selection of Roselle within a growing area can be relied on to control the pathogen. Sowing time is another crucial factor that determines the resistance or susceptibility of crops to pests and diseases. It has a direct or an indirect implication on the agronomic and yield characters of crops. Therefore the choice of the

References

appropriate sowing time in a particular season is a critical decision that can significantly enhance crop yield. It was stated that the occurrence of viral diseases on crop fields under sub-Saharan tropical conditions might not be noticeable at the best sowing time. A study report Kareem, Oduwaye, Olanipekun, Adeniyan, and Oyedele (2017) concluded that June is the most suitable sowing time for Roselle, when relatively low virus incidence (such as YVMD) occurs coupled with attainment of the highest plant height and bast fiber yield. However, due to the changing climatic conditions, the sowing time may not be fixed over the years, which means that dependence on sowing time ought to be a continuous process for the improvement of crop yield.

3.4 Conclusion In conclusion, the poor management of Roselle plants thriving can lead to reduced light penetration, reduced plant productivity, low color development of fruit, declined quality of fruit and profit, and eventually affects pest and disease control negatively. Future studies should be conducted on storage pests and diseases of Roselle due to limited information available. For instance, the triple bagging technology (TBT) is offering a promising alternative that can be manipulated to favor the postharvest storage of many crops, including Roselle. However, there is a need to validate the efficiency of TBT against the diversity of pests of Roselle, which possess different behavior and evolution. Lastly, the implementing strategies against pests of Roselle must be comprehensive and integrative.

References Abd El-Azim, W., Badawy, M., & Khater, R. (2016). Effect of Mycorrhiza and phosphate dissolving bacteria and licorice extract on growth and oil productivity of Foeniculum vulgare, mill. plant under Baluza in North Sinai conditions. Middle East Journal of Applied Sciences, 6(4), 990
1002. Abdel-Moniem, A., Abd El-Wahab, T., & Farag, N. (2011). Prevailing insects in Roselle plants, Hibiscus sabdariffa L., and their efficiency on pollination. Archives of Phytopathology and Plant Protection, 44(3), 242
252. Adegbite, A., Agbaje, G., & Abidoye, J. (2008). Assessment of yield loss of Roselle (Hibiscus Sabdariffa L.) due to root-knot nematode, Meloidogyne incognita under field conditions. Journal of Plant Protection Research, 48(3), 267
273. Alavo, T., Yarou, B., & Atachi, P. (2010). Field effects of kaolin particle film formulation against major cotton lepidopteran pests in North Benin, West Africa. International Journal of Pest Management, 56(4), 287
290. Ali, E. (2016). Effectiveness of particle film technology and copper products in the control of olive fruit fly. Journal of Plant Protection and Pathology, 7(7), 439
444.

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Ali, E. (2017). Integrated control of some pests infesting pomegranate trees in Northern Western coast. Egyptian Academic Journal of Biological Sciences, 9(1), 59
72. Ambuja, H., Aswathanarayana, D., Govindappa, M., Naik, M., & Patil, M. (2017). Survey and biological characterization of leaf curl disease of mesta (Hibiscus sabdariffa L.). Journal of Pharmacognosy and Phytochemistry, 6(6), 1949
1954. Amusa, N. (2004). Foliar blight of Roselle and its effect on yield in tropical forest region of Southwestern Nigeria. Mycopathologia, 157(3), 333
338. Amusa, N., Adegbite, A., & Oladapo, M. (2005). Vascular wilt of Roselle (Hibiscus sabdariffa L. var. sabdariffa) in the humid forest region of South-western Nigeria. Plant Pathology Journal, 4(2), 122
125. Arif, M., Lin, W., Lin, L., Islam, W., Jie, Z., He, Z., & Wu, Z. (2018). Cotton leaf curl Multan virus infecting Hibiscus sabdariffa in China. Canadian Journal of Plant Pathology, 40(1), 128
131. Bhanti, M., & Taneja, A. (2007). Contamination of vegetables of different seasons with organophosphorous pesticides and related health risk assessment in northern India. Chemosphere, 69(1), 63
68. Cantore, V., Pace, B., & Albrizio, R. (2009). Kaolin-based particle film technology affects tomato physiology, yield and quality. Environmental and Experimental Botany, 66(2), 279
288. Chatterjee, A., & Ghosh, S. K. (2015). Yellow vein mosaic disease: A new threat to mesta (Hibiscus sp.) cultivation. Recent advances in the diagnosis and management of plant diseases (pp. 145
161). Springer. Chatterjee, A., Roy, A., Padmalatha, K., Malathi, V., & Ghosh, S. (2005). Occurrence of a begomovirus with yellow vein mosaic disease of mesta (Hibiscus cannabinus and Hibiscus sabdariffa). Australasian Plant Pathology, 34(4), 609
610. Cid-Ortega, S., & Guerrero-Beltra´n, J. (2015). Roselle calyces (Hibiscus sabdariffa), an alternative to the food and beverages industries: A review. Journal of Food Science and Technology, 52(11), 6859
6869. Daramola, A. M. (1984). The insect pest complex of the edible Roselle Hibiscus sabdariffa var. sabdariffa L and the damage they cause in Southwestern Nigeria. Nigerian Journal of Entomology, 5, 62
69. Das, S., Roy, A., Ghosh, R., Paul, S., Acharyya, S., & Ghosh, S. K. (2008). Sequence variability and phylogenetic relationship of betasatellite isolates associated with yellow vein mosaic disease of mesta in India. Virus Genes, 37(3), 414
424. De, R., & Mandal, R. (2007). Effect of seed treatment with fungicides on foot and stem rot disease caused by Phytophthora parasitica var. sabdariffae in Hibiscus sabdariffa. Journal of Interacademicia, 11(2), 161
165. El-Hadidy, A., & El-Ati, A. (2014). Efficiency of effective microorganisms (EM) to induce resistance against chocolate spot disease and enhance productivity of faba bean under reclaimed soil conditions. Egyptian Journal of Phytopathology, 42(1), 117
142. El-Zoghby, I. (2017). Studies on the impact of successive sprays with certain insecticides on whitefly and aphids infesting Roselle plants and its yield in Aswan Governorate, Egypt. Middle East Journal of Applied Sciences, 7(1), 162
167. Eslaminejad, T., & Zakaria, M. (2011). Morphological characteristics and pathogenicity of fungi associated with Roselle (Hibiscus sabdariffa) diseases in Penang, Malaysia. Microbial Pathogenesis, 51(5), 325
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771. Gupta, S., & Dikshit, A. (2010). Biopesticides: An ecofriendly approach for pest control. Journal of Biopesticides, 3, 186. Hashem, H., EL-Hadidy, A. E., & Ali, E. (2017). Impact of some safe agricultural treatments on insect pests, vascular wilt disease management and Roselle (Hibiscus sabdariffa L.) productivity under Siwa Oasis conditions. International Journal of Environment, 6(4), 139
162. Hassan, N., Shimizu, M., & Hyakumachi, M. (2014). Occurrence of root rot and vascular wilt diseases in Roselle (Hibiscus sabdariffa L.) in Upper Egypt. Mycobiology, 42(1), 66
72. Kareem, K., Oduwaye, O., Olanipekun, S., Adeniyan, N., & Oyedele, A. (2017). Yellow vein mosaic disease in kenaf (Hibiscus cannabinus L.) under different sowing dates in two agroecologies. Nigerian Journal of Biotechnology, 33(1), 83
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4). ACS Publications. Laha, S., & Pradhan, S. (1987). Susceptibility of bast fibre crops to Meloidogyne incognita. Nematologia Mediterranea, 15, 163
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37). Barrackpore, Kolkata: CRIJAF. Mahapatra, B., Mitra, S., Ramasubramanian, T., & Sinha, M. (2009). Research on jute (Corchorus olitorius and C. capsularis) and kenaf (Hibiscus cannabinus and H. sabdariffa): Present status and future perspective. Indian Journal of Agricultural Sciences, 79 (12), 951
967. Malati, M., & Namita, M. (1988). Field evaluation of Roselle germplasm for resistance against foot and stem rot caused by Phytophthora parasitica Dast. International Journal of Tropical Plant Diseases, 6(1), 73
76. Malati, M., & Namita, M. (2000). Comparative yield performance of some resistant and susceptible genotypes and the check varieties of Roselle (Hibiscus sabdariffa L.). Journal of Mycopathological Research, 38(1), 51
52. Minton, N., & Adamson, W. (1979). Control of Meloidogyne javanica and M. arenaria on kenaf and Roselle with genetic resistance and nematicides. Journal of Nematology, 11(1), 37. Ogunsola, K., Ogunfunmilayo, A., Solomon, S., Oluitan, J., Kazeem, S., Folorunso, D., & Ibrahim, S. (2018). Occurrence and population distribution of plant parasitic nematodes associated with Roselle (Hibiscus sabdariffa L.) in northern Nigeria. Agro-Science, 17(2), 18
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Ortega-Acosta, S., Herna´ndez-Morales, J., Sandoval-Islas, J., Ayala-Escobar, V., SotoRojas, L., & Alejo-Jaimes, A. (2015). Distribution and frequency of organisms associated to disease "black leg" of Roselle (Hibiscus sabdariffa L.) in Guerrero, Me´xico. Revista Mexicana de Fitopatolog´ıa, 33(2), 173
194. Pena, J., Mohyuddin, A., & Wysoki, M. (1998). A review of the pest management situation in mango agroecosystems. Phytoparasitica, 26(2), 129. Ploetz, R., Palmateer, A., Geiser, D., & Juba, J. (2007). First report of Fusarium wilt caused by Fusarium oxysporum on Roselle in the United States. Plant Disease, 91(5), 639. Plotto, A., Mazaud, F., Ro¨ttger, A., & Steffel, K. (2004). Hibiscus: Post-production management for improved market access organisation. Food and Agriculture Organization of the United Nations (FAO). AGST. Roy, A., Acharyya, S., Das, S., Ghosh, R., Paul, S., Srivastava, R. K., & Ghosh, S. K. (2009). Distribution, epidemiology and molecular variability of the begomovirus complexes associated with yellow vein mosaic disease of mesta in India. Virus Research, 141(2), 237
246. Roy, A., Ghosh, R., Paul, S., Das, S., Palit, P., Acharyya, S., Ghosh, S. (2007). Characterization of a monopartite recombinant begomovirus and satellite DNA beta associated with yellow vein mosaic disease of mesta crop in India. Paper presented at the Proceedings of 10th plant virus epidemiology symposium, ICRISAT, India. Satpathy, S., Gotyal, B., Ramasubramanian, T., & Selvaraj, K. (2013). Mealybug, Phenacoccus solenopsis Tinsley infestation on jute (Corchorus olitorius) and mesta (Hibiscus sabdariffa). Insect Environment, 19(3), 187
188. Unruh, T., Knight, A., Upton, J., Glenn, D., & Puterka, G. (2000). Particle films for suppression of the codling moth (Lepidoptera: Tortricidae) in apple and pear orchards. Journal of Economic Entomology, 93(3), 737
743. Vennila, S., Deshmukh, A., Pinjarkar, D., Agarwal, M., Ramamurthy, V., Joshi, S., & Bambawale, O. (2010). Biology of the mealybug, Phenacoccus solenopsis on cotton in the laboratory. Journal of Insect Science, 10(1), 115. Widdiana, G., & Higa, T. (1995). Effect of EM on the production of vegetable crops in Indonesia. Paper presented at the 4th Proceedings International Conference of Kysei Nature Farmingheld in Paris, France.

CHAPTER

Measurement and maintenance of Hibiscus sabdariffa quality

4

Joseph Patrick Gweyi-Onyango1, Mildred Osei-Kwarteng2 and Gustav Komla Mahunu3 1

Department of Agricultural Science and Technology, Kenyatta University, Nairobi, Kenya Department of Horticulture, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana 3 Department of Food Science and Technology, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana

2

Chapter Outline 4.1 Background .................................................................................................... 47 4.2 Quality and quality maintenance of the Hibiscus sabdariffa ............................... 49 4.2.1 Anthocyanins as an essential component of Hibiscus sabdariffa quality .........................................................................................50 4.2.2 Microbial and antimicrobial activities and Hibiscus sabdariffa quality 51 4.2.3 Influence of plant growth, age, and harvesting on Hibiscus sabdariffa quality .........................................................................................52 4.2.4 Antinutritional components and the prevailing factors on Hibiscus sabdarrifa quality ..........................................................................54 4.3 Maintenance of quality of secondary metabolites of Hibiscus sabdariffa ............ 55 4.4 Methods of measurements of bioactive compounds in Hibiscus sabdariffa ......... 57 References ............................................................................................................ 59

4.1 Background Hibiscus sabdariffa (Roselle; Malvaceae) is well-known in the Middle East states (Abu-Tarboush, Ahmed, & Al Kahtani, 1997). Besides the Middle East, it also grows in other tropical nations in South East Asia; Malaysia, Indonesia, Philippines, and Thailand (Chewonarin et al., 1999) and West Africa; Senegal, Mali, Burkina Faso, Ghana, Benin, Nigeria, and Niger (Achoribo, Achel, Gibrilla, Adaboro, & Palm, 2012). The green variety of this crop is very rich in vitamin C, riboflavin, β-carotene, and key mineral elements (Babalola, 2000). The petals Roselle (Hibiscus sabdariffa). DOI: https://doi.org/10.1016/B978-0-12-822100-6.00008-2 © 2021 Elsevier Inc. All rights reserved.

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have a wide range of usage, including herbal drinks, warm, and cold beverages, and some products of jams and jellies (Tsai, Shun-Fu Tseng, Chang, Lin, & Teng, 2002). Their unique flavor and brilliant red color make them have high food-product value (Tsai et al., 2002). Roselle is either an annual or perennial herb with edible flowers (Ram´ırez-Rodrigues, Balaban, Marshal, & Rouseff, 2011). The swollen calyces, with different shades of red to blue and cup-like shape, are the parts that are of commercial interest (De Castro et al., 2004). The plant is rich in dietary fiber and possesses high amounts of bioactive compounds or phytochemicals including anthocyanins and carotenoids (Ozlem, 2011). It also contains a wide range of nonstarchy polysaccharides (Borra´s-Linares et al., 2015). The phytochemicals such as phenolic compounds extracted from hibiscuses are used in diverse ways including antioxidants and hyperlipidemia and are also effective against low-density lipoprotein (Sa´yago-Ayerdi, Arranz, Serrano, & Gon˜i, 2007), although they are quite sensitive to oxygen, extreme temperatures, and ultraviolet (UV) light (Tolun, Altintas, & Artik, 2016). The calyces are endowed with high quantities of anthocyanins that are used in the manufacture of flavored jams, jellies, and beverages (Hirunpanich et al., 2006). Furthermore, the anthocyanins have coloring properties that make them suitable for the food industry as they provide appealing hues and aroma to foods (Reyes and Cisneros-Zevallos, 2007). It cannot be overemphasized that many edible parts, such as hibiscus calyces, are significant sources of anthocyanins, which are water-soluble pigments in plants (Wong, Yusof, Ghazali, & Man, 2002). Hibiscus encompasses anthocyanins such as cyanidin, cyanidin-3-sambubioside, delphinidin, delphinidin-3-sambubioside, and malvidin (Preciado-Saldan˜a et al., 2019). Hibiscus calyx has been used to produce infusions or teas and natural dyes (Piovesana & Noren˜a, 2019). Some exciting reports reveal that Roselle is used in brewing teas to treat several disorders; among them are diabetes, high blood pressure, constipation, cancer, heart disease, urinary tract infections, and hepatic disorders that have been documented in traditional medicine (Ferna´ndez-Arroyo, Herranz-Lo´pez, & Beltra´n-Debo´n, 2012; McKay, Chen, Saltzman, & Blumberg, 2010). Roselle tea has a unique trait that relates to its appearance, aroma, flavor, and color, but aroma is the most crucial determinant of tea quality to meet consumer preferences (Monteiro, Costa, Tomlins, & Pintado, 2019). Indeed, very tasty and highly acceptable delicious different tea blends have been produced from H. sabdariffa (L), Cymbopogon citratus, and Citrus limon mixture, as stated by Ogbonna, Makut, Gyar, Ogbonna, and Wogu (2010). In their study, the tea tasters’ panel overwhelmingly placed hibiscus tea in the category and range of other know commercial teas. The formulated tea appeared attractive and had no significant difference between it and the control (Camellia sinensis; Ezearigo, Adeniji, & Ayoade, 2014) in color and taste (Keser et al., 2020). Also, foodproduct value is depicted by brilliant red color and its unique flavor. The pigments in anthocyanin for color are attributed to many other colors in various foods (Tsai & Ou, 1996).

4.2 Quality and quality maintenance of the Hibiscus sabdariffa

Recently, processors have turned to the use of colorants derived from natural products as an alternative in response to the increased consumer demands for these natural products instead of synthetic counterparts. In this context, the availability of natural pigments, including the sources, the extraction and the concentration processes, and the colorants stability, should be considered in the production of these ingredients as observed by Patil, Madhusudhan, Ravindraand, and Raghavarao (2009). Generally, color intensity is a significant index in calyx utility for beverage drinks. Concentrations have often been analyzed in the spectrophotometric assay of extracts, with findings portraying significant variances between varieties. In differentiating varieties color intensity, sourness to taste and overall acceptability to drink, sensory, and chemical analysis is carried out (Patil et al., 2009).

4.2 Quality and quality maintenance of the Hibiscus sabdariffa The cultivation of Roselle calyces with good characteristics in emerging regions is essential in value addition and high revenue generation for rural livelihoods. Nonetheless, evidence on actual quality assessment and improvement is scanty in the literature particularly the number of active ingredients and cleanliness are always missing in commercial standards. Juliani et al. (2009) have in the past attempted to provide information on chemistry and quality of this plant with the aim to develop natural products. Aroma is critical in determining the overall tea quality and consumers’ tastes and preferences, and it is affected by diverse factors like drying, heating, and brewing (Zannou, Kelebek, & Selli, 2020). Aromatic compounds that contain molecular weights of less than 300 Da vaporize readily at room temperature (Keser et al., 2020). Hence, in Roselle, the aroma composition is dependent on processing factors and enzymatic reactions and lipid oxidation (Zannou et al., 2020). During the processing of other foods, the volatile components change and degrade and may lead to inferior quality final food product (Zannou et al., 2020) and consequently affect the acceptance of the product by consumers (Duan & Barringer, 2012). Therefore further considerations such as processing, storage, and management should be explored to maintain quality and acceptance of the Roselle products. Management practices determine the final product of most crop plants, including Roselle. Generally, the quality of natural Roselle products and many crops depends on production and postharvest handling techniques and other factors like genetic selection and system processing that have a great impact on the quality of the product, particularly the bioactive components (Nedovi´c et al., 2016). The quantity of bioactive compounds influences quality, thus, optimizing their content during production and processing for commerce and making products with improved functional properties (Juliani et al., 2009). Therefore the occurrence of

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simple analytical procedures for determining polyphenolic compounds is essential in product quality assessment (Juliani et al., 2009). Production and assurance of hygienic products (Juliani et al., 2009) are vital for the health of the consumer (Elhassan, Ahmmed, & Sirag, 2014). The general purpose is to increase active or functional elements in the tea or beverage. A functional drink can be defined as one that exerts beneficial effects that result in a decreased risk of disease or has undergone technological modifications to provide a benefit (Corbo, Bevilacqua, Petruzzi, Casanova, & Sinigaglia, 2014). According to this definition, a beverage prepared from H. sabdariffa can be considered functional when technological means are used to maximize its anthocyanin content and antioxidant activity to minimize the consumers’ risk of disease increased anthocyanin intake. There are many ways of improving functional beverages. Dry Roselle extracts can produce a stable powder that has a long storage life. During the drying process, a carrier agent is needed to capture active agents within the carrier material (Nedovic, Ana, Verica, Steva, & Branko, 2011), to improve bioactive delivery and must be food grade, recyclable, and able to form a wall or barrier between the internal phase and its surroundings (Ram´ırez, Giraldo, & Orrego, 2015). The most common wall materials are maltodextrin, gum arabic, and modified starch (Ram´ırez et al., 2015). These carbohydrate media coat the active compound in a transparent phase and protect the active compound from higher temperature, collapse, and enzymatic or chemical changes, such as oxidation (Ram´ırez et al., 2015).

4.2.1 Anthocyanins as an essential component of Hibiscus sabdariffa quality Anthocyanins are highly bio-reactive compounds, and their intake from H. sabdariffa infusions has shown to elicit positive impacts on the consumers’ health. A metaanalysis of data concluded that H. sabdariffa could significantly decrease systolic and diastolic blood pressure of hypertensive people (Serban, Sahebkar, Ursoniu, Andrica, & Banach, 2015). Others have demonstrated an antitumor effect against different cancerous cells of human origin, and their effects on a rat model of chronic kidney disease were similar to pharmacological antihypertensive (Ali et al., 2017; Chiu, Chen, Chou, & Lin, 2015). The environmental conditions during Roselle production affect the primary polyphenols (including anthocyanins) present in the calyces. Depending on the production site, the amount of anthocyanins can double in some varieties (Juliani et al., 2009). The increase in the anthocyanin content in the calyces could improve the nutraceutical quality and increase the product’s value. Currently, in Roselle marketing (see Chapter 2: Harvesting, Storage, Postharvest Management, and Marketing of Hibiscus sabdariffa), the essential quality criteria are the acidity of the extracts and the anthocyanin content; the latter is related to the number of extractions that can be made from the calyces (Moreno, Birstein, Mayer, Silva, & Soto, 2008).

4.2 Quality and quality maintenance of the Hibiscus sabdariffa

Generally, anthocyanins are pigments produced by secondary metabolism; and their concentration can increase in plants in response to oxidative stress caused by several factors, including exposure to high concentrations of metals (particularly heavy metals), drought, salinity, and UBV (UV, blue, and visual) tend to increase polyphenol contents and profiles. Herna´ndez, Alegre, van Breusegem, and Munne´-Bosch (2009) argued that there is significant activation of some essential plant genes under these conditions, which consequently modulate the biosynthesis of flavonoids such as anthocyanins, with resultant higher levels of these phytochemicals when exposed to varying levels of stress. The inductive synthesis of anthocyanins due to external cues is the outcome of gene activation, which takes place and coincidentally enhances the increase in anthocyanins or antioxidant response of the plant to maintain the physiological status of tissues that are affected by stress either directly or indirectly (LingPeng, Xin-Jiao, & Hai-Hu, 2012).

4.2.2 Microbial and antimicrobial activities and Hibiscus sabdariffa quality H. sabdariffa has been used in West African countries such as Togo, Ghana, and Nigeria in making “zobo” drink. Zobo is derived from “zoborodo” a Hausa name for H. sabdariffa (Aliyu, 2000). Available information shows that this is exclusively a nonalcoholic drink that is popular with the local folks where it is mostly served at social gatherings (Aliyu, 2000; Nwachukwu, Onovo, & Ezeama, 2007). In the recent past, there has been an increase in zobo for no apparent reason. However, speculations point to a few reasons: (1) its low price, (2) nutritional, and (3) medicinal attributes related to this drink in particular and hibiscus-related products in general. Despite all the right attributes related to consumption, there are critical challenges that need to be addressed to allow mass production of zobo drink, due to the rapid deterioration of the drink following production if not refrigerated (Nwachukwu et al., 2007). The possible argument for this quick deterioration of the drink is the presence of some microorganisms that cause food spoilage (Risiquat, 2013). The candidate microbes include Escherichia coli, Bacillus spp., Staphylococcus aureus, Streptococcus faecalis, Proteus spp., Enterobacter spp., Klebsiella spp. (Risiquat, 2013), Micrococcus spp., Aspergillus spp., Penicillium spp., Fusarium spp., Rhizopus spp., and Mucor spp. (Medina et al., 2006). Microbe infections are silently but critically becoming a concern and may hamper the upward demand for zobo as food safety is a crucial requirement. Food safety is an essential concern for both consumers and food producers (Witkowska, Hickey, Alonso-Gomez, & Wilkinson, 2013). However, not all food microbes are necessarily detrimental as their growth may impart a pleasant taste and texture (Salami & Afolayan, 2020). Unfortunately, most microorganisms such as S. aureus, Pseudomonas aeruginosa, E. coli, Vibrio cholerae, etc. are associated with massive spoilage, and food contamination also transfers a wide range of disease conditions in food (including zobo; Bukar, Uba, & Oyeyi, 2010).

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Packaging material, particularly bottles and polythene bags used by retailers, are generally not ideal for the drink. There are significant entry points for microbes that contaminate the zobo drink. Besides packaging, key sources of contamination are the dried calyces as they harbor decay organisms such as Penicillium and Aspergillus sp. (Ezearigo et al., 2014). In general, many retailers do not observe hygiene in the preparation of the drink and fail to properly subject the drink to adequate heating to decrease the microbial load during the preparation and also when in storage (Ezearigo et al., 2014). On the other hand, to the relief of producers and consumers, reports show that extracts of ginger, garlic, cinnamon, and nutmeg are used in foods as natural antimicrobials (Ezearigo et al., 2014). These natural antimicrobials can extend the shelf life of zobo from the traditional 24
48 hours to about a week (Witkowska et al., 2013). Furthermore, the extracts of traditional natural spices (including Roselle) are known to have a broad spectrum antibacterial activity including effects on Escherichia, Salmonella, Staphylococcus, Streptococcus, Klebsiella, Proteus, Clostridium, Mycobacterium, and Helicobacter species (Ayoade et al., 2012). Research has lately shown that Roselle calyces have a potent antimicrobial consequence on pathogenic bacteria (Cid-Ortega & Guerrero-Beltra´n, 2015). It has been reported that aqueous extracts of Roselle calyces were used to sanitize romaine lettuce and alfalfa sprouts infected with E. coli, Salmonella, and Listeria monocytogenes (Gomez-Aldapa et al., 2018) as well as to effectively cleanse organic iceberg lettuce and spinach contaminated with Salmonella (GomezAldapa et al., 2018). Further, Gomez-Aldapa et al. (2018) reported that four different extracts (in water, methanol, acetone, and ethyl acetate) of Roselle calyces had antimicrobial effect on multidrug-resistant Salmonella strains (Go´mez-Aldapa et al., 2017). Literature shows that calyces of Roselle contain different active compounds with antimicrobial action such as flavonoids (e.g., gossypetin) (Gomez-Aldapa et al., 2018) besides other known organic acids and phenolic compounds.

4.2.3 Influence of plant growth, age, and harvesting on Hibiscus sabdariffa quality The highest dry mass accumulation in the seed indicates the physiological maturity (PM) of Roselle (Copeland & McDonald, 1995). To properly preserve seeds for better germination, the right stage of PM needs to be observed. Previous data show that the maximum size and weight of the fruits and other floral parts are preceded by attainment of PM in Roselle (Rahman, Fakir, & Rahman, 2010). Moreover, a higher calyx yield in Roselle can be attained when the proper harvest time is observed. Hence the optimal time for the attainment of higher calyx yield is a crucial quality determinant. Furthermore, good nutrient and seed maturity depend upon the growth stages of the fruit after pollination and fertilization. Ali, Zain, and Latip (2019) argued that the wet red fleshy Roselle calyces must be

4.2 Quality and quality maintenance of the Hibiscus sabdariffa

harvested when the petals drop, but before the seedpods are thoroughly dried and dehisced (Monteiro et al., 2019). Therefore it is necessary that the capsule takes a shorter time after the seed ripens since the long capsules remain on the plants after seeds have begun to ripen, making the calyx more susceptible to sores, sun cracking, and affects quality. Therefore it would be prudent to harvest fleshy calyx and dry seeds for maximum yield simultaneously. The importance of harvesting before capsules dry and open thus making handling difficult leads to nonuniform drying (McClaleb, 1998) and this eventually leads to lowering quality of calyces. The period between flowering and complete maturity of calyces is estimated between 30 and 40 days, depending on variety and season (Castro et al., 2004). According to these authors, the determination of the calyx harvesting stage is crucial for the optimal production of Roselle calyx yield of higher and superior quality attributes (Castro et al., 2004). For a long time, Roselle growers have naturally harvested red fleshy calyces before the seeds dry and shatter. Notwithstanding the harvesting time, research has also shown that quality attributes such as distribution and amount of phenolic phytochemicals could be affected by environmental factors, type of cultivars, seasons, and post handling practices (Kim, Lee, Lee, & Lee, 2002; Ozlem, 2011). There are compelling references that point to the need to determine the effect of maturity stage on phenolic content in fruits and vegetables (Ogah, Watkins, Ubi, & Oraguzie, 2014). In sweet cherries, anthocyanins accumulation analyzed showed that, the total level of anthocyanins is relatively higher in ripe sweet cherries than the partially ripened ones (Goncalves et al., 2007). According to Mileti´c, Popovi´c, Mitrovi´c, and Kandi´c (2012), anthocyanins content in Prunus domestica L. (Plum) was significantly affected by maturity significantly affected maturity, suggesting that the anthocyanins accumulation occurred during fruit development. Obviously, the effect of maturity on quality is closely and positively related to the anthocyanins’ content in some fruits and negative in some cases. Information on the quality of Roselle in relation to the stage of maturity indicated that ascorbic and tartaric acids increased after the onset of flowering (Sulieman, 2014). They also revealed that the Roselle calyces’ acids and pigments increased in an amount up to the end of the fourth week, starting from the date of flower opening (Sulieman, 2014). Ibrahim, Karamalla, and Khattab (1971) further reported that Roselle’s total acidity started to decrease continuously after the fourth week from flowering, while the ascorbic acid of this fruit increased continuously until just before ripening. Moreover, significant differences were recorded in pH values among different plant and fruit growth stages and or calyces harvest (Mukherjee & Dutta, 1967).

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4.2.4 Antinutritional components and the prevailing factors on Hibiscus sabdarrifa quality Hibiscus is a plant with so many good and beneficial attributes, as already highlighted. However, some compounds significantly modified by the external environment can be detrimental to consumers. They include phytates, hydrogen cyanide(s), oxalates, nitrates, saponins, and tannins constitute the antinutrients. The work of Musa (2012) showed considerable amounts of cyanide in H. sabdariffa that varied with the position of the leaf. He reported higher cyanide concentrations in the basal than upper leaves (Musa, 2012). This could be due to the enzymatic creation of cyanide in fully developed leaves where metabolic activities are at maximum than the immature leaves (Musa, 2012). Anhwange, Ajibola, and Okibe (2006) also showed H. sabdariffa to have a hydrogen cyanide content of 0.29 mg/100 in the seed. However, this level was quite low, implying that the seeds were safe and suitable for the diet. Nitrate contents in different parts of H. sabdariffa are also a concern. Research shows increased nitrate content in basal leaves followed by middle and least in the upper leaves (Musa, Oladiran, Ezenwa, Akanya, & Ogbadoyi, 2013) which is an indication that increases in nitrate content were dependent on leaf age (Singh et al., 2015). The higher nitrate content in older leaves than younger ones (Musa et al., 2013) may be due to lower activity of nitrate reductase enzyme in the former than in the later (Musa et al., 2013). A lower enzyme activity in basal leaves reduces the rate of protein formation from nitrogen and may favor nitrogen accumulation and subsequent nitrate oxidation in leaf regions affected (Musa et al., 2013). Excess amounts of nitrate in leafy vegetables cause methemoglobinemia, gastric cancer, and many other diseases (Rao & Puttanna, 2000). Singh et al. (2015) observed the highest nitrate content in the calyx (89.34 mg/100 g) while the lowest in leaves (64.28 mg/100 g). However, new findings suggest nitrate as nitrous oxide source, an important molecule for the functioning of muscle cells (Lundberg, Carlstrom, Larsen, & Weitzberg, 2011). Another antinutrients associated with different plants like legumes are oxalate, and this has also been a concern in Roselle quality. Results from the estimation of antinutritional factors in parts of H. sabdariffa were shown by Singh et al. (2015), where maximum oxalate content in the calyx (4.39 mg/100 g) was reported, while leaf portion had the lowest level (3.16 mg/100 g). The problem is that oxalates bind calcium to form calcium oxalate that precipitates around soft tissues like kidney and causes kidney stones (Igile, Iwara, Mgbeje, Uboh, & Ebong, 2013). However, the oxalate content in the H. sabdariffa parts was below the established toxic level (Igile et al., 2013). The results also show phytate contents ranging from 1173.81 mg/100 g in immature seeds to 175.66 mg/l00 g in flower (Singh et al., 2015). Phytic acid in H. sabdariffa parts agrees with earlier reports on tropical leafy vegetables (Adeboye & Babajide, 2007). This is relatively high compared to the results by Anhwange et al. (2006), who reported 5.9 mg/100 g. Abu-Tarboush and Basher Ahmed (1996) revealed high phytate

4.3 Maintenance of quality of secondary metabolites

contents in H. sabdariffa compared to soybean, but after the defatting process, the levels were substantially in nontoxic range. It is paramount to note that high levels of phytic acid are of nutritional significance as it reduces the bioavailability of minerals (Hosain and Becker, 2001). However, the good thing is that phytates could be substantially reduced or even eliminated through specific processing techniques, some as simple as cooking and soaking (Yasmin, Zeb, Khalil, Paracha, & Khattak, 2008). The fruit also contains a high amount of saponin (145.0 mg/100 g) and it is low in mature seeds (14.74 mg/100 g). Anhwange et al. (2006) reported 2.2% saponin in the seeds of H. sabdariffa, which is in the range of those reported by Rodr´ıguez-Salinas et al. (2020). Though it favors plant defense mechanism, high saponin in the human diet may cause gastroenteritis associated diarrhea and dysentery. Saponins have a bitter taste and foaming properties and can trigger injuries to the digestive mucosa and hemolytic alterations in the blood (Mercedes, Carmen, Gemma, Mercedes, & Carmen, 1999). Also related to saponins are tannins. Tannin contents of H. sabdariffa seed were reported at 0.16% by Anhwange et al. (2006). The nutritional effect of tannins is mainly related to their interaction with protein (Hosain & Becker, 2001). The tannin content of the seeds was found to be in trace amounts by these authors. Generally, soaking and cooking processes have been reported to reduce the levels of tannins significantly.

4.3 Maintenance of quality of secondary metabolites of Hibiscus sabdariffa Roselle calyx is the part of the hibiscus plant that is of great interest. It is used in making fruit preservatives, jellies, and jam rich in pectin, anthocyanin color, and ascorbic acids. There are strong indications from previous work that Roselle calyces are used as quality sources of raw food colorants due to their high natural pigments traditionally or in industries. Also, these calyces’ anthocyanins possess antioxidative, antitumor, and anticarcinogenic activity (Tsai & Huang, 2004). The juices are mostly and consistently known for their color instability during storage, and this has been proven to correlate well with variations in anthocyanin contents (Vankar & Shukla, 2011). Unfortunately, there is a paucity of information on processing conditions to achieve the final stable product with better storage and longer shelf life (Gonzalez-Palomares, Estarro´n-Espinosa, Go´mez-Leyva, & Andrade-Gonza´lez, 2009). For instance, spray-drying to obtain Roselle powder is an alternative approach for using calyces. The dehydrated products are added to other foods to enhance storage, transportation, and shelf life (Gonzalez-Palomares et al., 2009). There is a need for standardization of such processes. Evidence implicates that the number of solids in suspension and dissolved oxygen are some of the major causes of anthocyanin instability (Cisse et al., 2012; Ndong, Faye, Bassama, & Cisse´, 2018). Degradation of anthocyanins in

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storage is significantly affected by extraction methods and temperatures during storage (Cisse et al., 2012; Ndong et al., 2018). There is some documentation on how to stabilize the anthocyanins to extend the juice’s shelf life. This is achieved through the concentration process by water evaporation that creates stable hibiscus extracts (Ndong et al., 2018). Pasteurization and food additives are preservation techniques for stabilizing food products in storage (Ndong et al., 2018). Furthermore, ascorbic acid and its salts have been used to sterilize and reduce yeasts, molds, and bacteria through the accumulation of microorganisms in the cytoplasm, leading to acidification of the cytosol (Ndong et al., 2018; van Beilen et al., 2016). Consequently, these help to preserve the organoleptic, nutritional, and various microbiological properties of the stored foods (Ndong et al., 2018). The dynamics of different components of the plant, particularly during processing, need in-depth understanding and attention. Apart from pasteurization and food additives, thermal processing and pH adjustments are usually applied unit operations in the food industry (He, Yuan, Zeng, Tao, & Chen, 2015). Incidentally, in industrial beverage preservation, the use of thermal processing is prevalent (Andres, Villanueva, Mateos-Aparicio, & Tenorio, 2014). However, thermal treatments contribute to organoleptic and loss of nutritional values and cause changes in phenolic compounds, ascorbic acids, and carotenoid, thereby contributing to reduced antioxidant capacity and other bioactivities (Wu, Yang, & Chiang, 2018). Anthocyanins are greatly volatile and susceptible to degradation by external factors like temperature, pH, oxygen, enzymes, and metals ion and other factors (Wu et al., 2018). Aside from affecting foods directly, anthocyanin degradation can produce aldehyde substances with benzene rings (Wu et al., 2018) that have detrimental effects on human health (Preedy, Watson, & Patel, 2013). There is no need to stress that thermal treatment can cause organoleptic and nutritional loss to the products (Wu et al., 2018), leading to reduced antioxidant capacity and other effects on their bioactivities (Andres et al., 2014; Wu et al., 2018). Moreover, color failing and off-flavor formation limit commercial products’ shelf life (Wu et al., 2018). Anthocyanin is an essential quality indicator of Hibiscus but is known to deteriorate, mostly manifested in different color hues rapidly. Luckily there are new technologies such as encapsulation that can be employed to extend many compounds’ shelf-lives including anthocyanin. Encapsulation is a technology aimed toward restriction and absorption of a biologically active compound on or inside solid particles (microspheres) or liquid vesicles to alleviate, organize, and guard the active compound and allow control of its release (Cavalcanti, Santos, & Meireles, 2011). This can be attained by enabling light and heat-labile molecules like many pigments, such as anthocyanins, to maintain their stability and enhance their shelf-lives and effects (Cavalcanti et al., 2011). Another way of improving the quality of Roselle is by drying, and this should be in a specialized way. It has been proven that drying Roselle extracts can produce a stable powder that has a long storage life. Of essence, the drying process should be infused with carrier agents that can entrap active agents within a carrier

4.4 Methods of measurements of bioactive compounds

material that enhances the delivery of the biomolecules and living cells into food products, as reported by Nedovic et al. (2011). The authors cautioned that the materials used for carrier agents should be food grade, recyclable, and capable to form a wall or barrier between the internal and surrounding areas (Ram´ırez et al., 2015). They proposed that the most common carrier agents to include are maltodextrin, gum arabica, and modified starch (Ram´ırez et al., 2015). The difference between a variable noncarrier agent and using maltodextrin as an agent can be viewed in the total color difference values. Complete color change in drying without carrier agent increased significantly, as demonstrated by Djaeni, Utari, and Kumoro (2017). The study confirmed that the drying temperature significantly affected the products’ color, mainly when applied in highly susceptible environmental conditions and/or processing carriers. At times, carrier agent’s gives appropriate and valuable protection of functional compounds used to control flavor, color, texture, and other preservation properties (Nedovic et al., 2011). Unfortunately, beyond a certain point, the higher the drying temperature, the more inferior the quality of Roselle as manifested by color hues.

4.4 Methods of measurements of bioactive compounds in Hibiscus sabdariffa A number of documented methods can be used for measurements of ingredients for quality standard references. For instance, Singh, Singh, Ashish, and Salim (2013) made an attempt to determine the quality of Roselle using different measurement options. They reported on total phenols, antioxidants, total carotenoids, and ascorbic acids as some key determinant bioactive components considered as quality indices of Roselle. The high antioxidant activity of the leaves of H. sabdariffa was estimated with 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical method (Singh et al., 2015; Wang et al., 2014). This method is an easy, rapid, and sensitive way to evaluate the free radical scavenging capacity of plant extracts (Lim, 2016). It accepts an electron or hydrogen radical from an antioxidant compound ¨ ksal, GU ¨ LC¸˙IN, Beyza, Sarikaya, & to become a stable diamagnetic molecule (KO • Bursal, 2009). The reduced DPPH radical was observed by the decrease in absorbance at 517 nm (Jiang, Zhan, Liu, & Jiang, 2008), which disappears with an increase in enzyme concentration in solution through addition of antioxidant-rich plant extracts (Vankar and Shukla, 2011). Apart from DPPH method, other spectrophotometric assays can be used to determine the bioactive compounds in H. sabdariffa (Wang et al., 2016). To appraise the similarity of the two most common radical scavenging assays using 2,20 -azino-bis-3-ethylbenzthiazoline-6-sulfonic acid (ABTS) and DPPH radical, the 50 most popular antioxidant-rich fruits, vegetables, and beverages in the US diet were identified and analyzed for their antioxidant capacities, total phenolics, and flavonoids content (Floegel, Kim, Chung, Koo, & Chun, 2011). Other

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assays available use ferric reducing ability of plasma (FRAP; Floegel et al., 2011; Villani, Juliani, Simon, & Wu, 2013) and oxygen radical absorbance capacity (ORAC; Floegel et al., 2011; Shukla, 2020; Wang & Zhu, 2017). These assays use a similar principle for generating colored radical or redoxactive compound and have the ability to scavenge the radical or reduction of the redox-active compound that is monitored by spectrophotometer (Floegel et al., 2011) through the application of the right standard that quantifies the antioxidant capacity (Gulcin, 2020). Some examples of standards used in the quantification of antioxidants are vitamin C equivalent antioxidant capacity (VCEAC) and trolox equivalent antioxidant capacity (Chen & Yen, 2007). The two assay methods that are employed to quantify antioxidants are (1) electron-based transfer that entails a reduction of colored oxidants, for example, in ABTS, DPPH, and FRAPs and (2) hydrogen atom transfer like ORAC assay, where the antioxidants strive with the substrates for thermally generated peroxyl radical (Floegel et al., 2011). The ABTS assay operates on the generation of a blue/green ABTS 1 that undergoes a reduction process by antioxidants (Adelakun, Kudanga, Green, Roes-Hill, & Burton, 2012), whereas the DPPH assay thrives on reducing the purple DPPH to 1,1-diphenyl-2-picryl hydrazine (Cheng, Lai, Lin, & Sakata, 2016). The two assays are useful and thus the most popular; nevertheless, there are limitations on their use since they are nonphysiological radicals (Floegel et al., 2011). Comparatively, ORAC assay can detect chemical changes in fluorescent molecules that are caused by an attack of free radicals. The assay is based on peroxyl radical with a reflection of physiological relevant perturbations (Floegel et al., 2011). Conversely, the FRAP assay differs from others as it does not involve free radicals but the reduction process of ferric iron (Fe31) to ferrous iron (Fe21) (Floegel et al., 2011). As much as there are some spectrophotometric assays in existence, most of the analyses with H. sabdariffa have been accomplished by DPPH assay. Antioxidant activity changes with the concentration of extracts (Singh et al., 2011), as observed in the study by Ali et al. (2019). H. sabdariffa parts also showed an increasing trend for antiradical capacity with an increase in the concentration of the extracts. Investigation of antioxidants through the approach of DPPH scavenging assay or inhibition percentage and comparing it with ascorbic acid as a standard of reference had been previously undertaken by Ali et al. (2019). They looked at the antioxidant activities from varied maturation stages of Roselle. Chen, Lin, Chen, and Chiang (2019) showed that the sequence for DPPH free radical scavenging activity of the plant extract improved with the maturity period thus decreasing from old to the young (Lim, 2016). The DPPH free radical scavenging activity of the Roselle calyx revealed a gradual ¨ ksal et al., 2009). Ali et al. (2019) increase as the calyx matured (KO reported that a corresponding increase in total phenolic contents correlated with antioxidant activities throughout the ripening period. According to Siti Aishah, Rohana, Masni, and Jalifah (2019), phenolic content measurements

References

like anthocyanins serve as indicators to establish the quality and processing of Roselle products. Their report showed profiles of delphinidin-3sambubioside and cyanidin-3-sambubioside (Lim, 2016). The two anthocyanins, delphinidin-3-sambubioside and cyanidin-3-sambubioside (Lim, 2016; Ojeda et al., 2010), were high during all the development stages of the calyx (Su et al., 2018). The presence of these anthocyanins in Roselle has been reported by others (Bernal, Orduz-Diaz, & Coy-Barrera, 2016; Jafarian, Mortazavi, Kenari, & Elhami Rad, 2014; Kouakou et al., 2015). Though considered less superior to HPLC, the DPPH method is technically simple (Locatelli et al., 2009). It requires only a UV
vis spectrophotometer to perform: in the presence of a hydrogen/electron donor (free radical scavenging antioxidant), the absorption intensity is reduced, and the radical solution is discolored according to the number of electrons captured (Locatelli et al., 2009). Nonetheless, electron paramagnetic resonance spectroscopy is preferred to assess the DPPH radical in the case of antioxidant compound such as carotenoids with spectra that overlaps DPPH at maximum absorbance (Kelebek & Selli, 2014; Locatelli et al., 2009). Gonza´lez-Mun˜oz, Quesille-Villalobos, Fuentealba, Shetty, and Ranilla (2013) found that ABTS assay is more useful than DPPH assay for detecting antioxidant capacities in a variety of foods. Xia, Yuxiao Xing, and Kan (2019) compared ABTS and DPPH for analyzing the antioxidant capacities, total phenolics, and flavonoids content in 50 most popular antioxidant-rich fruits, vegetables, and beverages in the US diet (Floegel et al., 2011). Antioxidant capacity showed a strong positive relationship when both assays were compared (Floegel et al., 2011). Moreover, antioxidant capacity detected by ABTS assay was stronger and positively associated with the ORAC from USDA database, phenolics, and flavonoids content (Floegel et al., 2011).

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Composition of Hibiscus sabdariffa calyx, pigments, vitamins

5

Abdalbasit Adam Mariod1,2, Haroon Elrasheid Tahir3 and Gustav Komla Mahunu4 1

Indigenous Knowledge and Heritage Center, Ghibaish College of Science & Technology, Ghibaish, Sudan 2 College of Sciences and Arts-Alkamil, University of Jeddah, Alkamil, Saudi Arabia 3 School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China 4 Department of Food Science and Technology, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana

Chapter Outline 5.1 Introduction ................................................................................................... 69 5.1.1 Plant description ...........................................................................69 5.1.2 The chemical composition of Hibiscus sabdariffa calyx .....................70 5.1.3 Composition of Hibiscus pigments ..................................................71 5.1.4 Composition of Hibiscus sabdariffa vitamins ....................................72 5.1.5 Parts used medically .....................................................................72 5.1.6 Pharmacological and therapeutic properties of hibiscus ....................73 References ............................................................................................................ 74

5.1 Introduction 5.1.1 Plant description The Hibiscus sabdariffa (HS) plant is a small shrub, reaching a height of about 2 m. The stem and branches of the plant are green in color, impregnated with redness, and the leaves are simple with long necks and serrated edges, reddish-green in color, and resemble the palm (El Shazly et al., 2018). The plant bears plump, fleshy, with beautiful-looking flowers that emerge from the tip of the leaf stalk; flower parts are thick and imbued with a dark red color. As for the fruits in the form of capsules, inside them are a number of brown seeds, which are spherical in shape and wrinkled on the surface. The plant is originally from India, but it is cultivated in many countries, especially Sudan. It was known in the past among the Pharaohs, Arabs, and Indians, and it had multiple uses and recipes, some of which are still used today Roselle (Hibiscus sabdariffa). DOI: https://doi.org/10.1016/B978-0-12-822100-6.00002-1 © 2021 Elsevier Inc. All rights reserved.

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FIGURE 5.1 Hibiscus sabdariffa leaves and calyx.

(Mohamed, Sulaiman, & Dahab, 2012). H. sabdariffa can usually be extracted with hot or cold water to obtain a red sour drink, can be added to soups, or it can be cooked on its own with butter or oil. The hibiscus flowers are washed and placed in warm water and heated over a fire without boiling and left covered for half an hour, then filtered and served either cold or hot, and as desired, honey or sugar can be added to it with or without lemon (Salman et al., 2017). Seeds, leaves, fruits, and roots of Roselle are utilized in different foods just as in natural medication as a potential nonpharmacological treatment. Various concentrates from Roselle assume a pivotal function in treating distinctive clinical issues including numerous cardiovascular problems, helminthic sickness, and malignant growth. The plant additionally goes about as an antioxidant and is utilized to control body weight (Singh, Khan, & Hailemariam, 2017) (Fig. 5.1).

5.1.2 The chemical composition of Hibiscus sabdariffa calyx The hibiscus calyx contains glycosides in addition to colorants, calcium oxalate salts, and vitamin C. The hibiscus is colored dark red in the acid medium due to the presence of betacyanin compounds and it contains gels (Ademiluyi, Oboh,

5.1 Introduction

FIGURE 5.2 Dried Hibiscus sabdariffa calyx.

Agbebi, & Akinyemi, 2013). The plant sepals contain organic acids such as malic, tartaric, citric, glycoside, and hepiscin chloride, some mucous materials, a stable oil in the seeds, as well as glycosides, salts (calcium oxalate, potassium salts (K), magnesium salts, iron salts), and vitamin C (Wong, Yusof, Ghazali, & Che Man, 2002). Hibiscus flowers are acidic in taste because they contain a lot of natural acids, and they are rich in iron, vitamin (B) complex, calcium, phosphorous, and the leaves contain small amounts of cellulose (Mahadevan & Kamboj, 2009). Storing hibiscus after harvest is a challenge because it is perishable, and there are different methods of drying, but it is difficult to have an alternative for postharvest use in foods without compromising the quality and effectiveness of antioxidants. The most important of these methods are sun drying, infrared drying, and freeze-drying (Kumar, Prabhakaran, Shetty, & Giridhar, 2014). Flavonoids, anthocyanidins, triterpenoids, steroids, and alkaloids were extracted and determined from Roselle (Manita-Mishr, 1999) (Fig. 5.2).

5.1.3 Composition of Hibiscus pigments Food colorants are either natural or artificial. Natural colorants are extracted from renewable sources such as plants, insects, algae, etc., while artificial colorants are manufactured or chemically synthesized in the form of dyes and are most commonly used in the food, pharmaceutical, and cosmetic industries. As a result of some restrictions and the global tendency to consume natural products, the demand for natural colorants has increased significantly (Huck and Wilkes, 1996). The anthocyanin pigments were extracted from the hibiscus and the basic composition of these pigments was identified by high performance liquid chromatography (HPLC). These dyes were used as natural alternative colorants in some processed foods such as hard candy (Khoo, Azlan, Tang, & Lim, 2017). Anthocyanin pigment, which is a compound or colored pigment, may be red, purple, or blue. . . etc., and is found in flowers, fruits, stems, roots, and leaves,

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and it is mentioned that most of the colors of flowers and fruits are due to the anthocyanin pigment, although tomatoes and some yellow flowers are colored due to carotenoid pigments (Khoo et al., 2017). It is noteworthy that the anthocyanin pigment is usually associated with sugar molecules with glycoside bonds, and when removing the sugars, the remaining part is called anthocyanidins, and the most famous of its compounds are cyanidin, delphinidin, peonidin, etc. Thus the color of anthocyanin depends on the substitution groups present in the B-ring in the composition of this anthocyanin dye. Anthocyanin helps in attracting insects and birds to facilitate the pollination and fertilization process, plays an important role in plant disease resistance (Miller, Owens, & Rørslett, 2011). Selim, Khalil, Abdel-Bary, and Abdel-Azeim (2008) produced a dry red powder from the Roselle calyces of the hibiscus plant and used it to color some food products using the microencapsulation technique. The research team measured the stability of the encapsulated dyes during storage under different aqueous activities. The results obtained confirmed that most of the encapsulated dyes showed stability during the storage period and retained their anthocyanin content. The research results also showed that adding hibiscus dyes coated to berries, jam, and hard candy resulted in an acceptable product similar to the controls. A study was conducted to evaluate the stability of hibiscus extract at different light and temperatures. Then the anthocyanin pigment, phenolic compounds, and color were measured. A gelatin product was developed with the addition of hibiscus extract and a sensory analysis test was performed, the study showed the effect of light and temperature factors on anthocyanins, phenolic compounds during storage. The highest stability of the anthocyanin pigment and the phenolic compounds, respectively, was observed at a temperature of 4 C and in the absence of light, a loss of stability was observed at 25 C with light. Regarding the pH, it was observed that the change in pH affected the absorbance and color in the hibiscus extract. Finally, when adding hibiscus extract with a concentration of gelatin, it found favor with me and exhibited seven shades of color and general acceptance (Para´ıso et al., 2020).

5.1.4 Composition of Hibiscus sabdariffa vitamins The H. sabdariffa is rich in carotene, riboflavin, anthocyanins, ascorbic acid, niacin, and vitamin C. The young leaves and tender stems of Roselle are consumed raw as a green vegetable (Singh et al., 2017). H. sabdariffa blossom contains ascorbic acid that is 260
280 mg in 100 g. Roselle can be handled into sirup, concentrates of Roselle should be possible by warming or without warming (Mukaromah, Susetyorini, & Aminah, 2010). The sour taste of Roselle is related to ascorbic acid content and boiling of Roselle for 10 minutes reported to increase the vitamin C but more boiling time will decrease vitamin C.

5.1.5 Parts used medically The used parts of the hibiscus plant are the flowers and leaves, and they are collected in every season, and the best part of the flower is the sepals that surround

5.1 Introduction

FIGURE 5.3 A refreshing Roselle drink.

the flower and these are collected when they start drying to either dark red or light red color (Mahbubul Islam, 2019). The Roselle drink is very famous in many countries and it is a food trend in muslim countries during fasting as it is well known to quench thirst (Fig. 5.3).

5.1.6 Pharmacological and therapeutic properties of hibiscus Some scientists have reached the extreme efficacy of the hibiscus plant as an antioxidant that lowers high blood pressure, increases the speed of blood circulation, and strengthens the heartbeat. HS can be used as a moisturizer and tonic for the body to prevent the feeling of thirst, it is useful on hot days and during fasting. Because it contains a high content of vitamin C, it can help in treating cold, cough, and high fever. In relation to its antioxidant property, it is recommended while taking chemotherapy for cancer patients, as well as used to fight cancer as a disease (Da-Costa-Rocha, Bonnlaender, Sievers, Pischel, & Heinrich, 2014). HS contains disinfectants and kills microbes, which makes it useful in treating fevers and microbial infections and for getting rid of tapeworms and roundworms (Ascaris) in the intestine. HS seems to be useful for contractions of the uterus, stomach, and intestines and eliminates pain during the menstrual cycle. Therefore

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girls who suffer from period pain are advised to take a cup twice a day, days before menstruation. HS is used as a natural dye for medicines, food, and cosmetics, such as increasing the fixation of hair color and giving it a bright color (Jabeur et al., 2017).

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53.

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Para´ıso, C. M., dos Santos, S. S., Ogawa, C. Y. L., Sato, F., dos Santos, O. A. A., & Madrona, G. S. (2020). Hibiscus sabdariffa L. extract: Characterization (FTIR-ATR), storage stability and food application. Emirates Journal of Food and Agriculture, 32 (1), 55
61. Available from https://doi.org/10.9755/ejfa.2020.v32.i1.2059. Salman, A., Abdullah, I., Saib, I., Abdulaziz, B., Abdullah, K., Yahya, K., . . . Saleh, W. (2017). Acute effect of drinking cold hibiscus beverage on blood pressure in adult females: A randomized controlled trial. International Journal of Academic Scientific Research, 5(1), 26
34. Selim, K. A., Khalil, K. E., Abdel-Bary, M. S., & Abdel-Azeim, N. A. (2008). Extraction, encapsulation and utilization of red pigments from Roselle (Hibiscus sabdariffa L.) as natural food colorants. In Alex. J. Fd. Sci. & Technol. Special Volume Conference, pp. 7
20, Mar. 2008. Singh, P., Khan, M., & Hailemariam, H. (2017). Nutritional and health importance of Hibiscus sabdariffa: a review and indication for research needs. J Nutr Health Food Eng, 6(5), 125
128. Available from https://doi.org/10.15406/jnhfe.2017.06.00212. Wong, P.-K., Yusof, S., Ghazali, H. M., & Che Man, Y. B. (2002). Physico-chemical characteristics of Roselle (Hibiscus sabdariffa L.). Nutrition & Food Science, 32(2), 68
73. Available from https://doi.org/10.1108/00346650210416994.

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Hibiscus sabdariffa: protein products, processing, and utilization

6

Mildred Osei-Kwarteng1, Joseph Patrick Gweyi-Onyango2, Gustav Komla Mahunu3, Haroon Elrasheid Tahir4 and Maurice Tibiru Apaliya5 1

Department of Horticulture, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana 2 Department of Agricultural Science and Technology, Kenyatta University, Nairobi, Kenya 3 Department of Food Science and Technology, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana 4 School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China 5 Department of Hotel Catering and Institutional Management, Cape Coast Technical University, Cape Coast, Ghana

Chapter Outline 6.1 Introduction ................................................................................................... 77 6.2 Protein products ............................................................................................. 79 6.2.1 Seeds ...........................................................................................79 6.2.2 Leaves .........................................................................................79 6.2.3 Calyces and general flowers ...........................................................80 6.3 Processing of protein products ........................................................................ 80 6.3.1 Types of processing methods ..........................................................81 6.3.2 Quality of proteins from different processing methods ......................82 6.4 Utilization of Roselle protein products ............................................................. 84 6.5 Conclusion ..................................................................................................... 85 References ............................................................................................................ 85

6.1 Introduction The various parts of the Roselle plant contain rich proteins. The protein content in the seeds is between 13% and 35.4%, which is higher than that of the other parts such as the leaves (3.5 g/100 g) and the calyces (2 g/100 g) (Naturland e.V., 2000). Ansari, Eslaminejad, Sarhadynejad, and Eslaminejad (2013) also reported

Roselle (Hibiscus sabdariffa). DOI: https://doi.org/10.1016/B978-0-12-822100-6.00012-4 © 2021 Elsevier Inc. All rights reserved.

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4.71 g/100 g, 6.45 g/100 g, and 9.87 g/100 g of dry matter protein content for the red and green calyces and flowers, respectively. There are papers that show that change of diet from mostly animal-based proteins toward plant-based protein products is crucial for food security and sustainability (de Boer & Aiking 2017, Aiking & de Boer 2020). Plant proteins are easy to assess as compared to animal proteins in relation to the food chain and web, and it is not surprising that plants (including Roselle) are therefore the major sources of low-cost protein (Halimatul, Amin, Mohd-Esa, Nawalyah, & Muskinah, 2007; Tounkara, Amza, Lagnika, Le, & Shi, 2013a; Tounkara, Sodio, Amza, Le, & Shi, 2013b). Essentially, plant-based proteins have been used to replace the functional and nutritional properties of skeletal muscle proteins in a variety of processed meat products, and this is related to the growing realization of nutraceutical benefits derived from plants as opposed to animalbased protein sources (Egbert & Payne, 2009). Notwithstanding, plant-based proteins tend to have lower anabolic effects than animal proteins because their digestibility and essential amino acid (e.g., leucine) content are low and are deficient in essential amino acids like lysine and sulfur amino acids (Berrazaga, Micard, Gueugneau, & Walrand, 2019). However, the lower anabolic capacity of plant-based proteins can be improved by fortifying them with specific essential amino acids, selective breeding, blending several plant protein sources, and blending plant with animal-based protein sources (Berrazaga et al., 2019). The increase in consumers’ demand as a result of health and environmental concerns and vegetarianism trends has led to a renewed interest in plant protein. A widely known source of plant proteins are derived from Soybean (Glycine max L.) (Messina, 1999) and wheat, although there is a growing trend that shows that other future sources of commercially available plant proteins may include pea, potato, corn, canola, rice, and other proteins from legumes and oilseeds (Egbert & Payne, 2009; Petrusa´n, Jawel, & Huschek, 2016). Apparently, there is a worldwide need for much cheaper protein sources. Hence, efforts have been made to develop new sources of easily accessible, digestible (Salah & Hayat, 2009) and low-cost protein foods from environmentally wide adaptable plants such as Roselle which are abundant especially in Africa (Halimatul et al., 2007). This is because protein fractions, isolates, or concentrates obtained from this plant are promising alternative sources of lowcost protein in dietary supplements or ingredients for the food industry. Henceforth its adoption will alleviate the problem of protein scarcity especially in underdeveloped or third world countries (Halimatul et al., 2007; Tounkara et al., 2013a,b). This chapter aimed at filling the knowledge gap through the review of information on the protein products of the Roselle plant, the processing methods used and influence of processing methods on the said products, and how the protein products are utilized.

6.2 Protein products

6.2 Protein products Protein can be obtained from the various parts (seeds, kernels, calyx, and leaves) (Ayo-Omogie & Osanbikan, 2019; Jeannett et al., 2020; Mabrouk et al., 2018) of the Roselle plant but the seeds are the main source of proteins (Mokhtari, Zarringhalami, & Ganjloo, 2018). Key protein fractions, protein isolates, or concentrates emanating from Roselle seeds (RS), and other parts may constitute a potential and most viable alternative sources of low-cost protein substitute in dietary supplements or ingredients for food industry (Rimamcwe, Chavan, Dalvis, & Gaiwad, 2017).

6.2.1 Seeds Earlier studies have revealed that RS could be used as possible sources of proteins (Al-Wandawi, Al-Shaikhly, & Abdul-Rahman, 1984; El-Adawy & Khalil, 1994; Hainida, Ismail, Normah, & Norhaizan, 2008; Halimatul et al., 2007; Shaheen, El-Nakhlawy, & Al-Shareef, 2012). They are nutritionally rich in proteins (13%
35.4%) (Anel, Thokchom, Subapriya, Thokchom, & Singh, 2016; AyoOmogie & Osanbikan, 2019; Juhari & Petersen, 2018) and essential amino acids (39.47%) (Mabrouk et al., 2018; Amin, Alkaabi, Al-Falasi, & Daoud, 2005; Tounkara, Bashari, Le, & Shi, 2014; Halimatul et al., 2007), with methionine and cystine being the limiting amino acids (Ayo-Omogie & Osanbikan, 2019; Mabrouk et al., 2018). Comparatively, these seeds are among the highest proteincontaining seeds (Shaheen et al., 2012). Relative to seeds like pigeon peas, groundnut, cowpea, black seed, sunflower seed, melon seed, chickpea, and soybean, RS have higher protein content (32.28%
35.4%), whereas their essential amino acid profile compares to that of soybeans (Ayo-Omogie & Osanbikan, 2019). The crude protein (CP) content of the seeds ranges from 21.85% to 30.6% (Hainida et al., 2008, Nzikou et al. 2011). Other protein products of RS include seed cake, defatted seed meal, and protein concentrates from defatted seed. The aforementioned RS products are promising as value-added nutritional foods and livestock rations (Dhar et al., 2015). Notably, Roselle protein isolates have more protein content (88.15%) compared to other Roselle products such as defatted flour (50.63%) and protein concentrate (62.24%) (Abu-Tarboush, Ahmed, & Al Kahtani, 1997). Indigenous “Furundu” is also a meat substitute prepared by cooking RS and fermenting it for 9 days. Fermentation of the seeds increases the total protein content (Yagoub, Mohamed, Ahmed, & El Tinay, 2004).

6.2.2 Leaves Roselle leaves contain proteins aside from other nutrients (Anel et al., 2016; Mahadevan & Pradeep, 2009). The fresh leaves contain 3.3
3.5 g/100 g protein

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(Duke & Atchley, 1984; Ismail, Ikram, & Nazri, 2008; Singh, Khan, & Hailemariam, 2017) and between 1.7% and 3.2% in 100 g of Roselle (Jeannett et al., 2020). The specific amino acids found in the leaves include phenylalanine, glutamic acid, lysine leucine, and arginine (Hainida et al., 2008; Ismail et al., 2008). Additionally, Anel et al. (2016) reported that red calyx plant leaves had higher protein than those from the yellow calyx plant.

6.2.3 Calyces and general flowers Roselle is well known and valued for edible calyx that can be utilized in many ways, including beverages. In most varieties, the calyx is the red colored pointed pods that protect and support the plant (Islam, 2019). Depending on the environmental conditions and varieties in question, calyces of Roselle taste sweet and should be picked 10
15 days after they lose their blooms, or else it will taste more tart (Cobely, 1976). Nutritional properties of Roselle calyces previously reported by Duke (1983) and Mat Isa, Isa, and Abd Aziz (1985) showed that 100 g of fresh Roselle calyces contained 1.9 g protein. The analysis of the calyces showed that they contain proteins (1.15%
17.40%) and essential amino acids with the exception of tryptophan (Cid-Ortega and Guerrero-Beltra´n, 2015; Ismail et al., 2008; Mahadevan & Pradeep, 2009). The reported wide range of protein content could be explained by the type of variety, genetics, ecology, the environment of growth, and the harvesting condition applied (Atta et al., 2013; Ismail et al., 2008). For instance, research carried out by Atta et al. (2013) in Niger showed that protein content in calyces for ecotype (E9) (52 mg/g dry weight) was approximately halved compared to those of ecotype (E3) and (E7).

6.3 Processing of protein products The use of plant proteins is limited by the presence of antinutritional agents and deficiency in some essential amino acids. These factors (together with other nutritional, processing, and physicochemical aspects) are of importance when considering the utilization of plant protein in human consumption (Ahmed, 1998). The bitter taste of raw RS is commonly associated with antinutritional constituents which tend to reduce by activation through processing such as heating (either moist or dry) or soaking (Ayo-Omogie & Osanbikan, 2019; Yacoub & Abdalla, 2007). While RS have cardio-protective effect, raw seeds are toxic as a result of transaminases and creatinine. A study showed that the comparative weight gain in the kidneys and livers of rats was compromised when seeds were consumed raw (Ghislain, Etengeneng, Sonia, Noelle, & Gouado, 2014). It is therefore imperative that Roselle products should be subjected to various processing methods to reduce

6.3 Processing of protein products

the phytotoxic compounds so as to improve the palatability, general uptake and digestibility, and overall quality.

6.3.1 Types of processing methods The composition of the end product depends on the appropriate processing method (Choong, Yousof, Wasiman, Jamal, & Ismail, 2016). Some of the reported processing methods for proteins from especially RS and other parts include oven roasting, microwave roasting, roasting and grinding into powder, boiling (Amon, Soro, Assemand, Due´, & Kouame´, 2011; Mariod et al., 2013; Mariod, Mirghani, & Hussein, 2017), fermentation techniques (Lakra and Sehgal, 2011; Yagoub & Mohammed, 2008), cooking, sprouting or germination, soaking (Ismail et al., 2008; Yagoub & Mohammed, 2008), and dehulling (Ayo-Omogie & Osanbikan, 2019).

6.3.1.1 Processing of calyx proteins Calyx is the most used part of the Roselle plant. It may be dark red, red, or green (Schippers, 2000). Extraction of free amino acids can be done as recommended by Moore et al. (1958) with modifications. The protein (N 3 6.25) can also be determined using the methods described by the AOAC (1998). The protein content of different colored calyces either decreases or increases in response to processing methods. Adanlawo and Ajibade (2006) found that when different colored calyces (red or green) were soaked overnight in wood ash, the protein content of the red calyx increased from 4.71 to 6.64 g/100 g, whereas that of the green calyx reduced from 6.45 to 5.57 g/100 g. Roselle calyx can also be fermented either naturally, or using coconut husk ashes and cocoa pod ashes, or using Gmelina tree ashes for 72 hours. The protein content of Roselle calyx was enhanced when it was fermented and neutralized with different concentrations of sodium sesquicarbonate (“trona”; salt) (Ojokoh, 2010).

6.3.1.2 Processing of leaf proteins Shade or sun-drying methods are simple but can influence the quality composition of proteins in Roselle plant parts (Garti, Garti, & Nyarko, 2019). The concentrations of nutrients were generally higher in sun-dried leaves than in the shadedried leaves. This observation confirms the results of Chege, Kuria, Kimiywe, and Nyambaka (2014) who showed that solar drying results in a relatively stable concentration of nutrients such as iron, zinc, and beta-carotene. This may be due to the loss of water which increases nutrients density. Thus the concentration of nutrients in leave is influenced by drying time and temperature. A dehydration process that rapidly removes moisture from a plant tissue inactivates the plant enzymes and reduces the loss of nutrients. Drying followed by milling is another processing technique. The leaves are oven-dried overnight at 65 C and then finely milled by the use of a multibeads

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shocker (Yasui Kikai, model MB500E). This is followed by the determination of total Ca, K, Mg, P, and N using appropriate laboratory analysis procedures. Total N content is determined by colorimetric method and then converted to protein.

6.3.1.3 Processing of seed proteins The methods of processing RS proteins include boiling, drying, boiling and sundrying, raw freeze-drying (Hainida et al., 2008; Halimatul et al, 2007; Ismail et al., 2008), and fermentation (Yagoub & Mohammed, 2008). Protein contents were high when all these processing methods were used. However, the percentage of protein in the raw-freeze-dried processed seeds (35.4%) was higher and statistically different (p , 0.05) relative to sun-dried (33.5%) and boiled-sun-dried seeds (30.6) (Hainida et al., 2008). Similarly, RS can be processed into powder after drying and boiling (Ghislain et al., 2014; Ismail et al., 2008) or roasting and grinding (Ismail et al., 2008). Their work showed that raw RS had 22% of CP, which was increased in the range of 23% and 24% with roasting, whereas soaktoasting of seeds gave values ranging between 25% and 26% when the same seeds were boiled. Moreover, results from Bozinviya, Yahaya, and Nyameh (2010) revealed 22%, 24%, and 24% CP for raw, soaked, and toasted seeds, respectively. A general increase in CP after boiling, toasting, or soak-toasting as compared to raw products is not uncommon in RS. Akinmutimi (2001) found that CP levels of heat-treated legume seeds increased slightly. Ghislain et al. (2014) on the other hand observed that boiling and soak-toasting enhanced RS digestibility and quality. The authors also found that net protein ratio (NPR), food intake and utilization efficiency, protein efficiency ratio (PER), weight gain, and true digestibility (TD) values were significantly improved by boiling and soak-toasting when compared with raw seed diets of rats. Similar results had been previously reported in rats by Halimatul et al. (2007) and Duwa, Oyawoye, and Njidda (2012) in broilers.

6.3.2 Quality of proteins from different processing methods Studies have shown that processing methods can affect the quality of Roselle protein products (Hainida et al., 2008). Ismail et al. (2008) highlighted three basic parameters that determine the quality of a protein source. These are (1) bioavailability of amino acids, (2) amount of indispensable amino acids, and (3) digestibility of protein. Protein quality refers to the dietary value of the protein which depends on the amino acid content, bioavailability of amino acids, digestibility, absorption, and minimal obligatory oxidation rates of the proteins (Halimatul et al., 2007; Ismail et al., 2008; Tounkara et al., 2013a,b; Hoffman and Falvo, 2004). Poor quality proteins are lacking in one or more amino acids, whereas good quality proteins are easily digestible and contain the required quantities of essential amino acids for humans (Ismail et al., 2008).

6.3 Processing of protein products

Initially 18 amino acids were identified in mature RS with values ranging from 1.35 to 23.45 g/16 g of nitrogen (Al-Wandawi et al., 1984). The ratio of amino acids is used in differentiating similar proteins or to highlight the reduction in specific amino acids after chemical modification of the protein after processing (Ismail et al., 2008). The quality of plant proteins also depends on the source and dietary mixture, whereas their digestibility depends on the food preparation method (Young & Pellet, 1994). Also, the critical functional properties (i.e., foam stability, gelation, emulsification, and solubility properties) of the proteins are very vital in defining protein quality and subsequent use of the protein for the production of new food and nonfood products by industries (Salah & Hayat, 2009). Hainida et al. (2008) revealed that the extraction method to obtain Roselle proteins has no negative effect on the amino acid profile. Arginine (34 g/100 g), lysine (14 g/100 g), leucine (15 g/100 g), and glutamic acid (24 g/100 g) were found to be high in RS products. Several techniques are employed in the quality determination of a protein. These include PER, biological value (BV), protein digestibility corrected amino acid score, and net protein utilization (Hoffman and Falvo, 2004). Henceforth, a protein quality test estimates or measures the nutritional value (i.e., dietary essential amino acid content) of food proteins and their availability for cell growth and maintenance (Halimatul et al., 2007). For instance, in order to utilize seed proteins as a functional ingredient, some quality parameters such as the biological PER, NPR, and TD values should be determined (Halimatul et al., 2007). Moreover, the quality of a protein of a plant can be assessed by comparing the amino acid composition with that of the World Health Organization (WHO) standard protein (Ismail et al., 2008).

6.3.2.1 Seed proteins RS proteins are of good quality, especially the proteins of the boiled seeds (BS) (Ghislain et al., 2014). In protein value assessment it is inappropriate to use human subjects due to the set upregulatory procedures; hence, several in vitro and in vivo bioassays have been developed for use in protein quality evaluation (Tounkara et al., 2013a,b). In their study, Tounkara et al. (2013a,b) used the amino acid composition as the basis for estimating the nutritional quality of RS protein isolates. They showed that in all the samples, the ratio of essential to total amino acids (E/T) was above the level of 36% recommended by FAO/WHO/UNU, whereas the prolamin fraction had a high ratio of 41.22%. Generally, a PER below 1.5 implied a protein of low or poor quality, whereas PER between 1.5 and 2.0 indicated an intermediate protein quality, and a PER above 2.0 means protein of high-quality (Tounkara et al., 2013a,b). They also found that the PER values of protein isolates from Roselle seed (RSPI) and its fractions were quite satisfactory compared with a standard casein PER of 2.5. Halimatul et al. (2007) also investigated the bioavailability and digestibility of RS proteins as affected by two thermal treatments (drying and boiling) in Sprague

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Dawley rats for 4 weeks so as to recommend the seeds for human consumption or industrial uses. They found that rats fed with boiled seeds (BS) ate more and had a highly significant (p , 0.05) weight gain than those fed with dried seeds (DS). However, PER and NPR of BS did not differ significantly (p , 0.05) with that of the DS. Consequently, they concluded that the protein quality of DS (100 C at 30 minutes) was similar to that of BS because the BV, apparent digestibility (AD), and TD of the two diets did not significantly differ. Likewise, other investigators have also showed that the protein quality of dried RS was similar to the protein quality of boiled RS (Hainida et al., 2008; Ismail et al., 2008).

6.3.2.2 Leaf proteins The CP in Roselle leaves (27.0%
28.0%) was higher than that in the calyces (16.0%
17.5%, Muhammed Amin & Ali Kanimarani, 2020) other reports also indicated 3.3 g/100 g of protein in the leaves. Protein in the dried leaves of Roselle was 5.37% (Balarabe, 2019). The amount of CP (28.7%) in Roselle leaves is comparable to that of Moringa olerifera (Teixeira, Carvalho, Neves, Silva, & Arantes-Pereira, 2014). Vegetable proteins are inexpensive food and can provide the nutritional needs of the vulnerable, particularly in developing countries. CP is a nutrient essential for the growth and maintenance to the body.

6.3.2.3 Calyx proteins Calyx color influences how a processing method affects protein content of the calyx. When green and red colored calyces were soaked (steeped) in wood ash overnight and washed thoroughly prior to its use for soup preparation, it was found that the protein in the red calyx increased (4.71
6.64 g/100 g), whereas that of the green calyx decreased (6.45
5.57 g/100 g) after the overnight steeping (Adanlawo & Ajibade, 2006). There are observable changes that have demonstrated the effects of cultivar in terms of the proximal composition of Roselle calyx on protein content (Hinojosa-Go´mez et al., 2018).

6.4 Utilization of Roselle protein products The use of different sources of protein can vary widely from one country to another depending on food habits and traditions of the community (Halimatul et al., 2007). Scattered information show that RS and leaves can be used as seasoning in curries, since they contain an acid with rhubarb-like flavor (Islam, 2019). Moreover, the seeds and leaves are rich in proteins and thus can be either ground into powder after roasting and then added in sauces and soups (Islam, 2019). Roselle products can be further processed and are critical sources of nutritive protein. These are derived from commonly consumed Roselle products such as roasted RS which constitute products such flour, boiled RS, and other consumable parts such as the calyces (Ismail et al., 2008). Roselle products may also be

References

used as substitute sources of individual amino acids like glutamic acid, leucine, aspartic acid, arginine, and lysine which are abundantly found in Roselle products, especially from seeds (Hainida et al., 2008; Ismail et al., 2008). Previous studies have revealed that adequate methionine and cysteine composition of RS are necessary for human nutritional requirement, and therefore, the role of Roselle as a source of these compounds is unequivocal. Roselle products can be utilized as function ingredients in food industry as well as in production of bioactive peptides because they are rich in protein contents (Elneairy, 2014; Tounkara et al., 2013a,b). Roselle flour form seeds or calyces can be used to increase the protein content in other food products. For example, Adedeji et al. (2015) observed that sorghum complementary foods containing Roselle and soybean flour had high protein content than that made up of sorghum flour alone.

6.5 Conclusion The various parts (seeds, flowers, calyces, and leaves) of the Roselle plant contain different amounts of proteins. The seeds have the highest protein content. Protein products from the Roselle plant are considered relatively low-cost alternative protein sources useful in dietary supplements or as ingredients for the food industry. It was also found that the choice of the processing method is crucial since the different methods have effects on the quality of proteins obtained. The type of processing also dictates the shelf life of the products. The protein products are utilized/consumed in a number of diverse and varied ways depending on technology, cultural, and economic status of the community.

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332. Duke, J. A. (1983). Hisbiscus sabdariffa L. Available online: http://www.hort.purdue.edu/ newcrop/duke_energy/hisbiscus_sabdariffa.html. Duke, J. A., & Atchley, A. A. (1984). Proximate analysis. In B. R. Christie (Ed.), The Handbook of Plant Science in Agriculture (pp. 427
434). Boca Raton, FL: CRC Press Inc.. Duwa, H., Oyawoye, E. O., & Njidda, A. A. (2012). Effect of processing methods on the utilization of sorrel seed meal by broilers. Pakistan Journal of Nutrition, 11(1), 38
46. Egbert, W. R., & Payne, C. T. (2009). Plant proteins. In R. Tarte´ (Ed.), Ingredients in Meat Products. New York, NY: Springer. Available from https://doi.org/10.1007/9780-387-71327-4_5. El-Adawy, T. A., & Khalil, A. H. (1994). Characteristic of Roselle seeds as a new source of protein and lipid. Journal of Agricultural and Food Chemistry, 42, 1896
1900. Elneairy, N. A. (2014). Comparative studies on Egyptian and Libyan Roselle seeds as a source of lipid and protein. Food and Nutrition Sciences, 5, 2237
2245. Garti, H., Garti, H. A., & Nyarko, G. (2019). Comparative studies on the nutrient composition of shade-dried and sun-dried leaves of Rosselle (Hibiscus sabdariffa L.). African Journal of Horticultural Science, 16, 29
42. Ghislain, M. T., Etengeneng, A. E., Sonia, M. N. H., Noelle, K. S. C., & Gouado, I. (2014). Proximate and mineral composition, protein quality of Hibiscus sabdariffa L. (Roselle) seeds cultivated in two agro ecological areas in Cameroon. International Journal of Nutrition and Food Sciences, 3(4), 251
258. Hainida, E. K. I., Amin, I., Normah, H., & Mohd.-Esa, N. (2008). Nutritional and amino acid contents of differently treated Roselle (Hibiscus sabdariffa L.) seeds. Food Chemistry, 111(4), 906
911. Halimatul, S. M. N., Amin, I., Mohd-Esa, N., Nawalyah, A. G., & Muskinah, M. (2007). Protein quality of Roselle (Hibiscus sabdariffa L.) seeds. ASEAN Food Journal, 14(2), 131
140. Hinojosa-Go´mez, J., Martin-Herna´ndez, C. S., Heredia, J. B., Leo´n-Fe´lix, J., Osuna-Enciso, T., & Muy-Rangel, M. D. (2018). Roselle (Hibiscus sabdariffa L.) cultivars calyx produced hydroponically: Physicochemical and nutritional quality. Chilean Journal of Agricultural Research, 78(4), 478
485. Hoffman, J. R., & Falvo, M. J. (2004). Protein
which is best? Journal of sports science & medicine, 3(3), 118. Islam, M. M. (2019). Food and medicinal values of Roselle (Hibiscus sabdariffa L. Linne Malvaceae) plant parts: A review. Open Journal of Nutrition and Food Sciences, 1(1), 1003. Ismail, A., Ikram, E. H. K., & Nazri, H. S. M. (2008). Roselle (Hibiscus sabdariffa L.) seeds nutritional composition protein quality and health benefits. Food, 2, 1
16. Jeannett, A. I., Diego, A. A., Manuel, S., Jose´, A. M., Nancy, V., Carlos, A. G., . . . Eduardo, M. (2020). Organic acids from Roselle (Hibiscus sabdariffa L.). A brief review of its pharmacological effects. Biomedicines, 8(5), 100. Available from https:// doi.org/10.3390/biomedicines8050100.

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Juhari, N. H., & Petersen, M. A. (2018). Physicochemical properties and oxidative storage stability of milled Roselle (Hibiscus sabdariffa L.) seeds. Molecules (Basel, Switzerland), 23, 385. Lakra, P., & Sehgal, S. (2011). Influence of processing on total and extractable mineral content of products prepared from potato flour. J. Food Sci. Technol., 48(6), 735
739. Mabrouk, O., Amin, W. A., Youssef, A. M., & Abou-Samaha, O. (2018). Roselle (Hibiscus Sabdariffa) Seeds and Kernels as a Potential Source of Oil, Protein and Minerals. Egyptian Journal of Food Science, 46, 55
67. Mahadevan, S., & Pradeep, K. (2009). Hibiscus sabdariffa Linn—An overview. Natural Product Radiance, 8, 77
83. Mariod, A. A., Mirghani, M. E. S., & Hussein, I. H. (2017). Unconventional Oilseeds and Oil Sources. London, UK: Academic Press. Mariod, A. A., Suryaputra, S., Hanafi, M., Rohmana, T., Kardono, L. B. S., & Herwan, T. (2013). Effect of different processing techniques on Indonesian Roselle (Hibiscus radiates) seed constituents. Acta Scientiarum Polonorum, Technologia Alimentaria, 12(4), 359
364. Mat Isa, A., Isa, P. M., & Abd Aziz, A. R. (1985). Analisis kimia dan pemprosesan Roselle (Hibiscus Sabdariffa L.). Mardi Research Bulletin, 13, 68
74. Messina, M. J. (1999). Legumes and soybeans: Overview of their nutritional pro-files and health effects. American Journal of Clinical Nutrition, 70, 39S
50S. Mokhtari, Z., Zarringhalami, S., & Ganjloo, A. (2018). Evaluation of chemical, nutritional and antioxidant characteristics of Roselle (Hibiscus sabdariffa L.) seed. Nutrition and Food Sciences Research, 5, 41
46. Moore, S., Spackman, D. H., & Stein, W. H. (1958). Chromatography of amino acids on sulfonated polystyrene resins. An improved system. Analytical Chemistry, 30(7), 1185
1190. Muhammed Amin, M., & Ali Kanimarani, M. S. S. (2020). The influence of plant growth regulators on phytochemical components in the leaves and calyxes of Roselle (Hibiscus sabdariffa L.). Zanco Journal of Pure and Applied Sciences, 32(3), 193
199. Available from https://doi.org/10.21271/ZJPAS.32.3.20. Naturland, e. V. (2000) Organic Farming in the Tropics and Subtropics Exemplary Description of 20 Crops. 1st ed. Germany; pp. 1a10. Nzikou, J. M., Bouanga-Kalou, G., Matos, L., Ganongo-Po, F. B., Mboungou-Mboussi, P. S., & Moutoula, F. E. (2011). Characteristics and nutritional evaluation of seed oil from Roselle (Hibiscus sabdariffa L.) in Congo-Brazzaville. Current Research Journal of Biological Sciences, 3(2), 141
146. Ojokoh, A. O. (2010). Fermentation studies on Roselle (Hibiscus sabdariffa) calyx neutralized with trona (sodium sesquicarbonate) at different concentrations. World Journal of Microbiology and Biotechnology, 26(12), 2257
2261. Petrusa´n, J., Jawel, H., & Huschek. (2016). Protein-rich vegetal sources and trends in human nutrition: A review. Current Topics in Peptide and Protein Research, 17, 1
18. Rimamcwe, K. B., Chavan, U. D., Dalvis, U. S., & Gaiwad, R. S. (2017). Nutritional quality of Roselle seed flour cookies. International Journal of Current Research, 9(12), 63053
63058. Salah, E. O., & Hayat, Z. E. (2009). Proximate composition of Karkadeh (Hibiscus sabdariffa) seeds and some functional properties of seed protein isolate as influenced by pH

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194. Available from https://doi.org/10.1080/09637480701664399. Schippers, R. R. (2000). African Indigenous vegetable: An overview of the cultivated species. Natural Resources Institute/ACP-EU. Technical Center for Agricultural and Rural Cooperation. Chatam, UK, pp. 1-124. Shaheen, A. M., El-Nakhlawy, S. F., & Al-Shareef, A. R. (2012). Roselle (Hibiscus sabdariffa L.) seeds as unconventional nutritional source. African Journal of Biotechnology, 11(41), 9821
9824. Singh, P., Khan, M., & Hailemariam, H. (2017). Nutritional and health importance of Hibiscus sabdariffa: A review and indication for research needs. Journal of Nutritional Health & Food Engineering, 6, 125
128. Available from https://doi.org/10.15406/ jnhfe.2017.06.00212. Teixeira, E. M., Carvalho, M. R., Neves, V. A., Silva, M. A., & Arantes-Pereira, L. (2014). Chemical characteristics and fractionation of proteins from Moringa oleifera Lam. leaves. Food Chemistry, 147, 51
54. Available from https://doi.org/10.1016/j. foodchem.2013.09.135. Tounkara, F., Amza, T., Lagnika, C., Le, G. W., & Shi, Y. H. (2013a). Extraction, characterization, nutritional and functional properties of Roselle (Hibiscus sabdariffa Linn) seed proteins. Songklanakarin Journal of Science & Technology, 35(2), 159
166. Tounkara, F., Bashari, M., Le, G., & Shi, Y. (2014). Antioxidant activities of Roselle (Hibiscus Sabdariffa L.) seed protein hydrolysate and its derived peptide fractions. International Journal of Food Properties, 17(9), 1998
2011. Tounkara, F., Sodio, B., Amza, T., Le, G., & Shi, Y. (2013b). Antioxidant effect and water-holding capacity of Roselle (Hibiscus sabdariffa L.) seed protein hydrolysates. Advance Journal of Food Science and Technology, 5(6), 752
757. Yacoub, A. A., & Abdalla, A. A. (2007). Effect of domestic processing methods on chemical, in vitro digestibility of protein and starch and functional properties of Bambara groundnut (Voandzeia subterranean) seed. Journal of Agriculture and Biological Sciences, 3, 24
34. Yagoub, A., & Mohammed, M. A. (2008). Fururndu, a meat substitute from fermented Roselle (Hibiscus sabdariffa L.) seed: Investigation on amino acids composition, protein fractions, minerals content and HCl-extractability and microbial growth. Pakistan Journal of Nutrition, 7, 352
358. Yagoub, A. A., Mohamed, E. B., Ahmed, A. H. R., & El Tinay, A. H. (2004). Study on Furundu, a traditional Sudanese Fermented Roselle (Hibiscus sabdariffa L.) seed: effect on in vitro protein digestibility, chemical composition, and functional properties of the total proteins. Journal of Agricultural and Food Chemistry, 52, 6143
6150. Young, R. V., & Pellet, L. P. (1994). Plant proteins in relation to human protein and amino acid nutrition. American Journal of Clinical Nutrition, 59, 1203S
1212S.

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Volatile compounds and phytochemicals of Hibiscus sabdariffa

7

Abubakr Musa1, Haroon Elrasheid Tahir2, Mohammed Abdalbasit A. Gasmalla3, Gustav Komla Mahunu4 and Abdalbasit Adam Mariod5,6 1

Sugar Institute, University of Gezira, Wad Madani, Sudan School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China 3 Department of Nutrition and Food Technology, Omdurman Islamic University, Omdurman, Sudan 4 Department of Food Science and Technology, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana 5 Indigenous Knowledge and Heritage Center, Ghibaish College of Science & Technology, Ghibaish, Sudan 6 College of Sciences and Arts-Alkamil, University of Jeddah, Alkamil, Saudi Arabia 2

Chapter Outline 7.1 Introduction ................................................................................................... 91 7.2 Bioactive constituents .................................................................................... 92 7.3 Organic acids ................................................................................................. 92 7.3.1 Ascorbic acid ................................................................................96 7.3.2 Hydroxycitric acid .........................................................................97 7.4 Anthocyanins ................................................................................................. 98 7.5 Flavonoids ..................................................................................................... 99 7.6 Volatile compounds ........................................................................................ 99 7.7 Conclusion ...................................................................................................107 References ..........................................................................................................108

7.1 Introduction In developing countries a large group of the population depends on plants and medicinal herbs in the treatment of primary health problems. The reasons that made traditional medicines more popular among most of the world’s population were cheap, abundant, and with a less harmful effect on health. Recently, many research plants worldwide have focused on exploring the tremendous potential of Roselle (Hibiscus sabdariffa). DOI: https://doi.org/10.1016/B978-0-12-822100-6.00015-X © 2021 Elsevier Inc. All rights reserved.

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plants used in various traditional medicinal systems. There are many medicinal plants that contain a large amount of phytochemicals, which have been used in the treatment of many diseases. The Roselle (Hibiscus sabdariffa) plant is one of the most important due to its health properties and benefits (Ali, Wabel, & Blunden, 2005). Roselle (Hibiscus sabdariffa) is considered one of the plants of high importance to humans because it contains many nutritional and therapeutic properties in most of its parts. Roselle plant is consumed in most parts of the world as a cold or hot drink. Roselle extracts are also used in folk medicine to treat many diseases, such as high blood pressure, liver disease, and fever (Ali, Wabel et al., 2005). There is a lot of research focused on the nutritional values and medicinal benefits of the hibiscus plant. To name a few, Singh, Khan, & Hailemariam (2017) discussed the nutritional and therapeutic benefits of Roselle. Morton (1987) and Luvonga, Njoroge, Makokha, and Ngunjiri (2010) have proven that the Roselle contains many vitamins such as niacin, riboflavin, pyridoxine, and ascorbic acid in significant proportions. And this supports the fact of the nutritional value of Roselle. Da-Costa-Rocha, Bonnlaender, Sievers, Pischel, and Heinrich (2014) reported the phytochemical and pharmacological properties of Roselle. The containment of Roselle for phytochemicals was widely confirmed a long time ago, and many of these active compounds are still discovered. These were helped by the spread of modern devices such as Ultra-High-Performance Liquid Chromatography and Liquid Chromatography-Mass Spectroscopy. In this chapter, most of the available published information about the minor constituents and phytochemicals of Roselle will be discussed.

7.2 Bioactive constituents Previous studies have shown that the extracts of the Roselle plant have antioxidant, anticholesterol, and antidiabetic effects due to their high levels of bioactive compounds (Da-Costa-Rocha, Bonnlaender et al., 2014) such as organic acid (hydroxycitric acid and hibiscus acid), anthocyanins (delphinidin
3-sambubioside and cyanidine-3-sambubioside) (Fernndez-Arroyo, Rodrı´guez-Medina et al., 2011), phenolic acids (protocaticoic acid and caffeic acid (Kuo, Kao et al., 2012), flavonoids (epigallocatechin gallate and catechin (Kuo, Kao et al., 2012), and volatile compounds (3-methylbutan-1-ol, phenylacetaldehyde, and acetic acid (AvalosMartı´nez, Pino, Sa´yago-Ayerdi, Sosa-Moguel, & Cuevas-Glory, 2019). Table 7.1 reveals an overview of the phytochemical components in Roselle calyx detected on liquid chromatography and gas chromatography.

7.3 Organic acids The primary organic acids present in Roselle were hydroxycitric acid, hibiscus acid, citric acid, malic acid, and tartaric acids, and ascorbic acid, succinic acid,

7.3 Organic acids

Table 7.1 Phytochemical compounds of Roselle (Hibiscus sabdariffa) calyces. Classes

Compounds

References

Phenolic acids

Chlorogenic acid isomer I

Fernndez-Arroyo, Rodríguez-Medina et al. (2011) Fernnndez-Arroyo, Rodríguez-Medina et al. (2011), Jamini, Islam et al. (2019) Fernndez-Arroyo, Rodríguez-Medina et al. (2011) Sinela, Rawat et al. (2017), Cassol, Rodrigues, and Zapata Noreña (2019), Piovesana and Noreña (2019) Sinela, Rawat et al. (2017), Cassol, Rodrigues et al. (2019), Piovesana and Noreña (2019) Escobar-Ortiz, Castaño-Tostado, RochaGuzmán, Gallegos-Infante, and ReynosoCamacho (2020) Sinela, Rawat et al. (2017), Cassol, Rodrigues et al. (2019), Piovesana and Noreña (2019), Escobar-Ortiz, CastañoTostado et al. (2020) Jabeur, Pereira et al. (2017) Kuo, Kao et al. (2012), Ifie, Ifie et al. (2018), Escobar-Ortiz, Castaño-Tostado et al. (2020) Escobar-Ortiz, Castaño-Tostado et al. (2020) Kuo, Kao et al. (2012), Ifie, Ifie et al. (2018), Escobar-Ortiz, Castaño-Tostado et al. (2020) Fernndez-Arroyo, Rodríguez-Medina et al. (2011) Fernndez-Arroyo, Rodríguez-Medina et al. (2011), Jamini, Islam et al. (2019) Jamini, Islam et al. (2019) Jamini, Islam et al. (2019), PimentelMoral, Borrás-Linares et al. (2019) Jamini, Islam et al. (2019), PimentelMoral, Borrás-Linares et al. (2019) Jamini, Islam et al. (2019) Jamini, Islam et al. (2019), PimentelMoral, Borrás-Linares et al. (2019) Jamini, Islam et al. (2019) Jamini, Islam et al. (2019)

Chlorogenic acid Chlorogenic acid isomer II 3-Caffeoylquinic acid

5-Caffeoylquinic acid

Quinic acid

4-Caffeoylquinic acid

5-O-Caffeoylquinic acid Caffeic acid

Coumaric acid Protocatechuic acid

Hydroxycitric acid Hibiscus acid Hydroxicitric acid Neochlorogenic acid Cryptochlorogenic acid Methyl digallate Methyl chlorogenate Coumaroilquinic acid Dihydroferulic

(Continued)

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Table 7.1 Phytochemical compounds of Roselle (Hibiscus sabdariffa) calyces. Continued Classes

Compounds

References

5-O-caffeoylshikimic acid

Fernndez-Arroyo, Rodríguez-Medina et al. (2011), Cassol, Rodrigues et al. (2019), Jamini, Islam et al. (2019), Pimentel-Moral, Borrás-Linares et al. (2019), Piovesana and Noreña (2019) Pimentel-Moral, Borrás-Linares et al. (2019), Escobar-Ortiz, Castaño-Tostado et al. (2020) Jamini, Islam et al. (2019), PimentelMoral, Borrás-Linares et al. (2019) Jamini, Islam et al. (2019), PimentelMoral, Borrás-Linares et al. (2019) Jabeur, Pereira et al. (2017) Pimentel-Moral, Borrás-Linares et al. (2019) Ifie, Ifie et al. (2018), Escobar-Ortiz, Castaño-Tostado et al. (2020) Jabeur, Pereira et al. (2017), Ifie, Ifie et al. (2018) Piovesana and Noreña (2019) Cassol, Rodrigues et al. (2019), Piovesana and Noreña (2019) Cassol, Rodrigues et al. (2019) Escobar-Ortiz, Castaño-Tostado et al. (2020) Escobar-Ortiz, Castaño-Tostado et al. (2020) Escobar-Ortiz, Castaño-Tostado et al. (2020) Pimentel-Moral, Borrás-Linares et al. (2019) Pimentel-Moral, Borrás-Linares et al. (2019) Kuo, Kao et al. (2012) Kuo, Kao et al. (2012) Fernndez-Arroyo, Rodríguez-Medina et al. (2011), Jamini, Islam et al. (2019), Pimentel-Moral, Borrás-Linares et al. (2019) Fernndez-Arroyo, Rodríguez-Medina et al. (2011), Cassol, Rodrigues et al. (2019) Fernndez-Arroyo, Rodríguez-Medina et al. (2011), Cassol, Rodrigues et al. (2019), Piovesana and Noreña (2019)

Chlorogenic acid

Ethylchlorogenate Ethylchlorogenate isomer II 5-(Hydroxymethyl)furfural Ethylchlorogenate isomer III Gallic acid 3-O-caffeoylquinic acid Ferulic acid-derived 5-p-Coumaroylquinic acid 3-p-Coumaroylquinic acid 2,5-Dihydroxybenzoic acid Syringic acid Ellagic acid Coumaroylquinic acid

Flavonoids

Dihydroferulic acid-4Oglucuronide Epigallocatechin gallate Catechin Quercetin

Quercetin-3-sambubioside

Quercitin-3-glucoside

(Continued)

7.3 Organic acids

Table 7.1 Phytochemical compounds of Roselle (Hibiscus sabdariffa) calyces. Continued Classes

Compounds

References

Kaempferol-3-O-rutinoside

Fernndez-Arroyo, Rodríguez-Medina et al. (2011), Piovesana and Noreña (2019) Fernndez-Arroyo, Rodríguez-Medina et al. (2011), Piovesana and Noreña (2019) Fernndez-Arroyo, Rodríguez-Medina et al. (2011), Jabeur, Pereira et al. (2017) Jamini, Islam et al. (2019)

Kaempferol-3-pCoumarylglucoside Myricetin-pentosylhexoside Myricetin-3arabinogalactoside Quercetin-3-sambubioside Quercetin-3-rutinoside Quercetin-3-glucoside Kaempferol-3-O-rutinoside Methylepigallocatechin Myricetin Kaempferol Myricetin 3-Oarabinogalactoside Myricetin 3-sambubioside Quercetin 3-Osambubioside Quercetin-3-rutinoside Anthocyanins

Cyanidine-3-sambubioside

Prodelphinidin B3 Delphinidin-3-Osambubioside Delphinidin-3-O-glucoside

Cyanidin 3-O-sambubioside

Cyanidin-3-O-glucoside Petunidin-3-O-glucoside

Jamini, Islam et al. (2019) Fernndez-Arroyo, Rodríguez-Medina et al. (2011), Jamini, Islam et al. (2019) Jamini, Islam et al. (2019) Cassol, Rodrigues et al. (2019), Jamini, Islam et al. (2019) Jamini, Islam et al. (2019), PimentelMoral, Borrás-Linares et al. (2019) Jamini, Islam et al. (2019), PimentelMoral, Borrás-Linares et al. (2019) Jamini, Islam et al. (2019), PimentelMoral, Borrás-Linares et al. (2019) Ifie, Ifie et al. (2018) Cassol, Rodrigues et al. (2019), Piovesana and Noreña (2019) Ifie, Ifie et al. (2018), Piovesana and Noreña (2019) Cassol, Rodrigues et al. (2019), Piovesana and Noreña (2019) Fernndez-Arroyo, Rodríguez-Medina et al. (2011), Kouakou, Konkon et al. (2015), Jabeur, Pereira et al. (2017) Jamini, Islam et al. (2019) Kouakou, Konkon et al. (2015), Ifie, Ifie et al. (2018) Kouakou, Konkon et al. (2015), Ifie, Ifie et al. (2018), Cassol, Rodrigues et al. (2019) Kouakou, Konkon et al. (2015), Ifie, Ifie et al. (2018), Cassol, Rodrigues et al. (2019) Kouakou, Konkon et al. (2015) Kouakou, Konkon et al. (2015) (Continued)

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CHAPTER 7 Volatile compounds and phytochemicals of Roselle

Table 7.1 Phytochemical compounds of Roselle (Hibiscus sabdariffa) calyces. Continued Classes

Compounds

References

Malvidin-3-O-glucoside

Kouakou, Konkon et al. (2015)

N-Feruloyltiramine 2,4,6Trihydroxybenzaldehyde N-Feruloyltyramine

Jamini, Islam et al. (2019) Escobar-Ortiz, Castaño-Tostado et al. (2020) Pimentel-Moral, Borrás-Linares et al. (2019)

Others

and oxalic acid were in less amount. Generally, the percentage of organic acids in Hibiscus sabdariffa is very close, as it found that the hibiscus acid is the most representative acid of Hibiscus species (13%
24%), whereas the other organic acids range from 12% to 20% for citric acid, 2%
9% for malic acid, 8% for tartaric acid, and 0.02%
0.05% for ascorbic (Riaz & Chopra, 2018). At the end of 1930s, malic acid, citric acid, and tartaric acid were detected for the first time in Roselle infusions (Buogo & Picchinenna, 1937; Indovina and Capotummino, 1938); however, tartaric acid was detected only as a trace component by Indovina and Capotummino (1938). In 1980 also, citric and malic acids were reported in five strains of Roselle from different geographical origins (from Egypt, Senegal, India, Thailand, and Central America). In 2018 Ifie, Ifie, Ibitoye, Marshall, and Williamson (2018) investigated the effect of wet and dry seasons on oxalic acid, tartaric acid, malic acid, citric acid, and succinic acid Roselle calyces grown in South-Western Nigeria. When the dry compared with rainy season malic acid was significantly higher value (45.6 mg/100 g dry sample), tartaric acid showed a lower value (0.11 mg/100 g dry sample), whereas oxalic acid was reported as a trace value (0.060 mg/100 g dry samples). Jabeur et al. (2017) chemically characterized the Roselle extracts, reporting the existence of a variety of organic acids such as oxalic acid, malic acid, shikimic acid, and fumaric acid with malic acid being the major compound (9.10 g/100 g dw), whereas fumaric acid showed a lower value (0.043 g/100 g dw). Malic acid has been described to have health benefits, specifically, its ability to diminish the risk of me toxicity, privilege fluidity, aid the maintenance of oral health and increase immunity (Hossain, Akhtar, & Anwar, 2015). Thus Roselle plants could be industrially used for the production of malic acid, and it is derivatives due to their functional properties (Jabeur, Pereira et al., 2017; Lin-Holderer, Li, Gruneberg, Marti, & Kunze, 2016).

7.3.1 Ascorbic acid Ascorbic acid is also reported in aqueous extract of Roselle, and it is concentration differs according to the variety, season, harvest conditions, genetics and geographical

7.3 Organic acids

origins (Da-Costa-Rocha, Bonnlaender et al., 2014). Ascorbic acid in Nigerian white and red and deep red Roselle was 86.5 mg/100 g, 63 mg/100 g, and 54 mg/100 g, respectively (Babalola, Babalola, & Aworh, 2001). While for Sudanese white and red Roselle were 11.0 mg/100 g and 15.5 mg/100 g, respectively (Tahir, Xiaobo et al., 2017). The concentration of ascorbic acid in the Malaysian red Roselle was significantly higher than once previously reported in Nigerian and Sudanese samples (Ismail, Ikram, & Nazri, 2008). The concentrations of ascorbic acid of 35 available genotypes of Roselle collected from Bangladesh were ranged between 2.62 and 42.42 mg/100 g (Jamini, Islam, Mohi-ud-Din, & Saikat, 2019).

7.3.2 Hydroxycitric acid Hydroxycitric acid (Fig. 7.1) is citric acid with an additional hydroxyl group at carbon number two. It consists of lactone forms, and four stereoisomers, (2S, 3S), (2R, 3R), (2S,3R), and (2R, 3S). The (2S,3R)-hydroxycitric acid consider as the main organic acid found in the Roselle calyces (Hida, Yamada, & Yamada, 2007). It is worth observing that (2S,3R)-hydroxy acid of Roselle is different from the most known (2S,3S)-hdroxy citric acid obtained from other sources such as Garcinia sp. Therefore it is an important question to understand whether both diastereomers (2S, 3R) and (2S, 3S) are identical or partially different in pharmacological profiles (Da-Costa-Rocha, Bonnlaender et al., 2014).

7.3.2.1 Hibiscus acid Hibiscus acid (Fig. 7.2) consider as one of the main organic acids found in the extracts of the Roselle calyces and leaves (Herranz-Lpez, Ferna´ndez-Arroyo et al., 2012; Peng, Chyau et al., 2011). Hibiscus acid contains in its structure a citric acid fraction with an additional hydroxyl group in the second carbon and two diastereomers because of the presence of two chiral centers in the molecule (Boll, 1969; Griebel, 1939). Hibiscus acid is found in Roselle plant flowers in

FIGURE 7.1 (2S,3R)-Hydroxycitric acid (Da-Costa-Rocha, Bonnlaender et al., 2014).

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CHAPTER 7 Volatile compounds and phytochemicals of Roselle

FIGURE 7.2 Hibiscus acid (Da-Costa-Rocha, Bonnlaender et al., 2014).

FIGURE 7.3 Chemical structures of main anthocyanins (Da-Costa-Rocha, Bonnlaender et al., 2014).

rates ranging from 13% to 24% (Eggensperger & Wilker, 1996). This variation in proportions may be due to reasons related to different strains, genes, and environmental conditions.

7.4 Anthocyanins The anthocyanins are natural pigments present in the dried flowers of the Roselle plant. The anthocyanins are derivatives of flavonoids. From the pigment of Roselle calyces, there are three types of anthocyanins identified, including hibiscin (delphinidin-3-sambubioside), chrysanthin (cyanidine-3-glucoside), and delphinidin-3-glucoside (Fig. 7.3) (Du & Francis, 1973). There is a study done with five different strains of Roselle plants. Reported that the main compounds found in the Roselle plant are cyanidine 3-sambiobioside and cyanidine-3 glucose (Khafaga, Koch, El Afry, & Prinz, 1980). Also, several studies determined that the main anthocyanins found in extracts of Roselle calyces and leaves are delphinidin3-sambubioside. (Delphinidin-3-O-(2-O-b-D-xylopyranosyl)-b-D-glucopyranoise) and cyanidine
3-sambubioside (cyanide 3-O-(2-O-B-D-xylopyranosyl)-b-D-glucopyranoside) (Alarcon-Aguilar, Zamilpa et al., 2007; Herranz-Lpez, Ferna´ndez-Arroyo et al., 2012; Peng, Chyau et al., 2011; Ramrezaˆ Rodrigues, Balaban, Marshall, & Rouseff, 2011; Rodrı´guez-Medina, Beltra´n-Debo´n et al., 2009).

7.6 Volatile compounds

7.5 Flavonoids Numerous scientific research has revealed that Roselle calyces are rich in flavonoids and polyphenols, which have enhanced the nutritional value of Roselle, as their antioxidant properties distinguish these compounds. Polyphenols of the flavonol type and flavanol found in Roselle are in the form of a simple or polymer form. The most flavonoids isolated from the Roselle extracts were, hibiscetin3-glucoside (hibiscitrin), gossypitrin, gossytrin, sabdaritrin and other gossypetin glucosides, quercetin and luteolin; also, chlorogenic acid, protocatechuic acid, pelargonidic acid, eugenol, quercetin, luteolin, and the sterols b-sitosterol and ergosterol (McKay, 2009; Williamson, Driver, & Baxter, 2009).

7.6 Volatile compounds The odor of Roselle is due to the presence of volatile compounds. A study conducted in 1992 demonstrated that the Roselle plant contains more than 25 volatile compounds (Jirovetz, Jaeger et al., 1992). Later, about 37 volatile compounds in Roselle extracts were identified, and these compounds including, sugar derivatives, fatty acid derivatives, terpenes, phenolic derivatives, and miscellaneous compounds (Chen, Huang, Ho, & Tsai, 1998). In other research, the volatile compounds profile of Roselle calyces water extracts under different extraction conditions was examined by GC-MS. About 32 compounds were identified and divided into five chemical groups: aldehydes, alcohols, ketones, terpenes, and acids (Ramrezaˆ Rodrigues, Balaban et al., 2011). Seven of the common aroma compounds identified in these extracts are hexanal, 3-octanone, octanal, 1-octen-3one, nonanal, 2,4-nonadienal(E, E), and geranylacetone. Tahir et al. (2017) studied the aroma profiles of red and white Roselle collected from four locations in Sudan (Khartoum, Al-Rahad, Al-Fashir, and Al-Gezira). In total 19 volatile compounds have been identified in Roselle extracts, including aldehydes and alcohols, ketones, esters, and phenols. Significant differences in the content of these volatile compounds were detected in samples collected in each location, indicating that the samples varied in aroma quality. Al-Rahad sample was characterized by the highest esters followed by Al-Fashir and Al-Gezira type, but the white Roselle revealed the lowest value. The authors contributed the highest pleasant aroma of Al-Rahad samples to higher ester compounds content. (Ramı´rezRodrigues, Plaza, Azeredo, Balaban, & Marshall, 2012) investigated the effect of dense phase carbon dioxide (DPCD) on the volatile profile of Hibiscus sabdariffa beverage during storage. Samples treated with the DPCD technique were compared with the conventional thermal method and untreated samples (control) during cold (4 C) storage. The results showed that DPCD practice retained more volatile compounds as compared to thermally processed beverages. The volatile compounds of the beverages consisted of alcohols and aldehydes with 1-octen-3-ol, decanal,

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octanal, 1-hexanol, and nonanal as the volatiles with the maximum relative percentage peak areas. However, the volatile intensities were not evaluated and should be measured in future works. Camelo-Me´ndez et al. (2013) evaluated the volatile profiles of four varieties (Negra, Sudan, Rosa, and Blanca Roselle) of Mexican Roselle. The samples were extracted using water and ethanol. The ethanolic extract of Rosa variety obtained more volatile compounds compared to other varieties. The aqueous extract of Rosa showed high concentrations of ethanol, geraniol, menthol, 2-nonanol, benzaldehyde, linalool, gamma undecalactone, and ethyl methyl phenylglycidate as compared to other varieties. Generally, geraniol was the key volatile in extracts from the four varieties. Juhari, Varming, and Petersen (2015) evaluated volatile compounds of Roselle dynamic headspace sampling using different sample preparation techniques. In this study a total of 125 compounds were detected comprising terpenes (32), aldehydes (20), esters (16), ketones (14), alcohols and furans (13), acids (9), sulfurs (3), lactones (2), and others (3). Roselle quality is strongly affected by many determinants, and the geographical origin is one of the main factors (Tahir, Arslan et al., 2020). In a study, Juhari, Bredie, Toldam-Andersen, and Petersen (2018) studied the different volatile compounds of Roselle calyces collected from different geographical origins. Seventeen samples of Roselle calyces were obtained from eight countries, namely Australia, China, Chad, Malaysia, Mexico, Nigeria, Sudan, and Thailand. A total of 135 compounds were detected including terpenes, aldehydes, esters, furans, and ketones. Among the volatiles identified in 17 samples, α-terpinolene, hexanal, butanal, 2-hexenal, 2-methyl-1-penten-3-one, 1-octen-3-one, 2,6,6-trimethylcyclohexanone, 2-methylfuran, 5-isoprenyl-2methyl-2-vinyltetrahydrofuran, herboxide second isomer, and 1-octen-3-ol were the key odorants identified consistently in gas chromatography-olfactometry analysis. This study produced important information for future commercial usages of Roselle in the food industry. Zannou, Kelebek, and Selli (2020) investigated the odorants in Roselle infusions prepared by hot and cold methods. Three infusions were prepared as follow: (1) 2.5% (wt./vol.) of Roselle calyces powder was brewed at 98 C for 16 minutes (R16m); (2) 5% (wt./vol.) was brewed at 98 C for 40 minutes (R40M); and (3) 5% (wt./vol.) was brewed at 25 C for 24 hours (R24H). A total of 38, 38, and 39 volatile compounds were detected in R16M, R40M, and R24H infusions, respectively. The identified compounds include alcohols, furans, acids, ketones, aldehydes, phenols, lactones, pyranone, pyrrole, terpene, and ester. R24H infusion showed a higher total aroma concentration as compared to other treatments. In all infusions, furans were found to be the major volatile class followed by alcohols. According to the results of the volatile compounds extract dilution analysis, a total of 22 and 23 different key volatile compounds were identified in hot extraction and cold extraction, respectively. Also, analysis of the aroma-active compound indicated that 15 of the 19 were reported for the first time in Roselle infusions. Based on the above finding, it is clear that the extraction methods have an important effect on the final aroma and key odorants of Roselle infusions.

7.6 Volatile compounds

Farag, Rasheed, and Kamal (2015) investigated the volatile compounds of Roselle collected from Sudan and Egypt. A total of 104 compounds were identified of which only 32 were previously detected in Roselle infusions. Aldehydes and furans were the major compounds detected in both cultivars. The main difference in aroma profiles was the higher concentration of acetic acid and furfural in Sudanese samples, whereas 1-octen-3-ol was abundant in Egyptian Roselle. Gonzalez-Palomares, Estarro´n-Espinosa, Go´mez-Leyva, and AndradeGonza´lez (2009) investigated the effect of various temperatures (150, 160, 170, 180, 190, 200, and 210 C) on the aroma profile of Roselle infusions. In total 20 aroma compounds were detected in the extracts including terpenoids, esters, hydrocarbons, and aldehydes. Among the volatiles identified, 14 of them were identified in the powder, whereas 10 compounds were detected in Roselle extract, indicating the loss of some components due to the degradation process. Roselle extracts dried at 190 C showed the highest content of aroma compounds and the least concentrations of degraded compounds compared with other spray drying temperatures. The authors concluded that it is a challenge to produce Roselle powder exactly similar to the aqueous extract. Similar variations were observed when Ndiaye (2016) investigated the volatile profiles of the cold and hot extracts and their instant powders. Table 7.2 reveals a summary of the volatile components identified in aqueous extracts of Roselle.

Table 7.2 Volatile constituents of Roselle (Hibiscus sabdariffa) calyces. Classes

Compounds

References

Aldehydes

Phenylacetaldehyde (E)-Cinnamaldehyde Decanal

Avalos-Martínez, Pino et al. (2019) Avalos-Martínez, Pino et al. (2019) Pino, Márquez, and Marbot (2006), Gonzalez-Palomares, Estarrón-Espinosa et al. (2009) Chen, Huang et al. (1998), GonzalezPalomares, Estarrón-Espinosa et al. (2009), Ramírez-Rodrigues, Plaza et al. (2012) Chen, Huang et al. (1998) Chen, Huang et al. (1998), Tahir, Xiaobo et al. (2017) Chen, Huang et al. (1998), Pino, Márquez et al. (2006) Chen, Huang et al. (1998), Pino, Márquez et al. (2006) Chen, Huang et al. (1998), Pino, Márquez et al. (2006) Chen, Huang et al. (1998) Zannou, Kelebek et al. (2020)

Furfural

2,2-Dimethylhexanal Hexanal (E)-2-Hexenal Heptanal (E)-2-Heptenal 5-Methyl-2-furaldehyde 5-Hydroxymethylfurfural

(Continued)

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CHAPTER 7 Volatile compounds and phytochemicals of Roselle

Table 7.2 Volatile constituents of Roselle (Hibiscus sabdariffa) calyces. Continued Classes

Compounds

References

Nonanal

Ramírez-Rodrigues, Plaza et al. (2012), Tahir, Xiaobo et al. (2017) Ramírez-Rodrigues, Plaza et al. (2012) Chen, Huang et al. (1998), RamírezRodrigues, Plaza et al. (2012), AvalosMartínez, Pino et al. (2019) Ramírez-Rodrigues, Plaza et al. (2012), Tahir, Xiaobo et al. (2017) Tahir, Xiaobo et al. (2017)

Decanal Octanal

Benzaldehyde

Phenol derivatives

2-Furancarboxaldehyde, 5-methylSafranal 2-Phenyl-2-butenal 5-Methyl-2-furfural (E)-2-Nonenal 2-Methylbutanal p-Menthen-9-al Undecanal (E,E)-2,4-Decadienal (E)-Isoeugenol (Z)-Phyto Eugenol Phenol, 2,4-di-tert-butyl 4-Vinylguaiacol 4-Vinylphenol 2-Methoxy-4-vinylphenol

Acids

4-Vinylguaiacol Methyl eugenol Acetic acid

Propanoic acid 2-Methylbutanoic acid Pentanoic acid Hexanoic acid (Z)-3-Hexenoic acid Heptanoic acid

Tahir, Xiaobo et al. (2017) Tahir, Xiaobo et al. (2017) Zannou, Kelebek et al. (2020) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Avalos-Martínez, Pino et al. (2019) Avalos-Martínez, Pino et al. (2019) Chen, Huang et al. (1998), GonzalezPalomares, Estarrón-Espinosa et al. (2009) Tahir, Xiaobo et al. (2017) Zannou, Kelebek et al. (2020) Zannou, Kelebek et al. (2020) Pino, Márquez et al. (2006), AvalosMartínez, Pino et al. (2019) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Chen, Huang et al. (1998), RamírezRodrigues, Plaza et al. (2012), AvalosMartínez, Pino et al. (2019) Avalos-Martínez, Pino et al. (2019) Pino, Márquez et al. (2006), AvalosMartínez, Pino et al. (2019) Avalos-Martínez, Pino et al. (2019) Avalos-Martínez, Pino et al. (2019), Zannou, Kelebek et al. (2020) Avalos-Martínez, Pino et al. (2019) Avalos-Martínez, Pino et al. (2019) (Continued)

7.6 Volatile compounds

Table 7.2 Volatile constituents of Roselle (Hibiscus sabdariffa) calyces. Continued Classes

Compounds

References

2-Ethylhexanoic acid Octanoic acid

Avalos-Martínez, Pino et al. (2019) Avalos-Martínez, Pino et al. (2019), Zannou, Kelebek et al. (2020) Avalos-Martínez, Pino et al. (2019), Escobar-Ortiz, Castaño-Tostado et al. (2020) Escobar-Ortiz, Castaño-Tostado et al. (2020) Avalos-Martínez, Pino et al. (2019) Avalos-Martínez, Pino et al. (2019) Pino, Márquez et al. (2006), AvalosMartínez, Pino et al. (2019) Zannou, Kelebek et al. (2020) Zannou, Kelebek et al. (2020) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Avalos-Martínez, Pino et al. (2019) Avalos-Martínez, Pino et al. (2019) Avalos-Martínez, Pino et al. (2019) Avalos-Martínez, Pino et al. (2019) Avalos-Martínez, Pino et al. (2019) Avalos-Martínez, Pino et al. (2019) Avalos-Martínez, Pino et al. (2019) Avalos-Martínez, Pino et al. (2019) Avalos-Martínez, Pino et al. (2019)

Benzoic acid

Salicylic acid Decanoic acid Tetradecanoic acid Hexadecanoic acid

Ketones

2-Methyl butanoic acid Nonanoic acid Isobutanoic acid Dodecanoic acid Tetradecanoic acid Pentadecanoic acid (Z)-9-Hexadecenoic acid 2,3-Butanedione 2-Butanone 1-Penten-3-one 2,3-Pentanedione 3-Hydroxybutan-2-one 1-Hexen-3-one Hexan-3-one 4-Methyl-3-penten-2-one 2-Methyltetrahydrofuran3-one 5-Methyl-2(3H)-furanone Heptan-2-one 1-Octen-3-one 2,4-Dihydroxy-2,5dimethyl-3(2H)-furanone 2,2,6Trimethylcyclohexan-1one (E)-6-Methyl-3,5heptadien-2-one a-Ionone Geranylacetone

Avalos-Martínez, Avalos-Martínez, Avalos-Martínez, Avalos-Martínez,

Pino et Pino et Pino et Pino et

al. al. al. al.

(2019) (2019) (2019) (2019)

Avalos-Martínez, Pino et al. (2019)

Avalos-Martínez, Pino et al. (2019) Avalos-Martínez, Pino et al. (2019) Avalos-Martínez, Pino et al. (2019) (Continued)

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CHAPTER 7 Volatile compounds and phytochemicals of Roselle

Table 7.2 Volatile constituents of Roselle (Hibiscus sabdariffa) calyces. Continued Classes

Compounds

References

Benzophenone Hexahydrofarnesyl acetone (E,E)-Farnesyl acetone 3-Octanone Hepten-2-one 6-Methyl-5-hepten-2-one

Avalos-Martínez, Pino et al. (2019) Avalos-Martínez, Pino et al. (2019)

Ethanone,1-(2-methyl-1cyclopenten-1-yl) Geranyl acetone 3-Methyl-3-buten-2-one 3-Hydroxy-2-butanone 2-Octanone 1-Hydroxy-2-propanone 2-Pentanone Geranyl acetone 6,10,14-Trimethyl-2pentadecanone γ-Undecalactone

Alcohols

5-Hepten-2-one, 6methylEhyl alcohol (Z)-3-Hexenol 2-Hexenol 1-Hexanol 1-Ocatnol 1-Octen-3-ol 1-Hexanol, 2-ethyl1-Nonanol Phenylethyl alcohol 2-Methyl-3-buten-2-ol 3-Penten-2-ol 2-(2-Butoxyethoxy) ethanol 2-Phenoxy ethanol

Avalos-Martínez, Pino et al. (2019) Ramírez-Rodrigues, Plaza et al. (2012) Ramírez-Rodrigues, Plaza et al. (2012) Pino, Márquez et al. (2006), RamírezRodrigues, Plaza et al. (2012) Tahir, Xiaobo et al. (2017) Tahir, Xiaobo et al. (2017) Zannou, Kelebek et al. (2020) Zannou, Kelebek et al. (2020) Zannou, Kelebek et al. (2020) Zannou, Kelebek et al. (2020) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Camelo-Méndez, Ragazzo-Sánchez et al. (2013) Tahir, Xiaobo et al. (2017) Camelo-Méndez, Ragazzo-Sánchez et al. (2013) Chen, Huang et al. (1998), Zannou, Kelebek et al. (2020) Chen, Huang et al. (1998) Chen, Huang et al. (1998), RamírezRodrigues, Plaza et al. (2012) Ramírez-Rodrigues, Plaza et al. (2012) Ramírez-Rodrigues, Plaza et al. (2012) Tahir, Xiaobo et al. (2017) Tahir, Xiaobo et al. (2017) Tahir, Xiaobo et al. (2017), Zannou, Kelebek et al. (2020) Zannou, Kelebek et al. (2020) Zannou, Kelebek et al. (2020) Zannou, Kelebek et al. (2020) Zannou, Kelebek et al. (2020) (Continued)

7.6 Volatile compounds

Table 7.2 Volatile constituents of Roselle (Hibiscus sabdariffa) calyces. Continued Classes

Compounds

References

Terpenes

Furfuryl alcohol Benzyl alcohol Isophytol 3-Methyl-1-butanol 2-Methyl-1-butanol 2-Furfuryl alcohol (Z)-3-Hexan-1ol (E)-Nerolidol Decanol p-Cymene

Zannou, Kelebek et al. (2020) Zannou, Kelebek et al. (2020) Zannou, Kelebek et al. (2020) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Gonzalez-Palomares, Estarrón-Espinosa et al. (2009), Avalos-Martínez, Pino et al. (2019) Gonzalez-Palomares, Estarrón-Espinosa et al. (2009), Avalos-Martínez, Pino et al. (2019) Chen, Huang et al. (1998) Chen, Huang et al. (1998) Chen, Huang et al. (1998) Chen, Huang et al. (1998) Gonzalez-Palomares, Estarrón-Espinosa et al. (2009) Gonzalez-Palomares, Estarrón-Espinosa et al. (2009), Camelo-Méndez, RagazzoSánchez et al. (2013) Pino, Márquez et al. (2006) Ramírez-Rodrigues, Plaza et al. (2012) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006), CameloMéndez, Ragazzo-Sánchez et al. (2013) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Chen, Huang et al. (1998) Chen, Huang et al. (1998)

Limonene

1,8-Cineole 1,4-Cineole Terpinolene p-Cymenene Alpha-Terpinolene Linalool

Linalool oxide Dehydroxylinalool oxide Myrcenol Terpinen-4-ol α-Terpinene α-Terpineol Cis-β-terpineol Geraniol β-Caryophyllene α-Humulene α-Calacorene β-Santalene (E)-Phytol R-Terpineol R-Farnesene

(Continued)

105

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CHAPTER 7 Volatile compounds and phytochemicals of Roselle

Table 7.2 Volatile constituents of Roselle (Hibiscus sabdariffa) calyces. Continued Classes

Hydrocarbons

Compounds

References

2-Methyl-6-methylene-7octen-2-ol 2-Ethenyltetrahydro2,6,6-trimethyl-2H-pyran P-Xylene P-Cymene (z)-β-Ocimene P-Cymene-8-ol 1,2-Dihydro-2,5,8trimethylnaphthalene 2,6-Dimethyl naphthaleneb 1-Methylnaphthalene

Pino, Márquez et al. (2006)

Furanic linalool oxide Z and E 2-Ethylfuran

Gonzalez-Palomares, Estarrón-Espinosa et al. (2009) Chen, Huang et al. (1998), Zannou, Kelebek et al. (2020) Chen, Huang et al. (1998) Chen, Huang et al. (1998)

Chen, Huang et al. (1998) Pino, Pino, Pino, Pino, Pino,

Márquez Márquez Márquez Márquez Márquez

et et et et et

al. al. al. al. al.

(2006) (2006) (2006) (2006) (2006)

Pino, Márquez et al. (2006) Pino, Márquez et al. (2006)

Furans

2-Pentylfuran Tetrahydro-2,2-dimethyl5-(1-methylpropyl)furan 2-Acetylfuran

Esters

2-Acetyl-5-methylfuran 2(5H)-Furanone Furfural acetone 2,2,6-Trimethyl-6vinyltetrahydrofuran Ethyl linoleate Ethyl linoleolate Ethyl acetate Ethyl benzoateb 2-Methylpropyl acetate 1-Butanol, 3-methyl-, acetate 4-Methyl-1-(1methylethyl)-3cyclohexen-1-ol acetate

Pino, Márquez et al. (2006), Zannou, Kelebek et al. (2020) Zannou, Kelebek et al. (2020) Zannou, Kelebek et al. (2020) Zannou, Kelebek et al. (2020) Pino, Márquez et al. (2006) Gonzalez-Palomares, Estarrón-Espinosa et al. (2009) Gonzalez-Palomares, Estarrón-Espinosa et al. (2009) Tahir, Xiaobo et al. (2017) Pino, Márquez et al. (2006) Tahir, Xiaobo et al. (2017) Tahir, Xiaobo et al. (2017) Chen, Huang et al. (1998)

(Continued)

7.7 Conclusion

Table 7.2 Volatile constituents of Roselle (Hibiscus sabdariffa) calyces. Continued Classes

Compounds

References

4-Methyl-1-(1methylethyl)-3cyclohexen-1-ol acetate Methyl-2-furoate Pentyl propanoate Methyl-2-furoate Methyl salicylate

Chen, Huang et al. (1998)

Methyl hexadecanoate Methyl anthranilate (E)-Phytol acetate Ethyl methyl phenylglycidate

Zannou, Kelebek et al. (2020) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Chen, Huang et al. (1998), Avalos-Martínez, Pino et al. (2019) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Camelo-Méndez, Ragazzo-Sánchez et al. (2013)

Others (E)-Nethole 1-Methyl-4-(1-methylethyl)3-cyclohexenol Caryophyllene Indole γ-Eudesmol γ-Butyrolactone Methacrolein 2,3-Dimethylbutane 1-Propylbenzene Thymoquinone α-Angelica lactone (E)-Anethole Pantolactone Maltol 2-Formylpyrrole

Pino, Márquez et al. (2006) Chen, Huang et al. (1998) Chen, Huang et al. (1998) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Zannou, Kelebek et al. (2020) Tahir, Xiaobo et al. (2017) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Pino, Márquez et al. (2006) Zannou, Kelebek et al. (2020) Zannou, Kelebek et al. (2020) Zannou, Kelebek et al. (2020)

7.7 Conclusion In this chapter, several studies exist which demonstrated the use of Roselle (Hibiscus sabdariffa) as sources of medicinal. The previous studies demonstrated the potential of this plant to be used as a source of health benefits compounds such as organic acid, phenolic acid, flavonoids and anthocyanin for use in food and pharmaceutical industries, among others, further its value as a functional herbal beverage. Taking together, this chapter indicated that aldehydes, ketones, alcohols, terpenes, and esters are the main compounds contributing to the Roselle flavor.

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366. Babalola, S., A. Babalola & O. Aworh (2001). Compositional attributes of the calyces of Roselle (Hibiscus sabdariffa L.). Boll, P. M. (1969). Naturally occurring lactones and lactames. III. The absolute configuration of the hydroxycitric acid lactones: Hibiscus acid and garcinia acid. Acta Chemica Scandinavica, 23, 286
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812. Eggensperger, H., & Wilker, M. (1996). Hibiscus-Extrakt: Ein hautvertra¨glicher Wirkstoffkomplex aus AHA’s und polysacchariden. Teil 1. und Kosmetik, 77(9), 522
523. Escobar-Ortiz, A., Castan˜o-Tostado, E., Rocha-Guzma´n, N. E., Gallegos-Infante, J. A., & Reynoso-Camacho, R. (2020). Anthocyanins extraction from Hibiscus sabdariffa and identification of phenolic compounds associated with their stability. Journal of the Science of Food and Agriculture, n/a(n/a). Farag, M. A., Rasheed, D. M., & Kamal, I. M. (2015). Volatiles and primary metabolites profiling in two Hibiscus sabdariffa (Roselle) cultivars via headspace SPME-GC-MS and chemometrics. Food Research International, 78, 327
335. Fernndez-Arroyo, S., Rodrı´guez-Medina, I. C., Beltra´n-Debo´n, R. l, Pasini, F., Joven, J., Micol, V., . . . Ferna´ndez-Gutie´rrez, A. (2011). Quantification of the polyphenolic

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1495. Gonzalez-Palomares, S., Estarro´n-Espinosa, M., Go´mez-Leyva, J. F., & Andrade-Gonza´lez, I. (2009). Effect of the temperature on the spray drying of Roselle extracts (Hibiscus sabdariffa L.). Plant Foods for Human Nutrition, 64(1), 62
67. Griebel, C. (1939). “Hibiscus” flowers,” a drug used in the preparation of food and drink, its principal component a new acid of fruit acid character (hibiscus acid). Zeit Untersuchung Lebensmittel, 77, 561
571. Herranz-Lpez, M., Ferna´ndez-Arroyo, S., Pe´rez-Sanchez, A., Barrajo´n-Catala´n, E., Beltra´nDebo´n, R. l, Mene´ndez, J. A., . . . Micol, V. (2012). Synergism of plant-derived polyphenols in adipogenesis: perspectives and implications. Phytomedicine: International Journal of Phytotherapy and Phytopharmacology, 19(3-4), 253
261. Hida, H., Yamada, T., & Yamada, Y. (2007). Genome shuffling of Streptomyces sp. U121 for improved production of hydroxycitric acid. Applied Microbiology and Biotechnology, 73(6), 1387
1393. Hossain, M. F., Akhtar, S., & Anwar, M. (2015). Nutritional value and medicinal benefits of pineapple. International Journal of Nutrition and Food Sciences, 4(1), 84
88. Ifie, I., Ifie, B. E., Ibitoye, D. O., Marshall, L. J., & Williamson, G. (2018). Seasonal variation in Hibiscus sabdariffa (Roselle) calyx phytochemical profile, soluble solids and α-glucosidase inhibition. Food Chemistry, 261, 164
168. Indovina, R., & Capotummino, G. (1938). Chemical analysis of karkade, the extract derived from Hibiscus sabdariffa L. cultivated in Sicily (Palermo), and Annali Di Chimica Applicata, 28, 413
418. Ismail, A., Ikram, E. H. K., & Nazri, H. S. M. (2008). Roselle (Hibiscus sabdariffa L.) seeds-nutritional composition, protein quality and health benefits. Food, 2(1), 1
16. Jabeur, I., Pereira, E., Barros, L., Calhelha, R. C., Sokovi´c, M., Oliveira, M. B. P. P., & Ferreira, I. C. F. R. (2017). Hibiscus sabdariffa L. as a source of nutrients, bioactive compounds and colouring agents. Food Research International, 100, 717
723. Jamini, T., Islam, A., Mohi-ud-Din, M., & Saikat, M. (2019). Phytochemical composition of calyx extract of Roselle (Hibiscus sabdariffa L.) genotypes. Journal of Food Technology and Food Chemistry, 2(102), 2. Jirovetz, L., Jaeger, W., Remberg, G., Espinosa-Gonzalez, J., Morales, R., Woidich, A., & Nikiforov, A. (1992). Analysis of the volatiles in the seed oil of Hibiscus sabdariffa (Malvaceae) by means of GC-MS and GC-FTIR. Journal of Agricultural and Food Chemistry, 40(7), 1186
1187. Juhari, N. H., Bredie, W. L. P., Toldam-Andersen, T. B., & Petersen, M. A. (2018). Characterization of Roselle calyx from different geographical origins. Food Research International, 112, 378
389. Juhari, N.H., Varming, C., M.A. Petersen (2015). Analysis of aroma compounds of Roselle by Dynamic Headspace Sampling using different sample preparation methods. XIV Weurman flavour research symposium, Context Products Ltd. Leicestershire, England. Khafaga, E. R., Koch, H., El Afry, M. M. F., & Prinz, D. (1980). Stage of maturity and quality of karkadeh (Hibiscus sabdariffa L. var. sabdariffa). 1. Organic acids. 2. Anthocyanins. 3. Mucilage, pectin and carbohydrates. 4. Improved drying and harvesting systems. Angewandte Botanik, 54(5/6), 287
318. Kouakou, T., Konkon, N., Ayolie´, K., Obouayeba, A., Abeda, Z., & Kone´, M. (2015). Anthocyanin production in calyx and callus of Roselle (Hibiscus sabdariffa L.)

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and its impact on antioxidant activity. Journal of Pharmacognosy and Phytochemistry, 4(3). Kuo, C.-Y., Kao, E.-S., Chan, K.-C., Lee, H.-J., Huang, T.-F., & Wang, C.-J. (2012). Hibiscus sabdariffa L. extracts reduce serum uric acid levels in oxonate-induced rats. Journal of Functional Foods, 4(1), 375
381. Lin-Holderer, J., Li, L., Gruneberg, D., Marti, H. H., & Kunze, R. (2016). Fumaric acid esters promote neuronal survival upon ischemic stress through activation of the Nrf2 but not HIF-1 signaling pathway. Neuropharmacology, 105, 228
240. Luvonga, W.A., M.S. Njoroge, A. Makokha & P.W. Ngunjiri (2010). Chemical characterisation of Hibiscus sabdariffa (Roselle) calyces and evaluation of its functional potential in the food industry. JKUAT Annual Scientific Conference Proceedings. McKay, D. (2009). Can hibiscus tea lower blood pressure. AfroFood Industry Hi-Tech, 20 (6), 40
42. Morton. (1987). Soursop In Fruits of Warm Climates (pp. 75
80). Greensboro, NC: Media Incorporated. Ndiaye, O. (2016). Impacts of Water, Extraction Procedure and Origin on Anthocyanins and Volatile Compositions of Hibiscus Extracts and Freeze-Dried Hibiscus, Virginia Tech. Peng, C.-H., Chyau, C.-C., Chan, K.-C., Chan, T.-H., Wang, C.-J., & Huang, C.-N. (2011). Hibiscus sabdariffa polyphenolic extract inhibits hyperglycemia, hyperlipidemia, and glycation-oxidative stress while improving insulin resistance. Journal of Agricultural and Food Chemistry, 59(18), 9901
9909. Pimentel-Moral, S., Borra´s-Linares, I., Lozano-Sa´nchez, J., Arra´ez-Roma´n, D., Martı´nezFe´rez, A., & Segura-Carretero, A. (2019). Supercritical CO2 extraction of bioactive compounds from Hibiscus sabdariffa. The Journal of Supercritical Fluids, 147, 213
221. Pino, J. A., Ma´rquez, E., & Marbot, R. (2006). Volatile constituents from tea of Roselle (Hibiscus sabdariffa L.), Revista CENIC Ciencias Quı´micas, 37(3), 127
129. Piovesana, A., & Noren˜a, C. P. Z. (2019). Study of acidified aqueous extraction of phenolic compounds from Hibiscus sabdariffa L. calyces. The Open Food Science Journal, 11(1). Ramı´rez-Rodrigues, M. M., Plaza, M. L., Azeredo, A., Balaban, M. O., & Marshall, M. R. (2012). Phytochemical, sensory attributes and aroma stability of dense phase carbon dioxide processed Hibiscus sabdariffa beverage during storage. Food Chemistry, 134 (3), 1425
1431. Ramrezaˆ Rodrigues, M. M., Balaban, M. O., Marshall, M. R., & Rouseff, R. L. (2011). Hot and cold water infusion aroma profiles of Hibiscus sabdariffa: fresh compared with dried. Journal of Food Science, 76(2), C212
C217. Riaz, G., & Chopra, R. (2018). A review on phytochemistry and therapeutic uses of Hibiscus sabdariffa L. Biomedicine & Pharmacotherapy, 102, 575
586. Rodrı´guez-Medina, I. C., Beltra´-Debo´n, R. l, Molina, V. M., Alonso-Villaverde, C., Joven, J., Menendez, J. A., . . . Ferna´ndez-Gutie´rrez, A. (2009). Direct characterization of aqueous extract of Hibiscus sabdariffa using HPLC with diode array detection coupled to ESI and ion trap MS. Journal of Separation Science, 32(20), 3441
3448. Sinela, A., Rawat, N., Mertz, C., Achir, N., Fulcrand, H. l n, & Dornier, M. (2017). Anthocyanins degradation during storage of Hibiscus sabdariffa extract and evolution of its degradation products. Food Chemistry, 214, 234
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Singh, P., Khan, M., & Hailemariam, H. (2017). Nutritional and health importance of Hibiscus sabdariffa: a review and indication for research needs. Journal of Nutritional Health & Food Engineering, 6(5), 00212. Tahir, H. E., Arslan, M., Mahunu, G. K., Mariod, A. A., Wen, Z., Xiaobo, Z., . . . El-Seedi, H. (2020). Authentication of the geographical origin of Roselle (Hibiscus sabdariffa L) using various spectroscopies: NIR, low-field NMR and fluorescence. Food Control, 114, 107231. Tahir, H. E., Xiaobo, Z., Mariod, A. A., Mahunu, G. K., Abdualrahman, M. A. Y., & Tchabo, W. (2017). Assessment of antioxidant properties, instrumental and sensory aroma profile of red and white Karkade/Roselle (Hibiscus sabdariffa L.). Journal of Food Measurement and Characterization, 11(4), 1559
1568. Williamson, E.M., Driver, S., & Baxter, K. (2009). Stockley’s herbal medicines interactions: a guide to the interactions of herbal medicines, dietary supplements and nutraceuticals with conventional medicines/editors, Elizabeth Williamson, Samuel Driver, Karen Baxter; editorial staff, Mildred Davis. . .[et al.], digital products team, Julie McGlashan, Elizabeth King, London; Chicago: Pharmaceutical Press. Zannou, O., Kelebek, H., & Selli, S. (2020). Elucidation of key odorants in Beninese Roselle (Hibiscus sabdariffa L.) infusions prepared by hot and cold brewing. Food Research International, 133, 109133.

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CHAPTER

Oil recovery from Hibiscus sabdariffa seeds

8

Abdalbasit Adam Mariod1,2, Haroon Elrasheid Tahir3 and Gustav Komla Mahunu4 1

Indigenous Knowledge and Heritage Center, Ghibaish College of Science & Technology, Ghibaish, Sudan 2 College of Sciences and Arts-Alkamil, University of Jeddah, Alkamil, Saudi Arabia 3 School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China 4 Department of Food Science and Technology, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana

Chapter Outline 8.1 Introduction .................................................................................................113 8.2 Roselle components as food source ...............................................................114 8.3 Protein of Roselle seeds ...............................................................................115 8.4 Oil of Roselle seeds ......................................................................................115 8.5 Roselle oil seed products ..............................................................................120 8.6 Conclusion ...................................................................................................121 References ..........................................................................................................121

8.1 Introduction Hibiscus sabdariffa (Roselle) is a well-known worldwide crop. There are more than 300 species which are distributed across the world in tropical and subtropical regions. In a warmer and more humid climate Roselle can adapt to a variety of soils (Singh, Khan, & Hailemariam, 2017). Roselle is widely grown in different environments, and is an underused multipurpose crop which provides food and cash income for farmers. Roselle is renowned for its edible fleshy calyces and leaves used to make salads, tea, juices, jams and many other items (Islam, Jamini, Islam, & Yeasmin, 2016). Almost all parts of the hibiscus plant, from the leaves to the roots, have a wide range of uses in various foods and in alternative medicine. Hibiscus extracts play a prominent role in treating many diseases such as heart disease and cancer, and hibiscus has antioxidant activity and is used in the elimination of obesity (Singh et al., 2017). The plant has beautiful red flowers and is grown for decoration or for sepal Roselle (Hibiscus sabdariffa). DOI: https://doi.org/10.1016/B978-0-12-822100-6.00001-X © 2021 Elsevier Inc. All rights reserved.

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production that surround the flower and after drying it is either dark red or light red. Boil these dry, red sepals and drink their sour-flavored infusion hot after adding sugar to it or after cooling and sweetening it as a refreshment rich in vitamin C. Hibiscus infusion without boiling lowers high blood pressure, increases the speed of blood circulation, helps in strengthening the heartbeat and kills microbes, which makes it useful in treating fevers and microbial infections as it is acidic, tonic and aid for digestion, but it is cautioned against using it for those with low blood pressure. It is used as a coloring pigment in the chemical and cosmetic industries and in the food industries as a natural and healthy colorant, and it is also a general tonic (https://www.biodiversitylibrary.org).

8.2 Roselle components as food source Hibiscus provides the body with many nutrients, such as, calcium, iron, and vitamin B2, and moderate amounts of antioxidants, such as, vitamin C, and anthocyanin, which provides it with red or purple color, and helps its antioxidant content for protection against damage resulting from oxidation of free radicals, which is one of the most important reasons for raising the risk of many types of cancer, such as, stomach cancer and blood cancer, and it is believed that antioxidants may reduce the risk of developing cardiovascular disease by reducing oxidation of harmful cholesterol in the body (DMello, 2020). Hibiscus seeds and their extracts offer many benefits to the human body, and some of them are as follows: •

•

A rich source of protein: Hibiscus seeds are one of the sources rich in protein, and some amino acids such as lysine, arginine, and leucine, phenylalanine and glutamic acid. Antioxidant effect: The extracts of the seeds of the hibiscus plant have an antioxidant effect, and this effect increases when the antioxidants in the extracts are bound to other antioxidants (Shaheen, El-Nakhlawy, & Al-Shareef, 2012).

In Africa, hibiscus seeds are used as an additive to meals after grinding them or as an alternative to coffee. The dried red calyces from the plant are used to prepare tea or acidic juice after adding sugar, and these calyces are also used as a colorant in food. The oil extracted from hibiscus seeds is used in cooking. Although there are studies indicated that Roselle oil contains some toxic substances, it is advisable to use it in the manufacture of soap in the field of cosmetics. Hibiscus is a short daytime plant and is cultivated early during June in tropical regions. Hibiscus calyces contain pectin that can be used in making jams and jelly, besides they are good sources of calcium, iron, vitamins such as niacin and riboflavin. Roselle calyces contain antioxidants, anthocyanins which inhibit lipid peroxidation. Fresh leaves contain 2%
3% protein and contain traces of calcium, phosphorus, and iron (https://www.echocommunity.org). There are many organic acids

8.4 Oil of Roselle seeds

in Roselle drink, including citric, malic, and tartaric acid. Hibiscus contains various compounds of medical importance known as photochemicals with nutritional and medicinal value (Singh et al., 2017).

8.3 Protein of Roselle seeds Protein is the most important element in the body, and it is present in every cell and one of the main components in bones, muscles, cartilage, skin and blood, as approximately 20% of the body consists of protein, so it is necessary to maintain the daily intake of protein to stay in good health, as well. Protein intake is crucial when trying to lose weight and build muscle. Examples of plant protein are peas, spinach, lentils, corn, chickpeas, etc. (Gallagher et al., 2000). Hibiscus is grown in some African countries and the Indian peninsula, the seeds are a good source of protein, and instead of using traditional sources of protein, protein concentrates and isolates are useful for their low cost when used as food additive, which contributes to addressing the problem of protein deficiency (Deshpande et al., 2000). Halimatul, Amin, Mohd.-Esa, Nawalyah, and Siti Muskinah (2007) studied the Malaysian hibiscus seeds and found that the protein is similar to casein. In addition, they confirmed that the quality of the hibiscus protein increases after boiling. This research group suggested that the hibiscus seed protein could be a suitable alternative to the dietary protein. Mabrouk, Amin, Youssef, and Abou- Samaha (2018) reported 39.47% of essential amino acids of Roselle seeds and that, methionine and cystine were the limiting amino acids. These authors fractionated Roselle seed protein, utilizing sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, and they reported 10 bands ranged in molecular weight from 20 to 245 kDa.

8.4 Oil of Roselle seeds The oil content of karkade calices ranged from 0.9% to 1.15%, depending on the season, the oil content of karkade seeds ranged from 19.46% to 24.04%, the oil content of karkade leaves ranged from 5.35% to 7.31% while the oil content of karkade roots ranged from 0.65%
0.80% and the oil content of karkade stems ranged from 3.95% to 4.32% (AL Shoosh, 1997). Islam et al. (2016), reported fat content of 2.61% for fresh calyces, 0.30% for fresh leaves, and almost more than 16.0% for seed. The physical and chemical properties of Roselle seed (RS) oil suggests that it could have several important industrial applications as an ingredient in cooking, cosmetics and paint industries and justify its added value for cultivation (Mohamed, Fernandez, Pineda, & Aguilar, 2007). Seed oil can also be used to produce biodiesel (Nakpong & Wootthikanokkhan, 2010). Roselle seed oil is low

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in cholesterol and rich in tocopherols, β-sitosterol and γ-tocopherol. Mohamed et al. (2007), analyzed phytosterols of Roselle seed oil and reported 71.9% as β-sitosterol, 13.6% as campesterol, 5.9% as Δ-5-avenasterol and 1.35% as cholesterol. Total tocopherols were detected at an average concentration of 2000 mg/kg, including α-tocopherol (25%), γ-tocopherol (74.5%), and δ-tocopherol (0.5%). El-Deab and Ghamry (2017) carried a qualitative estimation of tocopherols in Roselle seed oil, their results showed the presence of α, δ and γ -tocopherol. γ -tocopherol was the highest, and almost twice as much as α-tocopherol, while δ-tocopherol was barely detected. Tocopherols are naturally occurring constituents found in vegetable oils in varying amounts. The presence of these compounds is important in relation to oil stability and nutritional labeling. Mahmoud, Selim, and Abdel-Baki (2008) reported that Roselle seeds contained relatively high fat and protein contents (20.97% and 29.61%, respectively). While Nasrabadi, Zarringhalami, and Ganjloo (2018) reported that the Roselle seeds are the main sources of protein (26.6%), lipid (21.0%) and fiber (19.8%), respectively. These variations in the composition content of Roselle seed might be due to varieties and ecological conditions. The oil content of Roselle seed was reported to be 20%, related to aqueous extract, however, the total phenolic content was 201 mg gallic acid equivalents (GAEs)/100 g, the extracted compounds using acetone showed antioxidant activity of 94.2% measured using 2,2-diphenyl1-picrylhydrazyl, and 75.9% using 2, 20 -azinobis (3-ethylbenzothiazoline-6-sulfonic acid) techniques. The physico-chemical parameters of crude cold extracted oil with yellow-greenish color showed 1.4674 as refractive index, 0.78% as saponification value, 196.82 as iodine value, 97.62 (g of I2/100 g oil) as unsaponifiable matter, with low peroxide value of 1.52%. Gas liquid chromatography (GLC) technique is always used for identification and quantitative determination of fatty acids. Nasrabadi et al. (2018) used GLC and reported that, the major fatty acids found in Roselle seed oil were oleic acid (38.46%), linoleic (33.25%), palmetic (20.52%) and stearic acids (5.79%), that was in agreement with that reported by AL Shoosh (1997) who reported that, the main fatty acid composition of Roselle seed oil included myristic acid (trace), palmitic acid (26.38%), stearic acid (4.80%), oleic acid (35.84%), and linoleic acid (32.97%) but it was in contrast with Nasrabadi et al. (2018) who reported that the main fatty acids in the Roselle seeds oil were linoleic (41.06% 6 0.7%), oleic (27.07% 6 0.01%) and palmitic (21.9% 6 0.03%) acids. In relation to the fatty acid composition Al Shoosh (1997) reported that the stability of the karkade seed oil as compared to other edible Sudanese oils (peanut, sesame, cottonseed and sunflower) was fairly good. Mabrouk et al. (2018) reported that, the crude RS oil consists of eight lipid classes, which are glyceride and nonglyceride compounds polar lipids, monoacylglycerols, 1,2 and 2,3 diacylglycerols, 1,3 diacylglycerols, free fatty acids, triacylglycerols, hydrocarbons, and sterol esters. Gas chromatography (GC) analysis of crude RS oil indicated that it composed of 30.3% saturated fatty acids, (palmitic and stearic) and 69.7% of unsaturated fatty acids, in which oleic acid was 31.27%, while linoleic acid was 37.61%. Physicochemical analysis of crude RS

8.4 Oil of Roselle seeds

oil showed that it had a yellow color. Mineral elements of edible oils perform important and vital functions for the body that can be summarized as follows: formation of bones and teeth, due to the presence of calcium, magnesium, phosphorus. It is involved in the formation of soft tissues. It enters into the formation of hormones, enzymes, and vitamins, and works to activate them. Calcium and potassium help in regulating the transmission of nerve impulses in the central nervous system, as well as the regulation of heart rhythms. Sodium and potassium regulate the water balance inside and outside the cells of the body to prevent dehydration (Harry, Preuss, & Macn, 2020). Crude RS contains the following minerals: K, Mg, P, Ca, Fe, Cu, Zn. Potassium (1140 mg/100 g) was the predominant element in the seeds, followed by phosphorus (482 mg/100 g), magnesium (242 mg/100 g), and calcium (239 mg/100 g). One of the chemical properties of Roselle oil is that it is a stable oil and its ground seeds are not affected much by the enzymatic analysis. The oil is characterized by a fairly high resistance to oxidation and spoilage. The color of the crude oil can be affected if it is exposed to a temperature of 170  C, so care must be taken not to raise temperature more than 150 C during oil processing steps, for example, the neutralization of free acids, shortening the color and removing the odor (deodorization). Hibiscus seed oil is used after being purified as an edible oil, and it has been used in frying foods. It is an oil with a normal taste, and its color is similar to refined cottonseed oil. Roselle seed oil is of unsaturated type and can be classified in the oleic
linoleic acid group (Nzikou et al., 2011). Al-Okbi, Abdel-Razek, Mohammed, and Ottai (2017) studied an Egyptian Roselle seed oil extracted by cold press, they found that the principal fatty acids were linoleic (37.11%), oleic (33.08%), palmitic (17.19%), and stearic (7.96%). The percentage of saturated fatty acids, monounsaturated fatty acids, and polyunsaturated fatty acids were 27.49%, 33.93%, and 38.58%, respectively, with total unsaturated fatty acids as 72.51%. These authors reported lauric, myrstic, palmitoleic, linolenic, arachidic and gondoic as the minor fatty acids (Al-Okbi et al., 2017). Oxidation stability is one of the most important quality parameters of edible vegetable oils. It determines their usefulness in technological processes as well as shelf life. Rancimat test, Schaal Oven Test—thermostatic test and chemical determinations of peroxide value, anisidine value, and acid value are the main methods used to determine the oxidative stability of oils. The Rancimat test determines the oxidation stability of oils in a very short time, however, high temperatures and intensive aeration are used, which change the nature of the oxidation process (Magdalena et al., 2018). Stability of crude Roselle seed oil against oxidation during the accelerated storage of oil indicated that the crude oil had induction period 10 days at 65 C. The relatively high fat content of seeds, high protein content of resulted meal for animal feeding and/or possible human use beside, the relatively high oxidation stability of tested oil suggest that the Roselle seeds could be novel and economic

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source of healthy edible fat and other food industry applications (Mahmoud et al., 2008). The cold pressed Roselle seed oil showed to possess high safety and in vitro antioxidant activity. The cold pressed Roselle seed oil contains alpha, beta, gamma, and delta tocopherols with the highest content of gamma tocopherol (67.58 mg 6 0.16/100 g oil). The oil also contained alpha, beta and gamma tocotrienols where alpha tocotrienol was the major (2.11 mg 6 0.03/100 g oil). Total tocopherol content was 96.2 mg 6 0.26/100 g oil while total tocotrienols concentration was 3.48 mg 6 0.03/100 g oil. The presence of tocopherols and tocotrienols in cold pressed oil Roselle seed oil renders the oil both high oxidative stability and health benefits as antioxidant. Gamma-tocopherol had the most considerable fraction (70% of total tocopherols), almost more than twice of alpha-tocopherol (28.6%), while beta-tocopherol was hardly detected. Alphatocotrienols represent the abundant fraction (61%) of the total tocotrienols followed by gamma and beta (24.4% and 14.9%, respectively). Total phenolic and flavonoids contents were 56.31 mg GAE and 4.99 mg catechin gG1 oil, respectively. Induction period of oxidative stability of Roselle seed oil was 24.88 hours. (Al-Okbi et al., 2017). Degradation of edible oil toward rancidity is wasteful and/or health challenging. In this study, oil extracted from Roselle seeds was analyzed for elemental and microbial content. Roselle oil under different conditions of storage were progressively studied for 16 weeks to investigate the changes in the physicochemical properties signaling deterioration. Fourier transformed infrared (FTIR) analysis before and after the storage period were carried out to investigate the changes in the functionality of the oils. The changes in the FTIR spectra over the storage period revealed different rates and fashions of deterioration. From the study, the conditional degradation of Roselle seed oil was least severe in the dark air-tight/ covered storage conditions (Nkafamiya, Atiku, Akinterinwa, & Fari, 2017). The Roselle seeds as by-product could be economic and novel source of edible fat, helping for covering a part of the deficiency in edible oils. In addition to the characteristics of linoleic
oleic fatty acid and relatively high stability against oxidation makes Roselle seed oil very useful as for healthy edible oil and some food industry application (Mahmoud et al., 2008). Naeem, Zahran, and Hassanein (2019) carried out a distinguished study on Roselle seed oil using three green extraction methods including supercritical CO2 (SC-CO2), screw, and hydraulic press, and compared them to conventional one. This research group studied the amount of oil, phenolic compounds, oxidative activity, fatty acids and tocopherols, and the relationship of these components with the stability of hibiscus seed oil, their results showed that the oil extracted with solvent gave a high percentage of oil and low acidity and peroxide value, oil extraction with carbon dioxide gave a high percentage of fatty acids, followed by cold press and solvent extraction. The highest degree of stability and the highest amount of phenolic compounds and antioxidants are found in the oil extracted by means of carbon dioxide, followed by the oil extracted by hydraulic squeezing, then the oil extracted by the solvent and finally the oil extracted by the hydraulic

8.4 Oil of Roselle seeds

press. Cold pressing gave a higher percentage of oil than extraction with carbon dioxide, as it is more economical than other methods. Mohamed et al. (2007) studied the antioxidant activity for different parts of the hibiscus plant, including the seeds, stems, leaves and sepals, taking into account their oxidative energy dissolved in water, which is soluble in lipids, and the content of tocopherols. These researchers extracted hibiscus oil and described its properties and explained that the acidity number is 2.24% and the peroxide value is 8.6 mg equivalent per kilogram, and its stability was 15 hours, the refractive index 1.477, the density of 0.92 kg/L, Oleic 28%, palmitic 20%. Mohamed et al. (2007) suggested that Roselle (Hibiscus sabdariffa) could be used as a source of strong water-lipid soluble antioxidants, including vitamin E. These seeds are a source of a vegetable oil that is low in cholesterol and rich in other phytosterols and tocopherols, particularly β-sistosterol and γ-tocopherol. Fiad (1991) used thin-layer chromatography to fractionate the hibiscus phospholipids, and then separated them using salicylic acid and showed that they were cephalin, lithicin, and phosphatidic acid. This research group prepared phospholipid acids by means of transesterification and found different proportions of palmitic, oleic and linoleic acid. Rukmini, Vijayaraghavan, and Tulpule (1982) reported that, Hibiscus sabdariffa seed oil, contains cyclopropenoid fatty acids (2.9%) and epoxy fatty acids (2.6%) in addition to normal fatty acids found in vegetable oils. There are several nutritional and toxicological studies conducted on hibiscus oil by adding 10% to a rat meal that contains 20% protein with an adequate amount of minerals and vitamins. This study revealed that mice fed hibiscus oil had weak growth and reproductive appearance, and also showed a marked change in the metabolism of the liver, which confirms that crude or refined hibiscus oil is not suitable for human consumption. Atta and Imaizumi (2002) studied the Roselle seed oil properties they reported melting point as
1.1 6 0.3 C, 1.0% unsaponifable matter and 109 as iodine. So they consider it as semi-dry oil, they reported also 69.2% triacyglycerols and sterols 3.5%. GLC analysis of Roselle seed oil showed it contain 45.3% linoleic, 27.2% oleic and 17.3% palmitic acids as the predominant fatty acids. Roselle seed oil is thought to be similar to corn oil in chemical composition and lipid fractions, Roselle seed oil might provide a new source of edible oil (Atta & Imaizumi, 2002). The percentage value of triacylglycerol, free fatty acids and phospholipid fractions in Roselle seed oil were 69.2%, 3.5%, and 1.9%, respectively, besides 1,3 diacylglycerol. The data showed that triacylglycerol was the main lipid fraction of Roselle seed oil. Crude Roselle seed oil contained 559 mg β-sitosterol per 100 g oil. These amounts were bigger than those detected in peanut and unrefined olive oils. β-sitosterol has an inhibitory effect on colon cancer. Campesterol was 107, stigmasterol was 50.6 and β-sitostanol was 39.9 mg/100 g in Roselle seed oil (Atta & Imaizumi, 2002).

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Hussein et al. (2020) investigated the possible mechanism of hepatoprotective potential of Roselle seed oil (RSO) and its nano-formulation (RSO-NE) in a paracetamol-induced hepatotoxicity rat model, compared to silymarin. These authors found that, Roselle seed oil and its nano-formulation protected the liver against paracetamol-intoxication and maintained the overall architecture of liver tissues in a dose dependent manner. These authors concluded that linoleic, oleic, palmitic and stearic acids, n-eicosane, β-sitosterol and tocopherols were the major constituents, which contributed synergistically to its protective effect. These findings encourage the use of RSO in development of health promoting products such as food supplements, functional food and nutraceuticals for the prevention of liver disease (Hussein et al., 2020).

8.5 Roselle oil seed products Use of Roselle seed oil for production of mayonnaise for improving its oxidative stability was investigated by El-Deab and Ghamry (2017). Their data demonstrated that mayonnaise containing 50% Roselle seed oil was superior in sensory characteristics and oxidative stability as compared with control. Biodiesel is very important to meet the future energy shortage. Sahu, Tiwari, and Mishra (2017) found that physicochemical properties of biodiesel produced from Roselle are very close to fossil diesel. These authors used Roselle oil biodiesel alone and blended with petroleum diesel, they confirmed that the produced products were approved and meet ASTM standards for safe operation in any compression-ignition engine designed to be operated on petroleum diesel. Roselle seeds-based biodiesels seems to be a bit superior to other biodiesels. With about 30%
35% of “oil value,” Roselle oil shows good conversion rate to bio diesels. A liter of Roselle seed based vegetable oil converts to approximately 700 ml of bio fuels apart from glycerin. And the most satisfactory part is that when mixed 5% of methanol or ethanol with diesel, the performance of a diesel engine could be increased, especially in the high load conditions. Thermal efficiency among all blends increased when load is increased but pure diesel shows better Brake Specific Fuel Consumption than other blends (Sahu et al., 2017). Hibiscus seeds contain a variety of bioactive phytochemicals like polyphenols, phenolic acids, phytosterols, amino acids, fatty acids, phospholipids, etc., which possess nutritional and phytotherapeutic values. Hibiscus seeds are also a good source of dietary fiber and protein which has great promise as a fodder source for livestock and value-added nutritional foods like defatted seed meal, protein concentrates from defatted seeds, seed cake, etc. The concentration of more oil content was changed towards the development of low cyclopropenoid fatty acids, epoxy fatty acids, and palmitic acid content (saturated) along with increased polyunsaturated omega acids as content varieties (Dhar, Kar, Ojha, Pandey, & Mitra, 2015).

References

8.6 Conclusion Hibiscus oil is considered one of the future oils due to the growing areas of cultivation of the hibiscus crop and the great demand to benefit from the waste of the sepals and petals processing in the food industries, as the seeds are considered a by-product that cannot be used. Roselle seed oil can be used in the manufacture of cooking oil, mayonnaise, and biodiesel.

References AL Shoosh, W. G. A. (1997). Chemical composition of some Roselle (Hibiscus sabdariffa) genotypes (M.S. thesis). Khartoum, Sudan: Faculty of Agriculture University of Khartoum. Al-Okbi, S. Y., Abdel-Razek, A., Mohammed, S. E., & Ottai, M. E. (2017). Roselle seed as a potential new source of healthy edible oil. The Journal of Biological Sciences, 17, 267
277. Atta, M. B., & Imaizumi, K. (2002). Some characteristics of crude oil extracted from Roselle (Hibiscus sabdariffa L.) seeds cultivated in Egypt. Journal of Oleo Science, 51 (7), 457
461. Deshpande, S. S., Salunkhe, D. K., Oyewole, O. B., Azam-Ali, S., Battock, M., & Bressani, R. (2000). Fermented grain legumes, seeds and nuts: A global perspective. FAO agricultural services bulletin (142, pp. 1
53). Rome, Italy: FAO. Dhar, P., Kar, C. S., Ojha, D., Pandey, S. K., & Mitra, J. (2015). Chemistry, phytotechnology, pharmacology and nutraceutical functions of kenaf (Hibiscus cannabinus L.) and Roselle (Hibiscus sabdariffa L.) seed oil: An overview. Industrial Crops and Products, 77, 323
332. DMello, S. (2020). Health benefits of hibiscus. Retrieved from www.medindia.net. (Accessed 12 September 2020). El-Deab, S. M., & Ghamry, H. E. (2017). Nutritional evaluation of Roselle seeds oil and production of mayonnaise. International Journal of Food Science and Nutrition Engineering, 7(2), 32
37. Available from https://doi.org/10.5923/j.food.20170702.02. Fiad, S. (1991). Phospholipids of six seed oils of malvaceae. The Journal of the American Oil Chemists’ Society, 68(1), 26
28. Gallagher, D., Heymsfield, S. B., Heo, M., Jebb, S. A., Murgatroyd, P. R., & Sakamoto, Y. (2000). Healthy percentage body fat ranges: An approach for developing guidelines based on body mass index. The American Journal of Clinical Nutrition, 72(3), 694
701. Halimatul, S. M. N., Amin, I., Mohd.-Esa, N., Nawalyah, A. G., & Siti Muskinah, M. (2007). Protein quality of Roselle (Hibiscus sabdariffa L.) seeds 131 short communication protein quality of Roselle (Hibiscus sabdariffa L.) seeds. ASEAN Food Journal, 14 (2), 131
140. Harry, G., Preuss, M. D., & Macn, C. N. S. (2020). Sodium, chloride, and potassium. In B. P. Marriott, D. F. Birt, & A. A. Yates (Eds.), Present knowledge in nutrition (Eleventh Edition) Volume 1: Basic nutrition and metabolism (pp. 467
484). USA: International Life Sciences Institute (ILSI). Published by Elsevier Inc.

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Hussein, M. E., El Senousy, A. S., Abd-Elsalam, W. H., Ahmed, K. A., El-Askary, H. I., Mouneir, S. M., & El Fishawy, A. M. (2020). Roselle seed oil and its nano-formulation alleviated oxidative stress, activated Nrf2 and down regulated m-RNA expression genes of pro-inflammatory cytokines in paracetamol-intoxicated rat model. Records of Natural Products, 14(1), 1
17. Islam, A. K. M. A., Jamini, T. S., Islam, A. K. M. M., & Yeasmin, S. (2016). Roselle: A functional food with high nutritional and medicinal values. Fundamental and Applied Agriculture, 1(2), 44
49. Mabrouk, O. M., Amin, W. A., Youssef, A. M. M., & Abou-Samaha, O. R. (2018). Roselle (Hibiscus sabdariffa) seeds and kernels as a potential source of oil, protein and minerals. Egyptian Journal of Food Science, 46, 55
67. Magdalena, M., Florowska, A., Dłuzewska, E., Wroniak, M., Marciniak-Lukasiak, K., & Zbikowska, A. (2018). Oxidative stability of selected edible oils. Molecules, 23, 1746. Available from https://doi.org/10.3390/molecules23071746. Mahmoud, A. A., Selim, K. A., & Abdel-Baki, M. R. (2008). Physico-chemical and oxidative stability characteristics of Roselle (Hibiscus sabdariffa L.) seed oil as by-product Egypt. Journal of Applied Sciences, 23(7), 247
257. Mohamed, R., Fernandez, J., Pineda, M., & Aguilar, M. (2007). Roselle (Hibiscus sabdariffa) seed oil is a rich source of gamma-tocopherol. Journal of Food Science, 72(3), 207
211. Naeem, M. A., Zahran, H. A., & Hassanein, M. M. M. (2019). Evaluation of green extraction methods on the chemical and nutritional aspects of Roselle seed (Hibiscus sabdariffa L.) oil. OCL - Oilseeds & Fats, Crops and Lipids, 26(33), 1
9. Nakpong, P., & Wootthikanokkhan, S. (2010). Roselle (Hibiscus sabdariffa L.) oil as an alternative feedstock for biodiesel production in Thailand. Fuel, 89, 1806
1811. Nasrabadi, Z. M., Zarringhalami, S., & Ganjloo, A. (2018). Evaluation of chemical, nutritional and antioxidant characteristics of Roselle (Hibiscus sabdariffa L.) seed. Nutrition and Food Sciences Research, 5(1), 41
46. Nkafamiya, I. I., Atiku, J., Akinterinwa, A., & Fari, A. (2017). Effect of storage conditions on the degradation of Roselle (Hibiscus sabdariffa) seeds oil. The International Journal of Biological and Chemical Sciences, 11(3), 1350
1360. Nzikou, J. M., Bouanga-Kalou, G., Matos, L., Ganongo-po, F. B., Mboungou-Mboussi, P. S., Moutoula, F. E., . . . Desobry, S. (2011). Characteristics and nutritional evaluation of seed oil from Roselle (Hibiscus sabdariffa L.) in Congo-Brazzaville. Current Research Journal of Biological Sciences, 3, 141
146. Rukmini, C., Vijayaraghavan, M., & Tulpule, P. G. (1982). Nutritional and toxicological evaluation of Hibiscus sabdariffa oil and Cleome viscosa oil. Journal of the American Oil Chemists’ Society, 59(10), 415
419. Sahu, A., Tiwari, J. K., & Mishra, S. (2017). Performance and experimental analysis of Roselle oil as bio-diesel blend on four stroke, diesel engine. International Journal of Innovative Research in Science, Engineering and Technology, 6(9), 18072
18080. Available from https://doi.org/10.15680/IJIRSET.2017.0609045. Shaheen, M. A., El-Nakhlawy, F. S., & Al-Shareef, A. R. (2012). Roselle (Hibiscus sabdariffa L.) seeds as unconventional nutritional source. African Journal of Biotechnology, 11(41), 9821
9824. Singh, P., Khan, M., & Hailemariam, H. (2017). Nutritional and health importance of Hibiscus sabdariffa: A review and indication for research needs. Journal of Nutritional Health and Food Engineering, 6(5), 125
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Food use of whole and extracts of Hibiscus sabdariffa

9

Gustav Komla Mahunu1, Haroon Elrasheid Tahir2, Mildred Osei-Kwarteng3, Abdalbasit Adam Mariod4,5 and Joseph Patrick Gweyi-Onyango6 1

Department of Food Science and Technology, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana 2 School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China 3 Department of Horticulture, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana 4 Indigenous Knowledge and Heritage Center, Ghibaish College of Science & Technology, Ghibaish, Sudan 5 College of Sciences and Arts-Alkamil, University of Jeddah, Alkamil, Saudi Arabia 6 Department of Agricultural Science and Technology, Kenyatta University, Nairobi, Kenya

Chapter Outline Abbreviations ......................................................................................................123 9.1 Introduction .................................................................................................124 9.2 Use of whole Roselle plant constituents in food .............................................124 9.2.1 Calyx ..........................................................................................124 9.2.2 Leaves .......................................................................................129 9.2.3 Roselle seeds ..............................................................................129 9.3 Other functional food applications Roselle extracts ........................................130 9.4 Conclusion ...................................................................................................131 References ..........................................................................................................132

Abbreviations FAO UN

Food and Agriculture Organization United Nations

Roselle (Hibiscus sabdariffa). DOI: https://doi.org/10.1016/B978-0-12-822100-6.00009-4 © 2021 Elsevier Inc. All rights reserved.

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9.1 Introduction Over the years, interest in food diversity for improved health and nutrition has increased in favor of plant-based natural food products with high bioactive or aromatic compounds. These compounds have found their source in various plant extracts. Fruits, vegetables, and herbs are the primary sources of these extract compounds. For several years now, most of these plants and their extracts have found their usefulness as food flavors, preservatives, and traditional medicines or in cosmetic industries (Gyawali & Ibrahim, 2014; Tajkarimi, Ibrahim, & Cliver, 2010). Roselle (Hibiscus sabdariffa) is in the list of valuable plants that has gained immediate attention in the face of several reported health benefits to consumers (Juhari, Bredie, Toldam-Andersen, & Petersen, 2018; Tuan Azlan, Hamzah, & Abd Majid, 2020). The rich vitamins and polyphenol in Roselle are capable of stimulating antioxidant properties (Jabeur et al., 2017; Riaz & Chopra, 2018). With respect to the reports of the Food and Agriculture Organization (FAO) of the United Nations (UN), Roselle in recent years has found its way into the international market with a corresponding demand of about 5000 metric tons/year for its various products. Several countries mainly China, Thailand, Sudan, Mexico, Egypt, Senegal, Tanzania, Mali, and Jamaica have benefited from the production and trading of Roselle (Mohamad, Nazir, Rahman, & Herman, 2002; Plotto, Mazaud, Ro¨ttger, & Steffel, 2004). The different parts of the Roselle plant, including the seeds, leaves, fruits, and roots, have diverse uses in food, medicine, and product manufacturing. Therefore this chapter review explored all current and relevant applications of the whole plant and its extracts in food.

9.2 Use of whole Roselle plant constituents in food Generally, the value addition to cultivated and native plant products with medicinal and nutritional properties is rising in response to consumer demand. Through various improved extraction and purification processes, the bioactive compounds such as organic acids, phytosterols, and polyphenols (including those with antioxidant properties) have been identified in plants with diverse uses including phytochemicals in producing dietary supplements or nutraceuticals, functional food constituents, food additives, and medicinal and cosmetic products.

9.2.1 Calyx Whole fruits also considered as of the Roselle plant are mostly picked at the stage when they are tender, plump, fleshy, crisp, and deep red. The chemical structure of some main constituents of the Roselle flowers. From the calyces, various food products such as jam, jellies, sauces, pickles, juices, sirup, teas, and other bakery products are obtained (Da-Costa-Rocha, Bonnlaender, Sievers, Pischel, & Heinrich, 2014).

9.2 Use of whole Roselle plant constituents in food

Hot and cold beverages or drinks are produced from the mature and ripe calyces. The juice mixed with salt, pepper, and molasses can control coughs and biliousness and also help the body to increase stamina and balance electrolyte after sports (Mohamed, Sulaiman, & Dahab, 2012). The beverages are among the popular health drink products recognized by consumers because of their high vitamin C and anthocyanin contents (Eslaminejad & Zakaria, 2011). Medically, the calyx functions as an antispasmodic, hypotensive, and antimicrobial agent and for easing of the uterine muscle (Khalid, Abdalla, Abdelgadir, Opatz, & Efferth, 2012). Also, the calyx is a major source of flavonoids (anthocyanins, anthocyanidins, quercetin), phenolic compounds (protocatechuic acid, eugenol), organic acids and their derivatives (malic, citric, oxalic, tartaric, hibiscus acid), vitamins (ascorbic acid, thiamine, riboflavin), and β-carotene (Da-Costa-Rocha et al., 2014). The anthocyanins and proanthocyanidins are bioactive compounds in the Roselle calyces with the function of reducing blood pressure in humans (Alarco´n-Alonso et al., 2012; Morton, Roselle, Morton, & Dowling, 1987). Researchers have established that the quercetin present in the Roselle plant influences the vascular endothelium. The presence of oxide nitric in Roselle plays the role of increasing renal vasodilatation and kidney filtration (Alarco´n-Alonso et al., 2012). Consistently, reports have shown that extract of Roselle calyces comprise inherent natural and pharmacological properties; where they act as antihypertensive, antidiabetic, antiobesity, antioxidant, hepatoprotective, nephroprotective, diuretic, anticholesterol, and antianaemic (Fig. 9.1) (Laikangbam & Devi, 2012;

FIGURE 9.1 Summary of various bio-functional and pharmacological characteristics of Roselle.

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Mossalam, Abd-El Aty, Morgan, Youssaf, & Mackawy, 2011). The Roselle calyx extract is considered for promoting health benefit as an anticancer agent and capable of reducing chronic diseases such as hypertension, diabetes mellitus, and cardiovascular disease (Formagio et al., 2015; Mardiah, Prangdimurti, & TIP, 2015). It was also reported that phenolic compounds (including flavonoids and cyaniding) perform antimicrobial function, where for instance flavonoid with its phenolic chain may decrease the iron level and increases hydrogen level and, finally, neutralizes bacterial enzymes (Alshami & Alharbi, 2014). Moreover, Sulistyani, Fujita, Miyakawa, and Nakazawa (2016) reported that flavonoids are also thought to have the ability to inhibit the formation of bacterial biofilms. This is possible because the phenolic group in the extract can bind strongly to proteins and enzymes from the bacteria. This activity stops the bacteria from producing biofilm (Sulistyani et al., 2016). This outcome is essential, considering the fact that Staphylococcus epidermidis and some gram-positive and gram-negative bacteria are capable of producing biofilms. So, Roselle calyx extract can be applied as an alternative treatment for infections caused by S. epidermidis (Lusida, Hermanto, & Sudarno, 2017). The juice from the Roselle was found to have the potential of an excellent transporter in the development of functional beverages with gibberellic acid acting as a prebiotic source. Food products from the Roselle are classified as functional foods, thus provide consumers with the required health benefits by counting on the significant contribution of their phytochemicals (Cid-Ortega & Guerrero-Beltra´n, 2015). More so, the safety and functional benefits of natural colorants against the synthetic types brought into focus the search for sources of novel natural colorants and the authentication of the safety parameters of those products already available. This may partly explain the substantial economic potential placed on most tropical plants such as the Roselle. Color is an important quality attribute that determines consumer acceptability of food. It gives the consumer the first impression of the quality of food. Without the addition of colorants, many convenient foods (confectionery products, gelatin desserts, snacks, cake, pudding, ice cream, and beverages) would be colorless and virtually unattractive (Hirunpanich et al., 2006), delaying product sales. The anthocyanins in Roselle calyces have been recommended as water-soluble natural food colorants to replace red artificial coloring additives for the food industry (Jabeur et al., 2017). The synthetic red color serves as emulsifier for carbonated drinks, jam manufacture, juices, and natural food colorants (Duangmal, Saicheua, & Sueeprasan, 2008). The presence of rich anthocyanin, as well as ascorbic acid and hibiscus acid in the calyces, makes it taste sour and agreeable acidic taste and also helps in digestion (Shruthi et al., 2016). The Roselle calyx was found to contain the compounds that account for the presence of red color in the plant; these compounds include delphinidin-3-glucoside, cyaniding-3-glucoside, delphinidin-3-sambubioside, and cyaniding-3-sambubioside (Borra´s-Linares et al., 2015). According to Amor and Allaf (2009), delphinidin-3-sambubioside (Dp-3-sam) (70% of the anthocyanins) and cyanidin-3-sambubioside (Cyn-3-sam) are the significant pigments, whereas

9.2 Use of whole Roselle plant constituents in food

delphinidin-3-glucoside (Dp-3-glu) and cyanidin-3-glucoside (Cyn-3-glu) are minor ones. Furthermore, it was detected by UHPLC
ESI MS/MS analysis that although delphinidin-3-sambubioside is the major one, the cyaniding-3-sambubioside is found as the predominant one (Shruthi et al., 2016). The dye can be extracted from the calyces by using small samples of fresh or dried calyces. The fresh calyces can be processed raw by first soaking dried calyces in water overnight. Extract of Roselle calyces did show the qualities of a good substitute for amaranth, to act as a coloring agent for pediatric oral medicine formulations, once it is buffered at pH 5.0 and protected against high temperatures and light (Shruthi et al., 2016). However, as a result of color diminishing and off-flavor development which limits the shelf life of commercial products containing anthocyanins, it tends to restrict the utilization of anthocyanins for specific applications. Therefore the determination of the antioxidant properties, as well as studying the changes in the color and anthocyanin contents of the Roselle extract under different pH situations, is appropriate (Wu, Yang, & Chiang, 2018). The total yield of pigment product was 49.6 mg/g for dried calyces, whereas the Roselle anthocyanins content in the pigment product represented 4.85% (Chang et al., 2012). Shruthi et al. (2016) reported that anthocyanins are highly unstable molecules in food medium. Indeed, different factors strongly affect the color stability of anthocyanins. They include pH, solvents, temperature, anthocyanin concentration and structure, oxygen, light, enzymes, water activity, and other accompanying substances (Arueya & Akomolafe, 2014). It is also important to emphasize that the combination rather than one of the various factors alone drives color stability of anthocyanins. The degradation of enzyme and their interactions with food components such as ascorbic acid, sugars, metal ions, sulfur dioxide, and copigments are also very important in describing color stability of anthocyanins (Abou-Arab, Abu-Salem, & Abou-Arab, 2011). The interaction of all these parameters with food constituents can affect both the formation and stability of anthocyanins (Idham, Muhamad, Mohd Setapar, & Sarmidi, 2012). Light is among the factors that affect anthocyanin stability. From an experiment, about 30% of anthocyanin in grape juice samples was depleted when pigments were exposed to dark condition for 135 days at 20 C (Palamidis & Markakis, 1975). However, when the same samples were held at the same temperature and storage time, light affected more than 50% of the total pigments. This information is beneficial, creating the appropriate conditions to maintain anthocyanin properties in food products. Out of these factors, pH is one of the major factors that significantly influence the pigment color variants and stability of anthocyanin contents. However, the interaction between temperature and pH showed that anthocyanins demonstrated a certain degree of heat resistance and produced a more stable color in acidic media at low pH values than in alkaline solutions (Aishah, Nursabrina, Noriham, Norizzah, & Shahrimi, 2013). Therefore the positive degree of heat resistance in acidic solutions is a proof of Roselle anthocyanins as a potential for application in acidic beverage, including nutritional supplement and pigment in drinks (Wu

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et al., 2018). In the same study, the authors reported that at higher temperatures under low-acid conditions, a higher degradation rate of Roselle anthocyanins was detected. Temperature effect has demonstrated anthocyanin stability in soft drinks, where increased storage temperature significantly accelerated the loss of colorants in soft drinks (Palamidis & Markakis, 1975). Similarly, Maccarone, Maccarrone, and Rapisarda (1985) reported that anthocyanins in red-orange juice deteriorated rapidly when the temperature was increased from 15 C to 35 C during a storage period of 15 days. The susceptibility of anthocyanins to changes in pH is attributed to the ionic nature of anthocyanin. Anthocyanins reportedly occur in four stability forms: the quinonoidal base, flavium cation, carbinol (pseudo-base), and chalcone (Shruthi et al., 2016). Amalia and Afnani (2013) described clearly in their paper the mechanism of action in the four existing equilibrium forms of anthocyanins and their corresponding outcomes. The very rapid degradation of anthocyanins results in color changes, and ultimately, quality and appearance are affected. So, the provision of appropriate acid and temperature conditions prove that Roselle anthocyanins are good sources of beneficial ingredients and color. However, since anthocyanin is somewhat unstable, its high reactivity makes them entirely liable to degradation. For that, they end up forming colorless or undesirable brown-colored compounds during the process of extraction and storage (Durst & Wrolstad, 2001). Therefore one of the recommended approaches to stabilize anthocyanins is encapsulation. Encapsulation enables the preservation of light and heat-labile molecules to maintain their stability and further increase shelf life and bio-activity. As a highly specialized method with relatively affordable cost, various encapsulations are rapidly growing technologies. However, only a few available encapsulated anthocyanins have been evaluated. The use of spray drying method for the application of different coating materials, gelation (polymers as sodium alginate, pectin, curdlan, and glucan), and lyophilization, have been reported (Santos, Albarelli, Beppu, & Meireles, 2013). In other words, the application of microencapsulation has been beneficial in the food industry. It protects sensitive food ingredients during storage, to mask or preserve aromas and flavors, to preserve food against nutritional losses, or even to add nutritive materials to food after processing. It has been estimated that 13,000
14,000 kg of fresh calyces will produce 1800
2000 kg dried calyces plus 2500
3000 kg quantity of seeds per hectare may be attained. Then, the dye recovery from this same quantity of calyces is 1.97%. The edible fixed oil content in the seeds is between 17% and 20%, thus similar in its properties to cottonseed oil (Shruthi et al., 2016). Kim, Lim, Chon, Song, and Seo (2019) in an experiment demonstrated that milk blended successfully with 1.0% Roselle powder, 2.0% of the powder into yogurt, and 3.0% of the Roselle powder mixed well with Kefir. In this study, the taste, flavor, and color of these products were significantly improved for the overall consumer acceptability. The anthocyanin contents of the beverage products have an influence on nutritional loss of those beverages and also their organoleptic characteristics, thus leading to declined antioxidant ability and other effects on bioactivities.

9.2 Use of whole Roselle plant constituents in food

Investigations have shown that Roselle has potential as a feed additive in the feed. Like other herb ingredients, Roselle has potential as a feed additive in the diet of chicken broilers. According to Al-Nasrawi (2013), the intake of Roselle flower supplemented feed by broilers improved their productive performance, hematological and biochemical values. Roselle flower supplemented feed formulated at 1 g/kg can attain acceptable optimum broilers performance; thus body weight, body weight gain, feed consumption, and enhancement in feed conversion ratio significantly (P , .05) increased. Hamodi and Al-K (2011) confirmed that the observed improvement in the measured parameters might be occasioned by the active compounds (anthocyanin and protocatechuic) in Roselle as well as available vitamin C content. The presence of these active compounds set a positive effect on each cell activity and the upsurge in O2 intake; consequently, the thyroid gland is stimulated in the performance of a significant role in metabolism. Furthermore, the positive relationship that was noticed between vitamin C increased phenylalanine and tyrosine metabolism. It also caused the synthesis of primary amino acid in thyroid hormones and maintained secretion of growth hormone due to the increase in basal metabolism (Rosalizan, Rohani, Khatijah, & Shukri, 2008).

9.2.2 Leaves The young leaves and tender stems of Roselle are eaten raw in salads or cooked singly as greens or together with other vegetables and with meat or fish. The leaves are also added to curries as a seasoning. In Africa, India, and Mexico the juice extract of the leaves of Roselle had good treatment effect on various ailments including cholerectic, diuretic, febrifuge, and hypotension, whereas traditionally, it was used for stimulating the intestinal peristalsis and for the reduction of the blood’s viscosity (Da-Costa-Rocha et al., 2014; Riaz & Chopra, 2018). Extract from Roselle leaves can be used for a lotion that is used on sores and wounds (Qi, Chin, Malekian, Berhane, & Gager, 2005). The leaves are used as a source of mucilage in pharmaceutical and cosmetic products. The use of ethanol extract from the dried leaves has the potential to reduce aflatoxin formation (El Shayeb & Mabrouk, 1984).

9.2.3 Roselle seeds Roselle seeds are valuable nutritional food resource of high protein content, calories as well as a large quantity of fiber and useful micronutrients (Akanbi, Olaniyan, Togun, Ilupeju, & Olaniran, 2009). The seeds can be roasted and milled into powder for products like soups and sauces. Besides, the roasted seeds can be used as a substitute for coffee (Mohamed et al., 2012; Qi et al., 2005). In some African countries (including Burkina Faso, Mali, Niger, Nigeria, Cameroon, and Sudan), the seeds of Roselle are subjected to fermentation process to obtain a type of paste used as food condiment (Parkouda, Diawara, & Ouoba, 2008). In Central Burkina Faso, this food condiment is most popular, and the rural people mostly use it in a sauce, eaten with cereals pasta. The processed Roselle food

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condiment is similar to fermented Parkia biglobosa seeds (Atta et al., 2011). It is a cheap functional food serving as both antioxidants and probiotics (Mohamadou, Mbofung, & Thouvenot, 2007). The intake of the fermented Roselle seeds can cure high blood pressure, diarrhea, and rubella or is used as an antiseptic. The flour produced from Roselle seeds is a potential ingredient for food applications; thus it can be used as replacement for production of cookies (Juhari & Petersen, 2018; Nyam, Leao, Tan, & Long, 2014). The reasons are that the seeds contain high levels of protein (13%
35.4%), dietary fiber (18.3%
42.6%), lipid (17.4%
29.6%), and minerals (23.7
128 mg Ca, 596
672 mg P, 2.08
4.0 mg Zn, 0.21
3.1 mg Cu, 26.34
396 mg Mg, 0.08
0.18 mg Cr, and 0.36
0.51 mg riboflavin) (Dhar, Kar, Ojha, Pandey, & Mitra, 2015; El-Adawy & Khalil, 1994; Hainida, Ismail, Hashim, Mohd.-Esa, & Zakiah, 2008; Nyam et al., 2014; Rao, 1996; Tounkara, Amza, Lagnika, Le, & Shi, 2013). In addition, the flour has shown good water and oil absorption capacity, which are good baking qualities for food products development. Seeds of the Roselle exhibit a low content of free fatty acids (suggesting low enzymatic hydrolysis), hence appropriate raw material for preparation of food products (Eltayeib & Elaziz, 2014). The decoction obtained from the seeds is used to improve lactation in situations of poor milk production and maternal mortality in parts of Nigeria (Gaya, Mohammad, Suleiman, Maje, & Adekunle, 2009). Roselle seeds can be used as feed meal for fish and other domestic animals (Mukhtar, 2007). The seeds have been used successfully as additives or supplement in feed meals for broilers chicken production (Al-Nasrawi, 2013; Hamodi & Al-K, 2011; Salih & Abdel Wahab, 1990). Also, Roselle seed oil extract has proven to have in vitro inhibitory effect on Bacillus anthracis and Staphylococcus albums (Gangrade, Mishra, & Kaushal, 1979). Literature on the health values of the Roselle plant roots is lacking, though it has been found to be aphrodisiac (Mungole & Chaturvedi, 2011). It is one of the health and nutritional claims with limited studies to corroborate it. Even more, there is a need for scientific evidence to authenticate its application in the processing of functional food products.

9.3 Other functional food applications Roselle extracts The Roselle calyces residue (RCR) has been incorporated successfully into crackers. The results showed that 5% of the Roselle residue gave the highest dietary fiber, which increased from 3.36% to 8.17%. Other enhanced parameters like increased ash content, increased phenols content from 5.99 to 17.57 mg/g, and increased total flavonoid content from 49.36 to 104.63 mg/g of crackers were observed (Ahmed & Abozed, 2015). The same treatment (addition of 5% RCR) resulted in DPPH radical scavenging activity up by twofold. RCR incorporated into the crackers contributed to darker L values than the less or untreated containing samples. The addition of the RCR to the cracker formula gave it a pleasant

9.4 Conclusion

flavor and consumer acceptability. Therefore the RCR that was discarded after the extraction of beverage causing environmental pollution is now a useful raw material in cracker production. RCR is a potential functional food ingredient rich in high fiber content and antioxidants activity that can be converted into flour for the purpose of food production including baked products. Addition of 0.2% and 0.4% of Roselle calyces to yogurt contributed to enhanced color, increased the sensory parameters, and extended the shelf life for 21 days (Biomy, 2017). Recently, a study on probiotic goat-milk yogurt enhanced with Roselle extract gave the highest antidiabetic effectiveness of 37% α-glucosidase inhibition. Roselle extract added to the probiotic yogurt reduced the inhibitory activity during 15 days of cold storage (4 C). The inhibition activity can be compared with acarbose at 0.1
0.5 ppm concentration. The evaluation of the physical, chemical, and microbiological characteristics of yogurt at 15 days of cold storage showed that it was wholesome consumption (Wihansah, Arief, & Batubara, 2018). The presence of Roselle extract increased the antioxidant component and developed the functional properties of yogurt, making it a healthy dairy product (Biomy, 2017). The consumption of yogurt improves the gastrointestinal functions (Heyman, 2000) and reduces hypertension by lowering the level of cholesterol (Taylor & Williams, 1998). The addition of a natural agent such as the Roselle to yogurt gives it the functional properties as well as increases food variety. The Roselle has anthocyanin and organic acids contents (Al-Wandawi, 2015), and the availability of high phenolic compounds and flavonoids in such plant-based foods has been linked to intestinal α-glucosidase inhibitory activities (Abou-Arab et al., 2011). The addition of Lactobacillus acidophilus IIA-2B4 and Roselle extract to goat-milk yogurt had been reported to have antihypertensive, antimicrobial, and antioxidant activities (Arief, Budiman, Hanifah, & Soenarno, 2016; Hanifah, Arief, & Budiman, 2016; Suharto, Arief, & Taufik, 2016). Therefore Roselle has high antioxidant activity, makes it a useful ingredient to fortify or supplement regularly consumed foods to fight fatal diseases.

9.4 Conclusion Many research works have been focused on Roselle and its extracts consisting of functional properties for the development of new products with additional nutritious characteristics to offer health benefits to various consumers. Multiple uses of Roselle appear to be excellent and promising sources, including functional foods, food colorants, food condiments, and feed supplements. Useful food products from Roselle may provide health benefits to consumers for its substantial contribution of phytochemicals. In this sense, one of the main challenges that companies face today is the development of new value-added products to meet the consumer’s demands. The vast demand for these products by consumers worldwide will promote private enterprise opportunities and improve the socioeconomic conditions of the farming communities of product origin.

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Hainida, E., Ismail, A., Hashim, N., Mohd.-Esa, N., & Zakiah, A. (2008). Effects of defatted dried Roselle (Hibiscus sabdariffa L.) seed powder on lipid profiles of hypercholesterolemia rats. Journal of the Science of Food and Agriculture, 88(6), 1043
1050. Hamodi, S. J., & Al-K, F. M. (2011). Compared between anise seeds (Pimpinella anisum L.) and Roselle flowers (Hibiscus sabdariffa) by their affected on production performance of broiler. Advances in Environmental Biology, 461
465. Hanifah, R., Arief, I., & Budiman, C. (2016). Antimicrobial activity of goat milk yoghurt with addition of a probiotic Lactobacillus acidophilus IIA-2B4 and Roselle (Hibiscus sabdariffa L.) extract. International Food Research Journal, 23(6). Heyman, M. (2000). Effect of lactic acid bacteria on diarrheal diseases. Journal of the American College of Nutrition, 19(Suppl. 2), 137S
146S. Hirunpanich, V., Utaipat, A., Morales, N. P., Bunyapraphatsara, N., Sato, H., Herunsale, A., & Suthisisang, C. (2006). Hypocholesterolemic and antioxidant effects of aqueous extracts from the dried calyx of Hibiscus sabdariffa L. in hypercholesterolemic rats. Journal of Ethnopharmacology, 103(2), 252
260. Idham, Z., Muhamad, I. I., Mohd Setapar, S. H., & Sarmidi, M. R. (2012). Effect of thermal processes on Roselle anthocyanins encapsulated in different polymer matrices. Journal of Food Processing and Preservation, 36(2), 176
184. Jabeur, I., Pereira, E., Barros, L., Calhelha, R. C., Sokovi´c, M., Oliveira, M. B. P., & Ferreira, I. C. (2017). Hibiscus sabdariffa L. as a source of nutrients, bioactive compounds and colouring agents. Food Research International, 100, 717
723. Juhari, N. H., Bredie, W. L., Toldam-Andersen, T. B., & Petersen, M. A. (2018). Characterization of Roselle calyx from different geographical origins. Food Research International, 112, 378
389. Juhari, N. H., & Petersen, M. A. (2018). Physicochemical properties and oxidative storage stability of milled Roselle (Hibiscus sabdariffa L.) seeds. Molecules (Basel, Switzerland), 23(2), 385. Khalid, H., Abdalla, W. E., Abdelgadir, H., Opatz, T., & Efferth, T. (2012). Gems from traditional north-African medicine: Medicinal and aromatic plants from Sudan. Natural Products and Bioprospecting, 2(3), 92
103. Kim, S.-H., Lim, H.-W., Chon, J.-W., Song, K.-Y., & Seo, K.-H. (2019). Sensory profiles of dairy products supplemented with Hibiscus sabdariffa Linnaeus (Roselle) powder: A preliminary study. Journal of Milk Science and Biotechnology, 37(4), 247
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CHAPTER

Nutritional properties and feeding values of Hibiscus sabdariffa and their products

10

Maurice Tibiru Apaliya1, Emmanuel Kwaw1, Gustav Komla Mahunu2, Mildred Osei-Kwarteng3, Richard Osae1 and Michael Azirigo4 1

Department of Food Science and Postharvest Technology, Faculty of Applied Sciences, Cape Coast Technical University, Cape Coast, Ghana 2 Department of Food Science and Technology, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana 3 Department of Horticulture, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana 4 Department of Agriculture for Social Change, Regentropfen College of Applied Sciences, Bongo, Ghana

Chapter Outline 10.1 Introduction ...............................................................................................137 10.2 Nutritional properties of Hibiscus sabdariffa ................................................138 10.2.1 Nutritional composition ...........................................................138 10.2.2 Organic acids ..........................................................................142 10.3 Feeding values of Hibiscus sabdariffa ..........................................................145 10.3.1 Health benefits .......................................................................145 10.3.2 Uses and value .......................................................................147 10.4 Products of Hibiscus sabdariffa ...................................................................148 References ..........................................................................................................150

10.1 Introduction Hibiscus sabdariffa commonly known as “Roselle” or “red sorrel” is a member of Malvaceae family. It is a medicinal plant with a global fame and is estimated to have more than 300 species distributed in tropical and subtropical regions around the world (Singh, Khan, & Hailemariam, 2017). Many parts of H. sabdariffa such as the leaves, seeds, fruits, and roots are used in various foods as well as in herbal Roselle (Hibiscus sabdariffa). DOI: https://doi.org/10.1016/B978-0-12-822100-6.00006-9 © 2021 Elsevier Inc. All rights reserved.

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medicine as a potential nonpharmacological treatment. It is a plant that is of increasing interest due to its enormous nutritional, health, and cosmetic benefits. The fleshy calyx of H. sabdariffa is used in many countries as food or as food ingredients such as jellies, beverages, sirups, cakes, puddings, and wines (Christian, Nair, & Jackson, 2006). The young leaves and succulent stems of Roselle are eaten raw in salads or cooked as greens alone or in combination with other vegetables and/or with meat. They are also added to curries as seasoning. The seeds are high in protein, can be roasted and ground into a powder then used in soups and sauces (Qi, Chin, Malekian, Berhane, & Gager, 2005). The roasted seeds can be used as a substitute for coffee. The red calyx is the most commonly used and is distinctively different from the green and white calyces due to its high concentration of anthocyanin. Cyanidin-3-sambubioside and delphinidin-3sambubioside are the major anthocyanin found in red calyx. The extracts from Roselle are used in the treatment of crucial medical problems such as cancer, helminthic disease, and many cardiovascular disorders (Singh et al., 2017). The plant can also be used as an antioxidant in the management of obesity. Roselle is primarily cultivated commercially for its calyx, which comes in three types: red, green, and dark red. Roselle is also rich in minerals, organic acids, amino acids, vitamin C, carotene, and total sugar in its calyx, seeds, and leaves at varying levels depending on the cultivar and geographical area in which it is grown (Cisse et al., 2009). The flavor of Roselle or hibiscus is a combination of sweet and tart, similar to cranberry (Plotto, 1999).

10.2 Nutritional properties of Hibiscus sabdariffa 10.2.1 Nutritional composition The nutritional composition of fresh and dried Roselle varies among studies, probably due to genetic, different varieties, ecological, environmental, and harvest conditions. H. sabdariffa L. has been used traditionally as food such as herbal drinks, beverages, flavoring agent, and herbal medicine (Da-Costa-Rocha, Bonnlaender, Sievers, Pischel, & Heinrich, 2014). Many parts of H. sabdariffa L. such as the seeds, fruits, leaves, and roots are used in several foods. Proximate analysis of the nutritional composition of Roselle plant showed that carbohydrate content was 68.7%, crude fiber 14.6%, and ash content 12.2% (Luvonga, Njoroge, Makokha, & Ngunjiri, 2010). Roselle plant is also observed to be rich in minerals especially (magnesium, and potassium) vitamins (niacin, ascorbic acid, and pyridoxine) in appreciable amounts.

10.2.1.1 Carbohydrates The carbohydrate level in Roselle like other nutrients varies according to species, the part of the plant, as well as the influence of environmental factors. For instance, carbohydrate levels were found at 69.11% and 69.39% for the white and

10.2 Nutritional properties of Hibiscus sabdariffa

red calyces, respectively (Ahmed, Satti, & Eltahir, 2019). Early studies recounted that H. sabdariffa contains protein (1.9 g/100 g), fat (0.1 g/100 g), carbohydrates (12.3 g/100 g), and fiber (2.3 g/100 g). In addition, the following levels (57.16% and 61.55%) of carbohydrates were found in red and white calyces. According to a report by Singh et al. (2017), carbohydrate levels of 10.2, 25.5, and 8.7 g were observed in the calyces, seeds, and leaves of H. sabdariffa L., respectively. A similar research by Luvonga et al. (2010) revealed that Roselle calyces have a high proportion of carbohydrates which leads credence to the study of (Babalola, 2000; Ojokoh, 2010) who also reported that Roselle calyces contain high carbohydrate contents. Jabeur et al. (2017) in a study recorded the carbohydrate level (87 g/100 g) dw of the Roselle and it was thus the most abundant macronutrient. The nutritional analysis of Roselle calyces by proximate method found that the carbohydrate content (68.7%) was highest followed by crude fiber (14.6%) (Luvonga et al., 2010).

10.2.1.2 Protein and amino acid Many researches have been performed to assess the potential of Roselle as a source of protein and amino acids. Amino acids can be grouped into essential and nonessential amino acids. The nonessentials ones are those that can be synthesis by the body. The Roselle seeds are high in protein, can be roasted and ground into a powder, and used in many dishes (Qi et al., 2005). Roselle calyces have been reported to have high protein content and most of the essential amino acids except tryptophan (Cid-Ortega & Guerrero-Beltra´n, 2015). Nyam, Leao, Tan, and Long (2014) reported that H. sabdariffa L. seed is rich in protein content and other micronutrients, thus serves as a valuable food resource. Besides that, Halimatul, Amin, Mohd.-Esa, Nawalyah, and Siti Muskinah (2007) also confirmed that the high protein quality is found in boiled Roselle seeds. The seed of Roselle contains 27.78% of crude protein (Da-Costa-Rocha et al., 2014).

10.2.1.3 Lipids Lipids are a family of organic compounds that are mostly insoluble in water. They are made up of oils and fats, they have molecules that yield high energy and have a chemical composition mainly of hydrogen, carbon, and oxygen. Hibiscus seeds are a rich source of lipid-soluble antioxidants, particularly γ-tocopherol (Mohamed, Fernandez, Pineda, & Aguilar, 2007). In addition, Tounkara, Amza, Lagnika, Le, and Shi (2013) reported that hibiscus seeds are a good source of lipids. Another finding by Hainida, Ismail, Hashim, Mohd.-Esa, and Zakiah (2008) found that Roselle seeds from Malaysia were composed of 13.0% lipids. The seeds that are mostly considered as a by-product have a lot of nutrients. Raw freeze-dried, sun-dried, and boiled sundried seeds were observed to contained lipids; 2.3%, 13.0%, and 4.0% lipid, respectively (Hainida, Amin, Normah, & Esa, 2008). A study conducted by Nyam et al. (2014) reported that crude lipid content in the seed was significantly higher (P , .05) than that reported by Nyam, Tan, Lai, Long, and Man (2009). It has a considerable amount of fats or lipids that are of benefits to human health.

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Table 10.1 Mineral composition in mg/100 g of Hibiscus sabdariffa calyx. Constituents

Dry purple calyx

Dry red calyx

Dry yellow calyx

Dry green calyx

Calcium Manganese Iron Phosphorus Sodium Zinc Copper B carotene Thiamine Riboflavin Ascorbic acid Potassium Magnesium Nickel

280.00 195.40 58.80 34.50 80.70 81.40 ND ND ND ND 31.24 160.50 ND ND

3.00 1.00 833.00 22.00 15.33 1.17 0.70 285.29 24.67 0.95 53 ND ND ND

3.00 1.07 800.67 23.33 16.01 1.37 0.78 281.28 Trace 0.96 56.83 ND ND ND

21.58 ND 3.37 15.05 48.19 16.28 ND ND ND ND 12.50 49.59 47.54 3.57

10.2.1.4 Minerals The Roselle plant is found to be rich in minerals especially magnesium and potassium. Variable content of minerals were observed in Roselle samples analyzed, indicating that the type of soil influences ash and mineral content within the same species (Carvajal-Zarrabal et al., 2012). In a study conducted by Ahmed et al. (2019), it was found that calcium level in red calyces was 25 mg/100 g while that of the white calyces was 25.5 mg/100 g. In the same study, the red calyces were observed to contain 55.5 mg/100 g of iron, while that of the white calyces contain 30.5 mg/100 g. Similar results (25 and 20 mg/100 g) were reported for red and white calyces (Ahmed et al., 2019). The Roselle calyx is a good source of anthocyanin, ascorbic acid, malic acid, and minerals, especially Fe and Ca, but low in glucose (Jung, Kim, & Joo, 2013). A blend of tropical fruits with Roselle extract have enhanced mineral components and antioxidant properties as Roselle is a good source of iron, calcium, and magnesium (Anel, Thokchom, Subapriya, Jenita Thokchom, & Singh, 2016). Table 10.1 and 10.2 show the mineral composition H. sabdariffa L. calyces and seed, respectively. Furthermore, in a study conducted by Mohamed, Sulaiman, and Dahab (2012), it was revealed that hibiscus flower tea recorded 7% mineral salts.

10.2.1.5 Vitamins Vitamins are substances needed by the body to grow and develop normally. In the last decade, there has been increasing interest in the consumption of natural foods, specifically those made from vegetables. This increasing demand is due to advertisements and scientific research which showed the benefits that these foods

10.2 Nutritional properties of Hibiscus sabdariffa

Table 10.2 Mineral composition in mg/kg of raw and processed Hibiscus sabdariffa seeds cultivated in Cameroon (Ghislain et al., 2011). Sample

Ca

Mg

K

Na

P

Fe

Zn

D2I D2II D3I D3II D4I D4II D5II D6II

1750 1920 1054 1160 1710 1860 1260 1262

1985 2083 1670 1989 1895 2090 1898 1850

30.52 272.7 26.45 232.6 31.8 272.7 205.45 206.9

22 14 21 15 24 18 21 20

10.58 35.82 20.73 80.60 90.78 18.92 40.52 42.03

137.3 147.5 145.02 169.22 140.01 152.5 148.7 149.03

63.5 76.1 21.3 10.13 46.91 52.6 69.8 70.11

D2I and D2II 5 raw Roselle seeds, D3I and D3II 5 boiled Roselle seeds, D4I and D4II 5 toasted Roselle seeds, D5II 5 toasted for 7 min after soaking for 12 h, D6II 5 toasted for 14 min after soaking for 12 h. I 5 agro ecological area I (Adamaoua region). II 5 agro ecological area II (West region).

contribute to health (Falk, 2004; Hasler, 2002). These foods known as functional foods contain significant levels of active biological components, mainly anthocyanins, flavonoids, vitamins, and polyphenolic acids, which might provide specific health benefits to humans beyond the traditional nutrients that they are composed of. A study conducted on the juice obtained from the calyces of Roselle revealed that the drink has a high content of vitamin C, antioxidant, and anthocyanin (Abu-Tarboush, Ahmed, & Al Kahtani, 1997; Haruna, 1997; Mohamed et al., 2012). The calyces of Roselle for instance contain nine times vitamin C than Citrus sinensis (Ismail, Ikram, & Nazri, 2008). The leaves and seeds of Roselle have also been reported to be rich in vitamin C (Augstburger et al., 2002). Barker and Pilbeam (2015) reported that every 100 g of fresh calyx contains 0.5 mg vitamin B complex, 2.85 μg vitamin D, 0.04 mg vitamin B1, and 0.6 mg vitamin B2. Essentially, H. sabdariffa L. is noted to have vitamin C (water soluble). Table 10.3 shows water-soluble vitamins in dry and fresh calyces of rosselle before processing.

10.2.1.6 Dietary fiber Roselle seed is a valuable source of food as it has an appreciable amount of dietary fiber (Nyam et al., 2014). Furthermore, according to Hainida, Ismail, et al. (2008), hibiscus seeds contain 18.3% of total dietary fibers. A substantial amount of crude fiber (24.7%) indicated that the Roselle seed could be considered as a good source of dietary fiber (Nyam et al., 2014). Besides, Roselle seed contained a balance proportion of soluble and insoluble fraction of dietary fiber (Hainida, Ismail, et al., 2008). Roselle seed could be regarded as a useful and low-cost source of dietary fiber substitute in dietary supplement or food ingredient. Thus Roselle seed powder has the potential to be used as a source of dietary fiber and can be developed as a functional ingredient in food products. Therefore the

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Table 10.3 Water-soluble vitamins in fresh and dried calyces of Roselle before processing. Constituent vitamins

Fresh calyces (mg/100 g)

Dry calyces (mg/100 g)

Niacin Thiamin Riboflavin Pantothenic Folic acid Ascorbic acid Pyridoxine

3.765 0.177 0.277 0.324 0.122 6.701 1.546

2.644 0.123 0.194 0.227 0.092 4.690 1.080

Source: Adapted from Luvonga, W.A., Njoroge, M., Makokha, A., & Ngunjiri, P. (2010). Chemical characterisation of Hibiscus sabdariffa (Roselle) calyces and evaluation of its functional potential in the food industry. In Paper presented at the JKUAT annual scientific conference proceedings.

consumption of Roselle seed powder will improve the daily intake of dietary fiber and augment the fiber deficit. The edible seeds from pulses are a rich food source of dietary fibers that enhance various health benefits (Tosh & Yada, 2010). The hull of seed has a significant quantity of insoluble dietary fiber, while the inner part of seeds contains soluble dietary fiber. Dietary fiber present in Roselle seeds contributes to physiological and health benefits. Roselle seed powder that is considered a by-product can be recycled as value-added food supplements, which provide advantageous dietary fiber and bioactive compounds. A similar study also revealed that Roselle flower tea was found to have 15% fiber present (Mohamed et al., 2012).

10.2.2 Organic acids Roselle has been reported to be rich in organic acids such as malic, citric, tartaric, and allo-hydroxycitric acids (Singh et al., 2017). The rough look of H. sabdariffa L. fortified sample is due to pores caused by the leaven promoter which was believed to be a result of acidic components (Daramola & Asunni, 2006). A study conducted by Ahmed et al. (2019), on H. sabdariffa extract, revealed the presence of a great variety of organic acids and phenolic compounds such as hibiscus acid hydroxycitric, citric acid, and protocatechuic acid. Extract from the dried leaves of H. sabdariffa L. showed the presence of 4.25% catechin and 28.20% ellagic acid (Lin et al., 2012), while the extract from fresh leaves showed the presence of 24.24% protocatechuic acid, 2.6% catechin, 2.44% gallocatechin, 19.85% caffeic acid, and 27.98% gallocatechin gallate (Yang et al., 2010). Similar findings were reported by Huang et al. (2009). Among these, hibiscus acid is one of the major organic acids in Roselle, which include oxalic acid, tartaric acid, malic acid, and succinic acid, those organic acids compose 15%
30% of the Roselle calyces mass (World Health Organization, 1998). Ascorbic acid is also present

10.2 Nutritional properties of Hibiscus sabdariffa

Table 10.4 Nutritional composition of Hibiscus sabdariffa Organic acid content

Nutrient value (mg/100 g dw)

Oxalic acid Malic acid Shikimic acid Fumaric acid Ascorbic acid

1.81 9.10 0.36 0.04 54

FIGURE 10.1 (A) (2S, 3R)-hydroxycitric acid. (B) Hibiscus acid. (B) Adapted from Da-Costa-Rocha, I., Bonnlaender, B., Sievers, H., Pischel, I., & Heinrich, M. (2014). Hibiscus sabdariffa L.—A phytochemical and pharmacological review. Food Chemistry, 165, 424
443.

in H. sabdariffa L.; however, its content differs greatly between dried [260
280 mg/100 g (Ismail et al., 2008)] and fresh calyces [(6.7
14 mg/100 g) (Ismail et al., 2008; Morton, 1987)]. The amount of ascorbic acid in the dried calyx being much higher than the ones previously reported in the literature. Table 10.4 shows organic acid content of H. sabdariffa L. Hydroxycitric acid present in the calyx has an additional hydroxyl group at the second carbon of citric acid. Hydroxycitric acid has four stereoisomers—(2S, 3S), (2S, 3R), (2R, 3R), and (2R, 3S)—and their lactone forms. The major stereoisomers found in the Roselle is the (2S, 3R)-hydroxycitric acid (Hida, Yamada, & Yamada, 2007). It is the major organic acid found in the calyces of Roselle (Fig. 10.1A). Hibiscus acid compromises a citric acid moiety with an additional hydroxyl group at the second carbon and has two diastereomers as are result of the existence of two chiral centers in the molecule (Eggensperger & Wilker, 1996). It is the lactone form of (1)-allo-hydroxycitric acid (Fig. 10.1B).

10.2.2.1 Flavonoids H. sabdariffa L. contains polyphenols of the flavonol and flavanol type in simple or polymerized form. The principal compounds present in Roselle calyx extracts are flavonoids and anthocyanins, which act as antioxidants and they have no

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mutagenic or toxic activity (Cid-Ortega & Guerrero-Beltra´n, 2015). In a similar study, Puro et al. (2014) reported that the extract contained flavonoids, saponins, cardiac glycosides, and alkaloids. Other flavonoid components that exist in H. sabdariffa calyx include anthocyanins as cyanidin-3-rutinoside, delphinidin, delphinidin-3-monoglucoside, cyanidin-3-monoglucoside, cyanidin-3-sambubioside, cyanidin-3,5-diglucoside; the flavonol glycosides hibiscetin-3-monoglucoside, gossypetin-3-glucoside, gossypetin-7-glucoside, gossypetin-8-glucoside, and sabdaritrin (Hirunpanich et al., 2005). The presence of these flavonol glycosides was low, with hibiscitrin being the major compound followed by gossypitrin and sabdaritrin (Rao & Seshadri, 1942). According to Mishra, Shukla, Jain, and Kumar (1999), a number of compounds have been isolated and characterized from Roselle such as flavonoids, anthocyanidins, steroids, triterpenoids, and alkaloids. Furthermore, Qi et al. (2005) reported that the red calyces from Roselle contain antioxidants such as flavonoids, gossypetine, hibiscetine, and sabdaretine. The leaves of Roselle contains beta-sitosterol-beta-D-galactoside (Osman, Younes, & Mokhtar, 1975) and from the seeds ergosterol (Salama & Ibrahim, 1979) were reported. Ergosterol and b-sitosterol were also reported in Roselle extracts (Mckay, 2009).

10.2.2.2 Anthocyanin Anthocyanins have a long history as a human diet due to their positive health benefits. Anthocyanins are a group of flavonoid derivatives and natural occurring pigments present in dried flowers of H. sabdariffa and their color varies depending on the pH (Da-Costa-Rocha et al., 2014). Anthocyanin is one form of flavonoid constituents that can be found in large amounts in Roselle calyces (Tsai, McIntosh, Pearce, Camden, & Jordan, 2002; Wang et al., 2000). Again, Aziz, Gad, and Badran (2007) reported that H. sabdariffa L. is the primary commercially available food colorant source of anthocyanin. Roselle calyces contain hibiscin and gossypetin which are the two types of anthocyanin natural compounds used for coloring liquors and sirups. Anthocyanin pigments from H. sabdariffa L., calyces are suitable for use as natural food coloring agents. Anthocyanins are universal plant colorants that are responsible for the orange, scarlet, pink, red, violet, blue, and mauve color of fruit and flower colors of higher plants. A study conducted by Wong, Yusof, Ghazali, and Man (2002) revealed a concentration of total anthocyanins (2520 6 50 mg/100 g) of fine powder of Roselle (expressed as delphinidin-3-glucoside). They also indicated that through high-performance liquid chromatography analysis, delphinidin-3-sambubioside (71.4%) and cyanidin-3-sambubioside (26.6%) are the major anthocyanins present in Roselle calyces. In a similar study, Galicia-Flores, Salinas-Moreno, EspinozaGarc´ıa, and Sa´nchez-Feria (2008) reported a total anthocyanins content of between 364.98 and 606.67 mg/100 g dry powdered sample extracted using methanol mixed with 1% trifluoroacetic acid and between 172.58 and 296.99 mg/100 g of dry calyces extracted using distilled water. On the other hand, Azza, Ferial, and Esmat (2011) reported total anthocyanins (622.91 6 2.0 mg cyaniding-3glucoside/100 g) of dry sample using the colorimetric method of analysis.

10.3 Feeding values of Hibiscus sabdariffa

10.3 Feeding values of Hibiscus sabdariffa It is absolutely important to know the feeding value available nutrients and metabolizable energy content of Roselle to obtain a balance diet. Feeding value and nutrient content of Roselle change depending on the stage of maturity, fertilizer application, soil conditions, climate, method of processing, etc. There is no literature that show the basic feeding values of Roselle grown in different regions across the globe. Well-being, convenience, and tolerance continue to dictate the eating habits of modern-day society. Despite the common knowledge on the relation between health and dietary fiber, a big gap exists between recommended daily intake and mean daily intake (Handa, Goomer, & Siddhu, 2012).

10.3.1 Health benefits H. sabdariffa, which belongs to the family Malvaceae, is a well-known medicinal plant with a global fame (Mohagheghi, Maghsoud, Khashayar, & Ghazi-Khansari, 2011). Roselle is an astringent, aromatic herb that is commonly used in the tropics (Islam, 2019). The plant is said to possess diuretic effects; hence, it helps to lower antiscorbutic and fever. Presently, Roselle is drawing the attention of the food and pharmaceutical industries who feel it may have exploitable potentials as a natural food commodity for herbal medicine and as a colorant agent to substitute synthetic dyes. In Africa, India, and Mexico, extracts of the calyces and leaves are traditionally used for their cholerectic, diuretic, hypotensive, and febrifugal effects, decreasing the viscosity of the blood and stimulating intestinal peristalsis. The leaves and flowers are recommended as a tonic for proper kidney function and for intestinal digestion. Roselle plant is used as folk medicine in the treatment of bilious conditions, abscesses, cancer, debility, cough, dysuria, dyspepsia, fever, scurvy, heart ailments, hangover, hypertension, neurosis, and strangury (Nnam & Onyeke, 2003). The extracts from Roselle act as anticancer and may help to reduce cardiovascular and chronic diseases including dyslipidemia, diabetes mellitus, and hypertension (Cid-Ortega & Guerrero-Beltra´n, 2015). According to Hirunpanich et al. (2006), Roselle is used as a traditional medicine for the treatment of kidney and urinary bladder stones. In addition, research on humans proved that the consumption of Roselle has an antiinflammatory effect (Beltra´nDebo´n et al., 2010). This could be due to the activity of some compounds, mainly flavonoids and anthocyanins, found as natural antioxidants in Roselle extracts (Cid-Ortega & Guerrero-Beltra´n, 2015). Numerous researchers have pointed out that Roselle and its extracts possess functional properties from where advances can be made to develop new products with more nutritious features that may provide health benefits to consumers (Ahirwar & Shakya, 2015; Ahmed, Satti, & Eltahir, 2019; Singh, Khan, & Hailemariam, 2017). Foods from Roselle are known as functional foods, due to the health benefits they provide amidst the presence of phytochemicals (Cid-Ortega & Guerrero-Beltra´n, 2015).

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Roselle is valued for its slight laxative effect, relief during hot weather, ability to increase urination, and treatment of feet cracks, sores, bilious, and wounds (Qi et al., 2005). In Sudan, traditionally Roselle has been used in the treatment of sour throat and healing of wounds (Aziz et al., 2007). Apart from Roselle being reported as a coloring agent, it has bioactive properties such as antiinflammatory, antioxidant, antibacterial, and hepatoprotective potential (Vagiri & Jensen, 2017). Preparations from Roselle calyces have been used to treat nerve and cardiac diseases and also to increase diuresis in Egypt (Da-Costa-Rocha et al., 2014). In Nigeria, according to Gaya, Mohammad, Suleiman, Maje, and Adekunle (2009), the decoction from the seeds is usually used to enhance lactation in cases of poor milk production, maternal mortality, and poor letdown. The seed of Roselle could be utilized in the preparation of bakery products and many food commodities due to its nutritional benefits to human health (Nyam et al., 2014). Roselle is commercially grown as a fiber plant and serves as a jute substitute in making linen, clothing, ropes, and fishing nets, and among others.

10.3.1.1 Antioxidant properties of Hibiscus sabdariffa The calyces of Roselle have been extensively studied and demonstrated to have beneficial health effects as a good source of antioxidants (Da-Costa-Rocha et al., 2014; Islam, 2019; Puro et al., 2014). Antioxidant is a substance that slows or inhibits oxidation reaction, especially in biological materials or within cells, thereby reducing spoilage or preventing damage. The main function of antioxidant is trapping the free radical particularly reactive oxygen species and reactive nitrogen species which are involved in the pathogenesis of several chronic and degenerative diseases such as cardiovascular diseases, inflammation, neurodegenerative diseases, aging-related disorders, and cancer. The calyx of Roselle provides higher levels of antioxidants than traditional sources such as blueberries and raspberries (Juliani et al., 2009). Protocatechuic acid and anthocyanins are some of the chemical constituents present in H. sabdariffa that proved to have robust antioxidant activity. Previous studies indicated that extract obtained from dried Roselle calyx was revealed to be active against rat hepatocytes from tert-butyl hydroperoxide
induced cytotoxicity and genotoxicity through different mechanisms (Liu, Schiff, & Dinesh-Kumar, 2002). According to Lin et al. (2003), hibiscus protocatechuic acid has been demonstrated to prevent the carcinogenic activity of various chemicals in various tissues of rat, including diethylnitrosamine present in the liver. Similar study suggested that one of the mechanisms of protection was associated with the scavenging of free radicals by antioxidant compounds present in H. sabdariffa calyx (Ali, Wabel, & Blunden, 2005). H. sabdariffa calyx possesses antioxidant effects against hypolipidemic and low-density lipoprotein oxidation in vivo assay; however, the mechanisms of action is yet to be investigated (Hirunpanich et al., 2006). The tea from the calyces of Roselle is claimed to be a medicinal drink as a result of its high content of antioxidants, vitamin C, and anthocyanins.

10.3 Feeding values of Hibiscus sabdariffa

(A) Red Roselle calyx

(B) Green Roselle calyx

(C) Dark Roselle calyx

FIGURE 10.2 Types of Roselle calyces.

10.3.1.2 Antimicrobial activity of Hibiscus sabdariffa Roselle is widely used for the treatment of diseases. Roselle plant has been used to treat diseases such as bilious conditions, abscesses, cancer, and coughs. For instance, Puro et al. (2014) reported that Roselle exhibited antibacterial activities against Bacillus stearothermophilus, Staphylococcus aureus, Micrococcus luteus, Clostridium sporogenes, Serratia mascences, Escherichia coli, Bacillus cereus, Klebsiella pneumonia, and Pseudomonas fluorescens. In addition, Roselle extract demonstrated antimicrobial activity against Salmonella enterica, E. coli O157:H7, and Listeria monocytogenes isolated from food, animal, and clinical samples (Fullerton, Khatiwada, Johnson, Davis, & Williams, 2011). In another study, ethanol and aqueous extracts obtained from Roselle calyx and protocatechuic acid showed antibacterial effects against Salmonella typhimurium DT104, L. monocytogenes, E. coli O157:H7, B. cereus, and S. aureus (Chao & Yin, 2009). H. sabdariffa extracts demonstrated antimicrobial and antioxidant activities, indicating that the hydroethanol extract presents not only inhibit lipid peroxidation but also fungi and bacteria. Elmanama, Alyazji, and Abu-Gheneima (2011) applied water and methanol extracts of H. sabdariffa on samples of bacteria and fungi and revealed that antimicrobial effect was observed. From the aforementioned, the antimicrobial features of Roselle calyces could be attributed to the abundance of phytochemicals, such as anthocyanins (delphinidin-3-O-sambubioside, and cyanidin-3-O-sambubioside) and phenolic acids (protocatechuic acid) (Gutie´rrezAlca´ntara et al., 2016; Liu, Tsao, & Yin, 2005). According to Nwaiwu, Mshelia, and Raufu (2012), the crude extracts (200 mg/L) from Roselle seed inhibited the growth of three types of Gram-negative bacteria (Fig. 10.2). The extract showed antimicrobial activity against salmonella, shigella, and enterobacter.

10.3.2 Uses and value Roselle plant (H. sabdariffa) Linne (Malvaceae) has been used in folk medicine as a mild laxative, diuretic, and treatment for cardiac and nerve diseases. The economic interest of Roselle varieties lies in their dried calyces that are used worldwide in the production of beverages, jellies, sauces, liqueurs, wines, and preserves; it

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is also used as a source of natural dye for foods due to the presence of anthocyanins (Juliani et al., 2009). The fresh calyx (the outer whorl of the flower Fig. 10.1) is cooked, is eaten raw in salads, and used as a flavor agent in cakes. The calyx is also used in making soups, pickles, jellies, sauces, and puddings. The calyx is a major source of pectin and citric acid and is used in making jams and jellies. It is a good source of red color and for flavoring herb teas and can be roasted and used as a coffee substitute. The calyx together with a small amount of ginger can be boiled and sweeten with sugar to serve as a refreshing beverage. Tender young leaves and stems can be eaten raw or cooked. The leaves can be used in salads, as a potherb and as a seasoning in curries; they have an acid, rhubarb-like flavor. The dry seed can be ground into a powder and used in oily soups and sauces. The oven-dried seeds have been used as a coffee substitute as aphrodisiac properties. The roots are also edible, however, very fibrous. The seed has 20% oil content (Puro et al., 2014). Roselle calyces have been used in the bakery and confectionery industry due to their rich content of calcium, fiber, and iron. Roselle has been used as herbal tea to treat pyrexia, hypertension, and liver damage, although pharmaceutical components are poorly defined (Hou, Tong, Terahara, Luo, & Fujii, 2005). The leaves are extensively used as animal fodder. In China, the seeds are used for the production of oil and the plant is used for medicine (Plotto, Mazaud, Ro¨ttger, & Steffel, 2004). In the Near East and North Africa, Roselle is used in the food and pharmaceutical industries. Roselle has been reported to have mild laxative effect, ability to increase urination, provide relief during hot weather, and for treatment of cracks in the feet, sores, bilious, and wounds (Qi et al., 2005). Traditionally in Africa, Roselle leaves are used for their, emollient, antimicrobial, antipyretic, sedative properties, diuretic, antihelminthic, and as a soothing cough remedy (Duke, 2002). A study by Gosain et al. (2010) affirmed that ethanolic extract from Roselle leaves significantly exhibited hypolipidemic effect. Roselle plant also serves as an antioxidant and is used to manage obesity. Health wise, Roselle extracts are used to manage different medical conditions such as cancer, cardiovascular problems, and inflammatory diseases (Nkumah, 2015). A study conducted by Yamada, Hida, and Yamada (2007) revealed that hibiscus acid [hibiscus type (2S, 3R)-hydroxycitric acid lactone] has the potency to inhibitor intestinal α-glucosidase and pancreatic α-amylase activities (Yamada et al., 2007).

10.4 Products of Hibiscus sabdariffa The leaves, roots, seeds, and calyx of Roselle are used in the manufacture of both local and international products which are consumed globally. Recently, there has been an increase in demand for Roselle and its derived products due to their functional properties. The products of Roselle consumed globally, varies according to geographical location. Hibiscus tea is caffeine-free herbal tea made out of the dried fruit of Roselle, called calyx. It is red in color and tastes like berries. The Fleshy red calyces of

10.4 Products of Hibiscus sabdariffa

Roselle are commonly used for the production of soft drinks and tonic without alcohol like wine, juice, jam, jelly, and sirup and also dried and brewed into tea and spice. Previous studies have shown that enzymes inhibitors are present in Roselle tea which inhibits the production of amylase. For instance, it has been reported by Da-Costa-Rocha et al. (2014) that a cup of hibiscus tea after meals can reduce the absorption of dietary carbohydrates and assist in weight loss. Some of the products produced from Roselle include Roselle tea, Roselle powder, Roselle tablets, Roselle juice, Roselle extracts, Roselle jam, and Roselle seed oil, among several others (Fig. 10.3).

(A) Roselle jam

(E) Roselle drink

(D) Roselle tea

FIGURE 10.3 Products from Roselle.

(B) Roselle wine

(F) Roselle tablets

(C) Roselle powder

(G) Roselle seed powder

(H) Roselle seed

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6869. Cisse, M., Dornier, M., Sakho, M., Ndiaye, A., Reynes, M., & Sock, O. (2009). Le bissap (Hibiscus sabdariffa L.): Composition et principales utilisations. Fruits, 64(3), 179
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523. Elmanama, A. A., Alyazji, A. A., & Abu-Gheneima, N. (2011). Antibacterial, antifungal and synergistic effect of Lawsonia inermis, Punica granatum and Hibiscus sabdariffa. Annals of AlQuds Medicine, 7. Falk, M. (2004). The impact of regulation on informing consumers about the health promoting properties of functional foods in the USA. Journal of Food Science, 69(5), R143
R145. Fullerton, M., Khatiwada, J., Johnson, J. U., Davis, S., & Williams, L. L. (2011). Determination of antimicrobial activity of sorrel (Hibiscus sabdariffa) on Esherichia coli O157:H7 isolated from food, veterinary, and clinical samples. Journal of Medicinal Food, 14(9), 950
956. Galicia-Flores, L. A., Salinas-Moreno, Y., Espinoza-Garc´ıa, B., & Sa´nchez-Feria, C. (2008). Caracterizacio´n fisicoqu´ımica y actividad antioxidante de extractos de jamaica (Hibiscus sabdariffa L.) nacional e importada. Revista Chapingo. Serie Horticultura, 14(2), 121
129. Gaya, I., Mohammad, O., Suleiman, A., Maje, M., & Adekunle, A. (2009). Toxicological and lactogenic studies on the seeds of Hibiscus sabdariffa Linn (Malvaceae) extract on serum prolactin levels of albino wistar rats. The Internet Journal of Endocrinology, 5 (2), 1
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759. Gosain, S., Ircchiaya, R., Sharma, P. C., Thareja, S., Kalra, A., Deep, A., & Bhardwaj, T. R. (2010). Hypolipidemic effect of ethanolic extract from the leaves of Hibiscus sabdariffa L. in hyperlipidemic rats. Acta Poloniae Pharmaceutica, 67(2), 179
184. Gutie´rrez-Alca´ntara, E., Rangel-Vargas, E., Go´mez-Aldapa, C., Falfan-Cortes, R., Rodr´ıguez-Mar´ın, M., God´ınez-Oviedo, A., . . . Castro-Rosas, J. (2016). Antibacterial effect of Roselle extracts (Hibiscus sabadariffa), sodium hypochlorite and acetic acid against multidrug-resistant Salmonella strains isolated from tomatoes. Letters in Applied Microbiology, 62(2), 177
184. Hainida, E., Ismail, A., Hashim, N., Mohd.-Esa, N., & Zakiah, A. (2008). Effects of defatted dried Roselle (Hibiscus sabdariffa L.) seed powder on lipid profiles of hypercholesterolemia rats. Journal of the Science of Food and Agriculture, 88(6), 1043
1050. Hainida, K. E., Amin, I., Normah, H., & Esa, N. M. (2008). Nutritional and amino acid contents of differently treated Roselle (Hibiscus sabdariffa L.) seeds. Food Chemistry, 111(4), 906
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Halimatul, S., Amin, I., Mohd.-Esa, N., Nawalyah, A., & Siti Muskinah, M. (2007). Protein quality of Roselle (Hibiscus sabdariffa L.) seeds. ASEAN Food Journal, 14(2), 131. Handa, C., Goomer, S., & Siddhu, A. (2012). Physicochemical properties and sensory evaluation of fructoligosaccharide enriched cookies. Journal of Food Science and Technology, 49(2), 192
199. Haruna, A. (1997). Cathartic activity of Soborodo: The aqueous extract of calyx of Hibiscus sabdariffa L. Phytotherapy Research: An International Journal Devoted to Medical and Scientific Research on Plants and Plant Products, 11(4), 307
308. Hasler, C. M. (2002). Functional foods: Benefits, concerns and challenges—A position paper from the American Council on Science and Health. The Journal of Nutrition, 132 (12), 3772
3781. Hida, H., Yamada, T., & Yamada, Y. (2007). Genome shuffling of Streptomyces sp. U121 for improved production of hydroxycitric acid. Applied Microbiology and Biotechnology, 73(6), 1387
1393. Hirunpanich, V., Utaipat, A., Morales, N. P., Bunyapraphatsara, N., Sato, H., Herunsale, A., & Suthisisang, C. (2006). Hypocholesterolemic and antioxidant effects of aqueous extracts from the dried calyx of Hibiscus sabdariffa L. in hypercholesterolemic rats. Journal of Ethnopharmacology, 103(2), 252
260. Hirunpanich, V., Utaipat, A., Morales, N. P., Bunyapraphatsara, N., Sato, H., Herunsalee, A., & Suthisisang, C. (2005). Antioxidant effects of aqueous extracts from dried calyx of Hibiscus sabdariffa Linn (Roselle) in vitro using rat low-density lipoprotein (LDL). Biological and Pharmaceutical Bulletin, 28(3), 481
484. Hou, D.-X., Tong, X., Terahara, N., Luo, D., & Fujii, M. (2005). Delphinidin 3sambubioside, a Hibiscus anthocyanin, induces apoptosis in human leukemia cells through reactive oxygen species-mediated mitochondrial pathway. Archives of Biochemistry and Biophysics, 440(1), 101
109. Huang, C.-N., Chan, K.-C., Lin, W.-T., Su, S.-L., Wang, C.-J., & Peng, C.-H. (2009). Hibiscus sabdariffa inhibits vascular smooth muscle cell proliferation and migration induced by high glucose—A mechanism involves connective tissue growth factor signals. Journal of Agricultural and Food Chemistry, 57(8), 3073
3079. Islam, M. (2019). Food and medicinal values of Roselle (Hibiscus sabdariffa L. Linne Malvaceae) plant parts: A review. Open Journal of Nutrition and Food Science, 1(1), 1003. Ismail, A., Ikram, E. H. K., & Nazri, H. S. M. (2008). Roselle (Hibiscus sabdariffa L.) seeds—Nutritional composition, protein quality and health benefits. Food, 2(1), 1
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Medicinal and therapeutic potential of Roselle (Hibiscus sabdariffa)

11

Muhammad Arslan1, Muhammad Zareef1, Haroon Elrasheid Tahir1, Allah Rakha2, Zou Xiaobo1 and Gustav Komla Mahunu3 1

School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China National Institute of Food Science and Technology, University of Agriculture, Faisalabad, Pakistan 3 Department of Food Science and Technology, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana 2

Chapter Outline 11.1 11.2 11.3 11.4

Introduction ...............................................................................................156 Roselle bioactive compounds and their therapeutic benefits .........................157 Roselle uses in traditional medicine ............................................................158 Medicinal and therapeutic health benefits of Roselle ....................................159 11.4.1 Antihypertensive activity ........................................................159 11.4.2 Anti-inflammatory activity ......................................................159 11.4.3 Antiobesity activity ................................................................161 11.4.4 Antidiabetic activity ..............................................................162 11.4.5 Nephroprotective activity .......................................................164 11.4.6 Hepatoprotective activity .......................................................165 11.4.7 Cardioprotective activity ........................................................167 11.4.8 Renal effects, uricosuric effect, and hyperuricemia ..................169 11.4.9 Treatment of anemia ..............................................................170 11.4.10 Cancer preventive activity ......................................................171 11.4.11 Uses against cadmium poisoning ............................................172 11.5 Future perspectives ....................................................................................174 11.6 Conclusion .................................................................................................175 References ..........................................................................................................175

Roselle (Hibiscus sabdariffa). DOI: https://doi.org/10.1016/B978-0-12-822100-6.00007-0 © 2021 Elsevier Inc. All rights reserved.

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11.1 Introduction Plants play an imperative role in the well-being of human being as they deliver the fundamental needs of mankind, that is, food, shelter, clothing, and medicines. They also provide the basis for centuries-old traditional medicinal systems such as Chinese, Unani, Ayurveda, and so on. A large part of the population in developing countries still relies on medicinal plants to accomplish their basic health requirements. Recently, these traditional alternative medicines have gained the attention of the global population owing to their abundance, less cost, and fewer adverse effects. Over the last few years, plant research has extensively focused on ascertaining the vast potentials of therapeutic plants used in traditional medicinal systems (Bloom, 2018; Roy, Mandal, Panda, Roy, & Subba, 2018). Recently, several therapeutic plants have been studied as a potent phytochemical agent for the management of various diseases; Roselle is one of those plants (Ali, Wabel, & Blunden, 2005; Riaz & Chopra, 2018). It is a superlative crop for developing countries as it is relatively easy to cultivate, can integrate into a multicropping pattern, and is used as fiber and food. In Western Africa, seed powder and leaves of the Roselle are used in daily meals, pharmaceuticals, and food processing industries. Likewise, in China, Roselle plant is used for its therapeutic properties in folk medicines, and its seeds are used for oil extraction (Ali et al., 2005; Riaz & Chopra, 2018). Roselle is an underutilized plant and is a potential source of bioactive compounds. The calyx being a commercially vital part of Roselle contains a large pool of secondary metabolites, phenolic acids, organic acids, flavonoids, and anthocyanins. Previous scientific reports gave credence to its use in traditional medicines in the treatment of hyperlipidemia (Gaffer & Mustafa, 2019), hypertension (Al-Anbaki et al., 2019; Herrera-Arellano, Zamilpa, Jime´nez-Ferrer, Cruz-Herna´ndez, & Clemente-Catonga, 2016), and type-2 diabetes (Adeyemi & Adewole, 2019; NganouGnindjio al., 2019). Besides, it possesses anti-inflammatory (El Bayani et al., 2018; Shen, Zhang, Zhang, & Jiang, 2016), nephroprotective (Al-Qahtani, 2018; Shalgum et al., 2019), hepatoprotective (Ezzat et al., 2016), cancer preventive, and antianemia activities (Al Groom & Al-Kubaisy, 2016; Malacrida et al., 2016; Su et al., 2018). It is hypothesized that these medicinal and therapeutic properties are attributed to higher contents of anthocyanin, polyphenols, and coloring pigments founds in flowers and fruits. The main anthocyanin compounds present in calyx include cyanidin-3sambubioside and delphinidin-3-sambubioside (Maciel et al., 2018). The leaves of Roselle are the major portion of foliage, which are mostly discarded except in sub-Sahara Africa where they are consumed as vegetables in sauces and soups. Recent studies on leaf extract have also reported various in vivo and in vitro bioactivities including antiatherosclerotic, antioxidant, antiproliferation, and antihyperlipidemic (Chen, Lee, Wang, Hsu, & Lin, 2017; Chou, Wang, Chen, & Lin, 2019; Nwibo, Eze, & Okonkwo, 2016; Zhen et al., 2016). This chapter aims to highlight the role of the Roselle bioactive component in health promotion.

11.2 Roselle bioactive compounds and their therapeutic benefits

11.2 Roselle bioactive compounds and their therapeutic benefits The free radicals are atoms or groups of atoms with a free or unpaired electron. Free radicals start a chain reaction to achieve electrochemical stability, which may imbalance body homeostasis and harm the biological macromolecules such as carbohydrates, protein, lipid, and nucleic acid (Elejalde, 2001). Most of these free radicals are the products of normal cell metabolism. Nevertheless, free radicals production may also be triggered by exposure to heavy metals, pesticides, ionizing radiation, or sun rays and metabolism of some exogenous substances (Droge, 2002). Other factors affecting free radicals’ production are linked with inadequate diet, tobacco smoke, limited intake of antioxidants, and exposure to xenobiotics such as carbon tetrachloride, acetaminophen, and chloroform. The aforementioned factors may result in excessive free radicals within the body and enhance the vulnerability to pathological conditions such as hyperlipidemia, cancer, atherosclerosis, and so on (Elejalde, 2001; Lobo, Patil, Phatak, & Chandra, 2010). The main components of therapeutic importance are polyphenols, organic acids, polysaccharides, and flavonoids mainly anthocyanins (Chang, Huang, Hsu, Yang, & Wang, 2005; Christian, Nair, & Jackson, 2006; Murkovic, Toplak, Adam, & Pfannhauser, 2000). The dried extract of calyces is known to have substantial concentrations of organic acid (tartaric acid, citric acid, oxalic acid, ascorbic acid, hibiscus acid, and maleic acid) in addition to anthocyanins, phytosterols, watersoluble antioxidants, and polyphenols (Mahadevan & Kamboj, 2009). These bioactive compounds along with organic acids have significant free radical scavenging activity (Luvonga, Njoroge, Makokha, & Ngunjiri, 2010). Thus the valuable health benefits attributed to Roselle are the consequence of its rich phytochemistry. Polyphenols are a heterogeneous group of molecules having various phenol groups in their structure. Recent scientific reports support the antioxidant capacities of polyphenols, though their antioxidant properties rely on their absorption and bioavailability (Perez-Vizcaino, Duarte, Jimenez, Santos-Buelga, & Osuna, 2009; Schroeter et al., 2006; Williamson & Manach, 2005). In consequence, this is significantly affected by various factors such as sun exposure, climate, type of soil, among others (Bohn, 2014). The colonic microflora metabolizes most of the polyphenols before they get absorbed, resulting in products partly accounting for their systematic properties (Landete, 2012). The antioxidant properties of polyphenols are responsible for their antiapoptotic actions, vasodilating, antilipidemic, antithrombotic, anti-inflammatory, and antiatherogenic properties (Actis-Goretta, Ottaviani, & Fraga, 2006; Pal et al., 2003; Quinones, Miguel, & Aleixandre, 2012; Zern et al., 2005). Importantly, the antioxidant capacity of polyphenols is a 100-fold higher than that of carotenoids or vitamin E and 10-fold higher than that of vitamin A (Okoth, Chenia, & Koorbanally, 2013). The molecular mechanism associated with the antiatherogenic and antilipidemic properties of polyphenols stems from their ability to regulate the expression of various genes linked with

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energy metabolism and immune system, and/or through the histone acetylation (Ruiz, Braune, Ho¨lzlwimmer, Quintanilla-Fend, & Haller, 2007), epigenetic regulation (Berghe, 2012), expression modulation of some microRNAs (Blade´, Baselga-Escudero, Salvado´, & Arola-Arnal, 2013), and induction of changes in the methylation pattern of DNA CpG islands (Fang et al., 2003; Kato et al., 2008). In this regard, the significant role of polyphenols in the prevention of hyperlipidemia in the mice has been driven via regulating the expression of hepatic microRNAs: miR122 and miR103/107 (Joven et al., 2012). Likewise, quercetin has been reported to inhibit the histone acetyltransferase activity in the gene’s promoter region linked with the manifestation of inflammation (Ruiz et al., 2007). The authors revealed that genistein use can elicit endothelium-dependent vasorelaxation in vivo (Walker et al., 2001) and in vitro (Figtree et al., 2000). The isoflavone dihydrodaidzein has also been found to improve the endothelial function (Shen, White, Husband, Hambly, & Bao, 2006). The flavonoids have been reported to significantly control atherosclerosis owing to their hypochlorite scavenger activity (Firuzi, Mladenka, Petrucci, Marrosu, & Saso, 2004). Resveratrol, biochanin A, daidzein, and epigallocatechin gallate as part of the polyphenols group have been reported to hold significant anticancer properties (Aggarwal et al., 2004; Chung et al., 2005; Qian, Wei, Zhang, & Yang, 2005). The aforementioned conclusions demonstrate that polyphenols from plant sources have indeed emerged as an area of enormous interest in defining innovative strategies of disease control and prevention.

11.3 Roselle uses in traditional medicine Roselle is consumed as a beverage in West Africa, United States, India, Iran, Mexico, Thailand, Taiwan, Egypt, among others. The various parts of the Roselle plant have been used in folk medicine for the treatment of urinary tract infections, toothaches, hangovers, and cold (Maganha et al., 2010). In Africa, India, and Mexico, the ethnomedicinal studies have revealed the use of Roselle calyces or leaves infusion for the treatment of hypotensive, choleretic and diuretic effects, hyperlipidemia, stimulating intestinal peristalsis, and to decrease the viscosity of blood (Riaz & Chopra, 2018). In Thai traditional medicine, it is used to treat the urinary bladder and kidney stones (Maganha et al., 2010). The dried powder and extract of calyces supplemented with common salt is used to cure diarrhea, flatulence, waist pain, dysentery, and other gynecological disorders in postdelivery cases of humans and animals (Singh, Sureja, & Singh, 2006). The calyx infusion, also known as Sudan tea, is a useful remedy for biliousness, coughing, and body temperature (Padmaja, Sruthi, & Vangalapati, 2014). In Egypt, calyx is used to treat nerve and cardiac diseases and to enhance urine production. While the infusion of calyces in Guatemala is used in the treatment of drunkenness, the same is

11.4 Medicinal and therapeutic health benefits of Roselle

used to treat coughs, sore throats, and genital problems in North Africa; however the leaves are used in the treatment of abscesses and external wounds (Neuwinger, 2000).

11.4 Medicinal and therapeutic health benefits of Roselle 11.4.1 Antihypertensive activity Hypertension is a chronic medical condition with elevated arterial blood pressure. Water and sodium retention plays an important role in the progression of hypertension. Extracts from different parts of the Roselle have shown efficacy in lowering blood pressure in hypertensive humans (Inuwa et al., 2012). Hibiscus plant serves as a hypotensive agent and first line of defense against an increased level of hypertension (Walton, Whitten, & Hawrelak, 2016). The crude extract of the Roselle has the great ability to activate nitric oxide synthase enzyme and the nitric oxide pathway in the presence of endothelium. It has a weak vasorelaxation effect in the absence of endothelium, leading to the belief that the crude extract of hibiscus may have endothelium-dependent mechanisms. Another vasorelaxation mechanism induced by anthocyanin is via potassium channel activation. The relaxations obtained with the crude extract in vessels without endothelium, even if they are significantly lower compared to those observed in vessels with an intact endothelium, have led to the assumption that there is a direct relaxing effect of the extract on vascular muscles. The likely mechanism is hyperpolarization of the membrane after direct activation of potassium channels. The butanoic extract having the biggest proportion of anthocyanins carries the largest vasorelaxant activity (Al, Anwar, & Eid, 2015). Besides, the Roselle extract inhibits angiotensin-converting enzyme activity, has diuretic properties, affects endothelium-derived nitric oxide
cGMP relaxant pathway, and inhibits calcium (Ca21) influx into vascular smooth muscle cells. It also acts as a cholinergic and reduces the diffusion distance between capillaries and myocytes. Recently, the antihypertension effect of Roselle has been shown in diabetic patients and postpartum mothers (Liu, Liang, Li, & Yu, 2020).

11.4.2 Anti-inflammatory activity Inflammation is a physiological reaction that occurs in the response to different injuries or disease-causing agents (physical trauma or bacterial infection), thus limiting the damage caused by pathogens and promote tissue repair (Patil & Munoli, 2017). Although inflammation provides a protective shield from cell injury caused by genetic defects, severe trauma, chemical, and physical agents, if remains unchecked, it can lead to pathogenesis of chronic inflammatory diseases such as obesity, atherosclerosis, insulin resistance, bowel diseases, multiple sclerosis, and arthritis (Hanauer, 2006). There are generally two types of

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inflammation, acute and chronic. Acute inflammation is characterized by quick onset for a very short period and is marked by a loss of plasma protein and fluids from the blood vessels (Murakami & Hirano, 2012). Contrarily, chronic inflammation starts within 2
4 days of the initial response and it can last for months or years. It is manifested by the presence of macrophages and lymphocytes, leading to tissue necrosis and fibrosis. Anthocyanin extracted from the dried calyx of Roselle has been reported to play a vital role in anti-inflammatory activity. Different compounds such as aglycons, cyaniding, and delphinidin may suppress the expression of inflammatory mediators, that is, prostaglandin and cyclooxygenase (Hou, Yanagita, Uto, Masuzaki, & Fujii, 2005). During chronic inflammation, the primary cells released in the form of macrophages produce a huge amount of nitric oxide mediators along with proinflammatory cytokines. Recent studies indicate that anthocyanins obtained from Roselle mainly delphinidin-3-sambubioside and delphinidin are effective in suppressing the expression of nitric oxide mediators, that is, inducible nitric oxide synthase (iNOS), nitric oxide (NO), interleukin 6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), along with cytokines such as tumor necrosis factor alpha (TNF-α) and MCP-1 (Sogo et al., 2015). The in vitro antiinflammatory activity of Roselle leaves extract has also been evaluated against the RAW 264.7 murine macrophage cells by measuring the inhibition of nitrous oxide synthesized (Zhen et al., 2016). The outcome of the findings explicated dosedependent inhibition of nitric oxide synthase with wider variation among different accessions of the plant. This anti-inflammatory response of Roselle extract was attributed to quercetin, kaempferol, and chlorogenic acid. During inflammation, lysosomal enzymes are released which lead to multiple diseases. The activity of these extracellular enzymes is controlled by the magnitude of acute and chronic inflammation. Sometimes, the nonsteroidal drugs used during inflammation act by stabilizing the cell membranes of these enzymes, thereby inhibiting their activity. It is pertinent to mention that the membranes of human erythrocytes are similar to these lysosomal enzymes. The cell membrane stabilization property has been demonstrated through in vitro experiments using methanolic extract of Roselle flowers (Garbi et al., 2017). Roselle extract has also been found effective in the inhibition of protein (albumin) denaturation. The inflammation results in protein denaturation leading to the loss of its tertiary structure (Chandra, Chatterjee, Dey, & Bhattacharya, 2012). Another study has demonstrated both in vitro and in vivo anti-inflammatory properties of polyphenols extracted from Roselle. The in vitro anti-inflammatory assay was carried out on nitrite and prostaglandin E2 in lipopolysaccharide (LPS)treated RAW 264.7 cells, whereas in vivo examination was carried out on LPSinduced hepatic inflammation in rats. The results exhibited a 94.6% reduction in xanthine oxidase in vitro activity with a simultaneous decrease in nitrite and prostaglandin E2 secretions in cultured cells after the application of polyphenols (Kao et al., 2009). Likewise, in animal models, polyphenols administration significantly reduced the serum alanine and aspartate aminotransferase levels. Moreover, liver

11.4 Medicinal and therapeutic health benefits of Roselle

lipid peroxidation and lesions exhibited a noticeable decrease, whereas catalase and glutathione activity was increased. The authors explained that this antiinflammatory effect was possibly due to the downregulation of cyclooxygenase-2, p-c-Jun N-terminal kinase, and p-P38 (Kao et al., 2009). The anti-inflammatory properties of methanolic extract of Roselle seeds have been studied in rat models. Purposely, the edema was introduced in the right hind paw of rats by injecting 1% suspension of carrageenan (0.1 mL). The treatment groups were administered with Roselle extract at the doses of 200 and 400 mg/kg of weight of the body, respectively. The paw thickness of each rat was calculated using Zeitlin’s apparatus before carrageenan injection and every hour up to 6 hours after carrageenan injections. The results exhibited a significant decrease in proximal paw edema in a dose-dependent manner, suggesting the antiinflammatory activity of Roselle seeds (Ramadevi, Rao, & Rajeshwari, 2016). The seeds of the plant contain significant quantities of sterols, saponins, and flavonoids that are responsible for anti-inflammatory properties.

11.4.3 Antiobesity activity Increased body weight and fat mass lead to metabolic disorders such as obesity. In developing countries, it is one of the major public health problems and is associated with an increased incidence of other chronic diseases such as type-2 diabetes, high blood pressure, heart disease, and certain types of cancer (Segal, Rayburn, & Mart´ın, 2016). Diet, genetic disorders, sedentary lifestyle, and psychological factors are the main risk factors for obesity. Besides, high-fat diet and an imbalance between energy input and output are the major drivers for the onset of obesity, leading to excess fat accumulation in adipose tissue (Lee, Lee, Jang, Kwon, & Nam, 2017). To combat the menace of obesity, several approaches have been adopted including clinical, lifestyle changes, exercise, dietary management, and pharmacological interventions via weight loss drug. However, these approaches have certain limitations and are not sustainable (Annamalai, Mohanam, Alwin, & Prabhu, 2016). Therefore the use of plant bioactive compounds has emerged as a novel therapy, gaining momentum in current research to fight against obesity and other related metabolic disorders. Previous studies evidence the use of flavonoids and phenols in obesity treatment. For example, anthocyanins from Roselle are potent therapeutics against obesity (Ali et al., 2005; Sa´yago-Ayerdi, Arranz, Serrano, & Gon˜i, 2007). Many natural compounds in Roselle have shown promise in weight loss, fat accumulation, and diet-induced obesity. Purposely, the plant and its products have been widely consumed to treat abdominal obesity and overweight (Ganesan, Ramasamy, & Gani, 2013). Polyphenols from Roselle are responsible to counteract obesity and its related complications by modulating different key molecular targets at the cell, as well as through epigenetic modifications. The multitargeted mechanism includes the regulation of energy metabolism, transcription factors, hormones, and digestive enzymes, etc. (Herranz-Lo´pez et al., 2017).

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Another strategy to treat obesity is to inhibit pancreatic lipase, which consequently will decrease lipid absorption in the intestine. The underlying concept behind this phenomenon is to decrease the concentration of fat metabolizing enzymes. Because when any dietary fat enters the intestine, it is absorbed there and undergoes catabolism by the action of intestinal enzyme namely pancreatic lipase. Therefore lipase activity inhibition is widely considered as one of the most important indicators of an antiobesity potential of natural products (Marrelli, Loizzo, Nicoletti, Menichini, & Conforti, 2013). Consumptions of Roselle extract may reduce body weight, lipid accumulation, and total cholesterol metabolism, in both animals and humans. It is effective in inhibiting pancreatic lipase and adipocyte differentiation, thus confirming its potential in obesity management. The regular use of extract as an active ingredient of diet at a normal dosage is considered safe (Ojulari, Lee, & Nam, 2019). Modulation of signaling pathways, energy metabolism, regulation of redox homeostasis and inflammation, restoration of mitochondrial function, and regulation of epigenetic machinery are key processes involved in the molecular effects of Roselle polyphenols on obesity.

11.4.4 Antidiabetic activity All parts of Roselle such as leaves, seeds, fruits, and roots are used in many foods, nonpharmacological drugs, and herbal medicines (Riaz & Chopra, 2018). Roselle extract has numerous applications in the management of diseases such as diabetes, high blood pressure, fever, liver diseases, and so on. Besides, its therapeutic applications as an anti-inflammatory, antihyperlipidemic, and antihypertensive agent are also well documented. However, efficacy studies of Roselle against diabetes are relatively few when compared to its other applications, that is, antihypertensive or antihyperlipidemic (Bule, Albelbeisi, Nikfar, Amini, & Abdollahi, 2020). Diabetes is the most common chronic disease caused by the inadequate production of insulin. A recent study reported that the Roselle extract helped in the regeneration of pancreatic beta-cells (Adeyemi & Adewole, 2019). Notably, the flavonoid-rich aqueous extract of Roselle calyx has antidiabetic potential. Purposely, methanolic extract of calyx was used to examine its antidiabetic effect against type-1 diabetes in Wistar rats induced through 80 mg/kg body weight streptozotocin (STZ) injection. The test group of rats received 1.75 g/kg/body weight of calyx extract for 15 days. Stereological analysis demonstrated that calyx extract treatment unusually increased the density of beta-cells and improved the volume of pancreatic islets that is depleted by the STZ injection. The authors concluded that the antioxidant potential of Roselle calyx extracts induced betacells regeneration in pancreatic islets and resulted in antidiabetic properties. Likewise, another study was conducted to characterize methanolic extract of Roselle leaves and evaluate its hypoglycaemic and hypolipidemic properties in alloxan-induced diabetic Wistar rats (Ndarubu et al., 2019). The rats were administered a variable oral dose of 100, 200, and 400 mg/kg body weight for 21 days. The results explicated that Roselle leaves methanolic extract contained alkaloids,

11.4 Medicinal and therapeutic health benefits of Roselle

tannins, flavonoids, and phenols in different concentrations. Regarding efficacy, the administration of leaf extracts demonstrated dose-dependent antidiabetic activity. Furthermore, the extracts also led to a significant reduction in serum cholesterol, low-density lipoprotein cholesterol (LDL-C), and triglycerides while increasing the high-density lipoprotein (HDL) level when compared to the control group. Evidently, the Roselle leaves extract was effective in inhibiting dyslipidemia and hyperglycemia in diabetic rats. The antidiabetic property of Roselle extracts was evaluated in type II diabetic rats (Singh, Khan, & Hailemariam, 2017). The rats were fed with high-fat diet. The results reported that Roselle extract administered @ 200 mg/kg worked best and demonstrated antiinsulin resistance characteristics, thereby significantly reducing hyperinsulinemia and hyperglycemia. Furthermore, the extract was effective in lowering serum cholesterol levels. The authors further reported that Hibiscus acid [hibiscus type (2S,3R)-hydroxycitric acid lactone] is a potent inhibitor of intestinal α-glucosidase and pancreatic α-amylase activity. It is worth mentioning that pancreatic α-amylase and intestinal α-glucosidase help in complex carbohydrate digestion and convert them into simpler ones (i.e., monosaccharides) thereby, playing a significant role in postprandial hyperglycemia. Thus Roselle extract is an effective inhibitor of pancreatic α-amylase and helps in glycemic regulation. An in vitro assessment of the Roselle methanolic extract, subsequently partitioned into various fractions through different solvents varying in polarity (n-hexane, ethyl acetate, butanol), was carried out (Shadhan & Bohari, 2017). The in vitro assays included inhibitory action on glucose diffusion, α-glucosidase enzyme, and wound healing capacity. The results revealed that 50 mg/mL concentration of ethyl acetate fraction resulted in significant inhibition of α-glucosidase activity. It was further noticed that the same concentrations of methanolic fruit extract and other solvent fractions were not as potent in α-glucosidase inhibition. Likewise, ethyl acetate and butanol fractions of the methanolic extract exhibited significantly higher glucose diffusion inhibitory activity. Therefore, the methanolic extract of the fruit and its fractions can inhibit the glucose diffusion and α-glucosidase enzyme and may be useful in the glucose regulation of diabetics. Another in vitro study has explored the inhibitory capacity of Roselle infusion against α-amylase and α-glucosidase (Pe´rez-Ram´ırez, Castan˜o-Tostado, Ram´ırez-de Leo´n, Rocha-Guzma´n, & Reynoso-Camacho, 2015). The maximum inhibition reached was 37% and 25% for α-amylase and α-glucosidase, respectively with the dose of 30 mg dried Roselle per mL. The authors attributed this inhibition of the digestive enzymes to the phenolic compound, hibiscus acid, and cyanindin-3-glucoside whose hydroxyl groups may lead to the formation of hydrogen bonds with the polar groups of the allosteric site of the enzymes, thus changing the molecular configuration and hydrophobic or hydrophilic properties of the enzymes thereby decreasing their activity. This signifies the usage of Roselle beverages in the form of infusions, tea, and other drinks to derive associated health benefits.

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However, the efficacy may vary depending upon parameters of extraction, processing conditions, the concentration of bioactive compounds, storage, and the presence of other ingredients.

11.4.5 Nephroprotective activity Diabetic nephropathy is associated with type-2 diabetes which may ultimately lead toward end-stage renal disease. Nephropathy is considered among the major microvascular disorders related to diabetes. The renal lesions produced during type-1 and type-2 diabetes are also comparable. Increased kidney size, glomerular volume, and urinary albumin excretion are some of the indicators of nephropathy. Last-stage diabetic nephropathy is characterized by progressive renal insufficiency, hypertension, and proteinuria which consequently proceeds toward kidney tissue damage (Das, Vasudeva, & Sharma, 2019). Nephroprotective role of Roselle has been demonstrated in various studies. Recently, nephroprotective properties of aqueous extract of Roselle were evaluated in the animal model (Okonkwo et al., 2020). Purposely, male adult albino rats were treated with 50 mg/kg body weight lead acetate for 28 days to induce degeneration of kidney tissues, causing the elevation in blood urea and creatinine levels which are key biomarkers of kidney damage. Subsequently, the rats were fed with 500 mg/kg body weight aqueous extract of Roselle. The outcomes of the intervention demonstrated a significant decrease in blood urea and creatinine levels, indicating restoration of kidney architecture. However, a nonsignificant change in serum electrolytes was observed. Another intervention study was conducted to evaluate the nephroprotective role of Roselle (Ibrahim & Noman Albadani, 2014). To induce nephrotoxicity, experimental rabbits were given gentamicin (80 mg/kg body weight) for a week followed by administration of Roselle extract at a dose of 250 mg/kg body weight. A significant decrease in nonenzymatic markers of kidney dysfunction was observed through in vivo studies. Oxidative stress is considered as one of the contributing factors in diabetic nephropathy. STZ-induced diabetic rats were fed with polyphenol extract from Roselle, purportedly effective in treating nephropathy. Significant reduction in kidney mass was witnessed after extract administration when compared to the control group (Lee et al., 2009). It is pertinent to mention that STZ injection leads to increased kidney biomass. It is believed that oxidative stress may be a major contributor to diabetic nephropathy. Therefore the antioxidant potential of Roselle plays a significant role in the management of diabetic nephropathy. An experiment was conducted to evaluate the role of Roselle aqueous extract against diabetic nephropathy in STZ-induced type 1 diabetic rats (Wang et al., 2011). Evidently, the aqueous extract of the Roselle was able to significantly reduce the lipid peroxidation while increasing the glutathione and catalase activities in diabetic kidneys. Moreover, the histological examination of diabetic rats revealed that extract treatment resulted in improved hyperglycemia-caused osmotic diuresis in renal proximal convoluted tubules.

11.4 Medicinal and therapeutic health benefits of Roselle

Another study was conducted to evaluate the efficacy of Roselle extract against Adriamycin (10 mg/kg) induced nephrotoxicity in male rats. The authors noticed a significant improvement in kidney functions after treatment with Roselle extract. Moreover, it led to the normalization of malondialdehyde and glutathione peroxidase levels. Likewise, the effects of Roselle supplementation on renal biomarkers and various electrolytes were evaluated in obese Wistar male rats. After treatment with Roselle reported a significant reduction in urea, creatinine, and triglycerides occurred indicating that nephropathy associated with obesity decreased by supplementing Roselle. This protection was associated with improvement in lipid profile and correction of glomerular filtration rate (Melchert al., 2016). Besides, the Roselle has been investigated for its diuretic activity in a rat model (Alarco´n-Alonso et al., 2012). The natriuretic and diuretic effects of the aqueous extracts displayed dose-dependent behavior. There was a significant increase (48%) in renal filtration with the administration of Roselle aqueous extract. The quercetin in the aqueous extract was believed to affect vascular endothelium causing oxide nitric release that led to increased renal vasorelaxation, resultantly improving kidney filtration. Aqueous infusion of Roselle has also been investigated for its diuretic effect in rat models. The results reported that the treatment group had significantly increased urinary output when compared to the water placebo and hydrochlorothiazide group. Furthermore, urinary sodium, potassium, and uric acid also witnessed a noticeable increase (Riaz & Chopra, 2018). The uricosuric effect of Roselle has been evaluated in human subjects who were provided with the tea prepared from dried Roselle calyces @1.5 g/day twice a day for 15 days (Prasongwatana, Woottisin, Sriboonlue, & Kukongviriyapan, 2008). One group of subjects was having a history of renal stones, whereas others had no renal disease. Urine and blood samples were taken after the administration of tea for 15 days. An increase in citrate and oxalate was observed, while uric acid excretion and clearance were significantly enhanced in both groups, indicating that tea can be used for the treatment of hyperuricemia. However, the constituent involved in this uricosuric effect needs to be sufficiently characterized. Likewise, another study evaluated the effect of Roselle against oxonic acidinduced hyperuricemia in rats. Male Sprague
Dawley rats were administered intraperitoneally with oxonate solution and normal saline for 1 week. Urate enzymes (urease and xanthine oxidase) and uric acid were affected by Roselle extract treatment. The results demonstrated inhibition of oxonic acid-induced hyperuricemia, which led to a significant decrease in uric acid. These studies provide a sufficient scientific basis to infer that Roselle extract may be a useful remedy for the treatment of hyperuricemia (Kuo et al., 2012).

11.4.6 Hepatoprotective activity The liver is the most important and indispensable organ of the human body having multifunctional aptitudes. The major role of the liver is to help in the

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metabolism of carbohydrates, fats, and protein and also to remove the harmful metabolites and drug toxins. After passing through the intestinal tract, toxins directly reached to their foremost destination called the liver, where the breakdown and elimination of toxins such as drug toxins and foreign toxins occur (Abhilash, Maheswari, Gopal, & Chanda, 2014). The liver also helps in the storage of many important nutrients such as iron, glycogen, fat-soluble vitamins, and minerals. Whenever there is a decline in the status of these nutrients occur within the body, the liver ultimately balance them by releasing them from stores. It also detoxifies the chemicals, drugs, microorganism infections, heavy metals, and other foreign chemicals. The liver synthesizes important proteins such as hormones, enzymes, immune factors, and blood clotting factors (Akinloye & Olaniyi, 2011; Nayak, Jain, & Sahoo, 2011). Liver diseases are considered as one of the major health problems as it affects about 10% of the world population and the increasing global burden of diseases. The factors that cause liver damage are toxins (thioacetamide), malnutrition, over drug intake (paracetamol, chemotherapy agent CCl4), viruses, excessive alcohol consumption, among others, which may lead to liver diseases such as chronic hepatitis, fatty liver, fibrosis, carcinoma, and drug-induced injuries (Ali & Kumar, 2015; Luk et al., 2007). Previously, scientists and researchers are paying much attention to find the cure for liver disease using folk medicine. Roselle is one of the folk herbs having a hepatoprotective effect as reported previously (Tsai, Huang, Chang, & Wang, 2014). The plant extract decreases the accumulation of fatty tissues and maintains the glycemic index. Moreover, hepatic steatosis is attenuated through sterol regulatory protein 1C, interleukin-1 messenger RNA blockade and by peroxisome proliferator-activated receptor, lipid peroxidation, tumor necrosis factor-alpha and through elevating the catalyze messenger RNA (Kshirsagar, Mohite, Aggrawal, & Suralkar, 2011; Okaiyeto, Nwodo, Mabinya, & Okoh, 2018; Torres Gonza´lez et al., 2014). Previously, hepatoprotective activity of Roselle aqueous extract was analyzed in albino rats having alcohol induce liver injury. Orally about 200
400 mg/kg of aqueous extracts of Roselle leaves were given to the animal. The results reported a decrease in alanine aminotransferase, alkaline phosphatase, aspartate transaminase enzymes, and bilirubin. The hepatoprotective effect of Roselle was further supported through histopathological examination and it was observed that its extract reverses the shape of the liver disturbed through the liver intoxication. The author revealed that hepatoprotective effect of the Roselle may be due to the presence of the bioflavonoids (Bhavana, Rajani, Babu, & Bonthagarala, 2017). Roselle methanolic extracts were utilized to investigate the hepatoprotective activity against carbon tetrachloride-induced liver injury in rats. Carbon tetrachloride injection results in the elevation of alanine aminotransferase, alkaline phosphatase, aspartate transaminase enzyme, thiobarbituric acid reactive substances, and lactate dehydrogenase, besides reduction within the superoxide dismutase, reduced glutathione and catalyze. Different doses of Roselle extract ranged from 50 to 100 mg/kg were given to rats significantly reduced the liver enzymes and increased the level of

11.4 Medicinal and therapeutic health benefits of Roselle

superoxide dismutase and catalase. The author reported that phenolics extract of the Roselle is responsible for the hepatoprotective activity as the extract has the ability to absorb and neutralize the free radicals (Owoade & Adetutu, 2015).

11.4.7 Cardioprotective activity Calyces from Roselle have long been used to treat cardiac and nerve disease (Pe´rez-Torres, Ruiz-Ram´ırez, Ban˜os, & El-Hafidi, 2013). Cardiovascular effects of Roselle have long been demonstrated in different studies. Phenolics and anthocyanins such as delphinidin-3-sambubioside and cyanidin-3-sambubioside are present in significant amounts in Roselle calyces, which act as strong hydrophilic antioxidants. Likewise, protocatechuic acid, caffeic acid, catechins, and epigallocatechin gallate are the polyphenolics with proven benefits in the management of cardiovascular diseases, particularly atherosclerosis (Huang, Chang, Kao, & Lin, 2015). Because of antioxidant potential and abundance of polyphenols, Roselle extract has gained considerable attention of researchers to treat cardiovascular diseases. Arrhythmic heart-beat and other associated risk factors such as atherosclerosis can be treated with Roselle extract. The extract is beneficial in preventing oxidative damage to heart tissues. Purposely, scientists evaluated the cardioprotective impact of Roselle polyphenols against Langendorff-perfused rat hearts in a dosedependent manner. Resultantly, the Roselle polyphenols decreased systolic function of the heart as witnessed by lowered left ventricular developed pressure and dP/dt max, indicative of a negative inotropic effect. Moreover, the polyphenols administration led to negative chronotropic action with a simultaneous increase in the maximal velocity of relaxation (Lim, Budin, Othman, Latip, & Zainalabidin, 2017). However, polyphenols perfusion enhanced coronary pressure, indicating improved coronary blood flow. The aqueous infusion of the Roselle calyces contains anthocyanins, phenolic acids, and other bioactive compounds responsible for reducing the blood pressure and imparting cardioprotective functions. Roselle petals also contain numerous bioactive compounds including flavonoids and anthocyanins, indicative of its cardioprotective and inflammatory activities (Obouayeba et al., 2015).

11.4.7.1 Antihyperlipidemic The hypolipidemic effects of Roselle extract have been previously explicated in various scientific studies. High-fat diet and energy imbalance may lead to metabolic disorders such as hyperlipidemia, hypertension, and other cardiovascular malfunctions (Huang et al., 2015). Roselle has demonstrated a multitargeted mechanism that includes regulation of energy metabolism, oxidative stress, inflammatory pathways, transcription factors, hormones, peptides, digestive enzymes, as well as epigenetic modifications (Aziz, Wong, & Chong, 2013; Riaz & Chopra, 2018). The short-term consumption of Roselle is quite beneficial with infrequent harmful effects (Aziz et al., 2013). The extracts of Roselle leaves and

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calyces have explicated significant antihyperlipidemic in hyperlipidemic rats, which was comparable to standard drugs used in such treatments (Ochani & D’Mello, 2009). With the increase in HDL levels in serum, excess cholesterol passes through catabolism process through the reverse cholesterol transport pathway and is shifted from the peripheral cells to the liver, thus inhibiting the absorption of excess cholesterol. This is accompanied by the interference in lipoprotein production coupled with increased expression of LDL receptors and their protection, accelerating the removal of LDL-C. Consequently, it results in significantly higher catabolism and degradation of cholesterol in the body. All such mechanisms are involved either individually or collectively in lowering LDL-C and total cholesterol with the administration of Roselle extracts (Aziz et al., 2013). There is evidence from previous studies that Roselle extract reduced hyperlipidemia. Previously, cholesterol was fed to rabbits for 10 weeks and then treated with Roselle extract (0.5% or 1%) showed that the level of total cholesterol, LDL level, and triglyceride in the serum was reduced (Chen et al., 2003). In another study, an aqueous extract of the dried calyx of Roselle was given to hypercholestrolemic rats for 6 weeks. The results reported a significant reduction in serum cholesterol, LDL level, and triglycerides (Hirunpanich et al., 2006). Likewise, another work also reported that aqueous extract of Roselle has decreased more than 50% of serum triglyceride levels in mice under a hypercaloric diet for various weeks (Ferna´ndez-Arroyo et al., 2011).

11.4.7.2 Atherosclerosis Cardiovascular diseases, the world’s leading cause of death, are caused predominantly by atherosclerosis, a chronic inflammatory condition of the blood vessels. Atherosclerosis is a smoldering, immunoinflammatory disease of medium-sized to large arteries and is mediated by blood lipids. The major players in the development of atherosclerosis are endothelial cells, leukocytes, and intimate smooth muscle cells. Superimposed thrombosis is usually held responsible for the most devastating consequences of atherosclerosis, that is, heart attack and stroke (Hansson & Hermansson, 2011). Atherosclerosis pathogenesis requires activation of the proinflammatory signaling pathways, cytokine/chemokine expression, and increased oxidative stress. Oxidative stress is an imbalance in favor of increased reactive oxygen species (ROS) production and/or decreased innate antioxidant defenses of the body (Peluso, Morabito, Urban, Ioannone, & Serafi, 2012). ROS plays a significant role in the inflammatory response, apoptosis, cell growth, and vascular tone modification as well as in the oxidation of LDL-C, which is considered more significant in atherogenesis than native LDL (Fo¨rstermann, Xia, & Li, 2017). Therefore due to the nature of the atherosclerosis pathogenesis, therapeutic drugs aimed to treat atherosclerosis remain very limited (Orekhov, Sobenin, Revin, & Bobryshev, 2015). It has been documented that the latest antiatherosclerotic “gold standard” medications such as statins can also produce antiinflammatory effects, regardless of lipid-lowering effects (Rosenson, 2004). Consequently, anti-inflammatory therapy is a good strategy for atherosclerosis

11.4 Medicinal and therapeutic health benefits of Roselle

prevention and treatment. With the advent of an era of natural therapies, the plant bioactive compounds can translate into effective novel cardiovascular therapeutics (Fang, Little, & Xu, 2018; Shen, 2015). The fleshy bright red calyces of Roselle are enriched with polyphenols, pectin, phytosterols (e.g., β-sitosterol and ergosterol), L-ascorbic, arachidic, citric, stearic, and malic acids (Gurrola-D´ıaz et al., 2010; Sa´yago-Ayerdi et al., 2007). Many epidemiological studies support the antioxidant properties of polyphenols, though these effects depend on their bioavailability and absorption. Thus other factors such as climate, soil type, crop type, and sun exposure greatly influence the phenolic profile of the plants (Barbosa, Bressan, Zulet, & Mart´ınez, 2008). Most of the polyphenols are metabolized before being absorbed by colonic microorganisms, and the metabolites resulting from this fermentation are partly responsible for their systemic effects. The antioxidant capacity of polyphenols accounts for their vasodilatory, antithrombotic, anti-inflammatory, and antiapoptotic actions, as well as their antilipidemic and antiatherogenic effects. The antioxidant activity of polyphenols is 10 times higher than vitamin A and 100 times higher than vitamin E or carotenoids. More precisely, studies suggest that LDL oxidation and HDLs may be attenuated by phenolic compounds (Landete, 2012). For example, quercetin, the active ingredient of Roselle, has been reported to inhibit the action of histone acetyltransferase in the promoter region of genes associated with inflammation (Ruiz et al., 2007). While investigating the hypoglycemic and hypolipidemic effect of polyphenolic extract of Roselle detected at least 18 different types of phenolics. Plant-derived polyphenols have been useful for the prevention of atherosclerosis (Peng et al., 2011). Scientific studies have demonstrated the antiatherogenic function activity of Roselle polyphenols along with its role in the alleviation of hypertension and hyperlipidemia (Herrera-Arellano et al., 2007).

11.4.8 Renal effects, uricosuric effect, and hyperuricemia The renal effects of Roselle have been pharmacologically demonstrated in both preclinical animal trials (Aguwa, Ndu, Nwanma, Udeogaranya, & Akwara, 2004; Laikangbam & Devi, 2012) and clinical experiments (Herrera-Arellano, FloresRomero, Chavez-Soto, & Tortoriello, 2004; Prasongwatana et al., 2008). For instance, a study was designed to explore the diuretic effects of Roselle infusion in conscious rats. The results reported a significant increase in urine collection (103 mL/kg) when compared with hydrochlorothiazide treated group (25 mg/kg), and the water placebo (46 mL/kg). Likewise, uric acid, urinary potassium, and sodium were also increased in the treatment group (Ribeiro et al., 1988). The antilithiatic effect of Roselle extract was observed in Wistar rats fed with 3.5 mg extract on a daily basis. This led to increased oxalate excretion in urine, while calcium deposition in kidneys was decreased (Woottisin et al., 2011). Likewise, another study reported the successful application of Roselle extract in the prevention of urolithiasis in albino rats (Laikangbam & Devi, 2012). The efficacy of

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Roselle extract was investigated in oxonic acid-induced hyperuricemia rats. The acquired results revealed that Roselle extract inhibited hyperuricemia and lowered the uric acid compared to allopurinol treatment (Kuo et al., 2012). Another study reported the antihyperuricemia effect of Roselle calyx in male Wistar rats (Wahyuningsih, Sukandar, & Sukrasnoa, 2016). The effect of Roselle aqueous extract was investigated on renal Ca (2 1)-Mg (2 1)-ATPase, and Na (1)-K (1)-ATPase activities in Wistar rats. The results exhibited improved renal functioning coupled with a reduction in creatinine and serum urea concentrations. A significant decrease in renal Ca (2 1)-Mg (2 1)ATPase activity was noticed, while renal calcium remained unchanged. However, no effect on renal Na (1)-K (1)-ATPase activity was reported by either dose of the extract. The authors concluded that oral administration of Roselle extract could preserve the renal function in spite of decreased renal Ca (2 1)-Mg (2 1)ATPase activity (Olatunji, Usman, Adebayo, & Olatunji, 2012). The preclinical trial has been performed to explore the diuretic activity of Roselle aqueous extract through in vivo models. The dose-dependent behavior was observed for natriuretic and diuretic effects with a potassium-sparing effect. Moreover, the renal filtration was increased up to 48% with the consumption of Roselle aqueous extract, while the additive effect when witnessed was perfused with furosemide (Alarco´n-Alonso et al., 2012). The clinical trial accompanied by chemical characterization of Roselle aqueous extract reported natriuretic effects in patients with mild to moderate hypertension (Herrera-Arellano et al., 2004). An intervention was performed in Thailand to explore the effect of Roselle tea on 18 subjects with or without a history of renal stones. Contrary to previous evidence, the intake of Roselle tea for 15 days @ 3 g per day dosage did not demonstrate diuretic and antilithiatic effects. Nonsignificant differences were also observed in urinary volume and serum sodium, while the significant uricosuric effect was evident in normal and renal stone-forming subjects (Prasongwatana et al., 2008). A similar uricosuric effect was observed in rats administered with a decoction of Roselle calyces at an oral dose of 1 g/kg body weight (Mojiminiyi, Adegunloye, Egbeniyi, & Okolo, 2000).

11.4.9 Treatment of anemia Anemia is an iron deficiency disease characterized by low red blood cells in the blood. The red blood cells carry and deliver oxygen to the body. The Roselle is rich in ascorbic acid and iron has been one of the most cited species against anemia. The absorption of nonheme iron in anemic patients is accelerated by ascorbic acid, which explains its potential role as antianemic in traditional medicine (Peter, Rumisha, Mashoto, & Malebo, 2014). Purposely, the aqueous Roselle extract (200
1000 mg/kg body weight) was studied for hematological parameters such as hematocrit, hemoglobin along with total and differential white blood cells in rats to explore its practical applications in the treatment of anemia. The results reported a significant elevation in hemoglobin and hematocrit in the treatment

11.4 Medicinal and therapeutic health benefits of Roselle

group fed with 200
400 mg/kg body weight. However, the beneficial effects were not sustained at higher doses (Adigun, Ogundipe, Anetor, & Odetunde, 2006). Likewise, another study on Wistar albino rats reported the usefulness of Roselle extract administration for the hematopoietic system (Emelike & Dapper, 2013). The Roselle decoctions have also been used as a source of iron for the treatment of anemia. The results explicated that dry fermented Roselle calyces had lower pH, which increases mineral bioavailability. Moreover, the higher concentration of ascorbic acid plays an important role in enhancing the mineral bioavailability (Falade et al., 2005). Clinical intervention with aqueous Roselle extract and spanning over 30 days in mild to moderate anemic subjects did not enhance the iron status in malaria-endemic region. The results reported a nonsignificant increase in serum ferritin in the test group but the increase in the control group was significant. Likewise, the C-reactive protein and hemoglobin showed no variation in either of the control or test group (Peter, Rumisha, Mashoto, Minzi, & Mfinanga, 2017). In another study, the effect of Roselle flower extract was investigated in pregnant anemia women taking iron supplements. The results revealed that the administration of extract accompanied by iron tablets significantly enhanced the hemoglobin levels, possibly due to higher ascorbic acid present in Roselle (Soejoenoes & Wahyuni, 2017). Another study was designed with a different preparation of Roselle drinks to compare and investigate hematological parameters. The results reported a significant increase in hemoglobin, platelet count, packed cell volume, and red blood cell count, whereas lymphocytes and white blood cells were significantly decreased in Wistar albino rats. The study concluded that Roselle drinks exhibit hematocrit properties and may be used for the treatment of anemic patients. Nevertheless, more studies are needed to elucidate the mechanism and long-term effects before recommendations could be made (Chukwu, Ikewuchi, & Akaninwor, 2018).

11.4.10 Cancer preventive activity Roselle is enriched with phenolic compounds, including protocatechuic acid. Protocatechuic acid has demonstrated in vitro protective effects against tert-butylhydroperoxide induced genotoxicity and cytotoxicity in hepatocytes through radical quenching and DNA repair (Tseng et al., 1998). It has also inhibited 12O-tetradecanolyphorbol-13-acetate induced skin tumor formation in CD1-mice (Tseng et al., 1998) and suppressed the survival of human promyelocytic leukemia in HL-60 cells via the reduction of Bcl-2 expression and retinoblastoma phosphorylation (Tseng et al., 2000). Similar effects in human gastric carcinoma were also reported where apoptosis could be mediated by p38 MAPK/FasL cascade pathway and via p53 phosphorylation (Lin, Huang, Huang, Chen, & Wang, 2005). The anthocyanins in Roselle such as delphinidin-3-sambubioside promote apoptosis against human leukemia and smooth muscle cells via the Bid pathway,

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p38-FasL pathway (Chang et al., 2005; Hou, Tong, Terahara, Luo, & Fujii, 2005), p53 pathway, and p38 pathway (Lo, Huang, Lin, Chien, & Wang, 2007). The chemopreventive role of Roselle extract on 2-amino-1-methyl-6-phenylimidazo [4,5-b] pyridine and azoxymethane-induced aberrant crypt foci formation in F344 rats has also been studied. The results reported a significant reduction in aberrant crypt foci formation, thus demonstrating antimutagenic activity of Roselle extract in the initiation stage, though in postinitiation stage the number of aberrant crypt foci was increased (Chewonarin et al., 1999). In another study, Roselle leaf extract was explored against prostate cancer through in vivo and in vitro experiments. The results reported antiapoptotic effect mediated by both extrinsic (Fas-mediated caspase 8/t-Bid), and intrinsic (Bax/cytochrome c-mediated caspase 9) pathways. The growth of prostate tumor xenograft was also inhibited in athymic nude mice. The authors concluded that leaves instead of calyces are a significantly larger source of polyphenolic compounds (Lin et al., 2012). The effect of Roselle anthocyanins was explored on N-nitrosomethylurea-induced leukemia in male Sprague
Dawley rats. The results reported inhibition of leukemia by 33.3% at 0.2% dose of extract. Besides, anthocyanins rich extract improved the biomarkers of histopathology, hematology, and morphology. However, the level of uric acid, alanine transaminase, myeloperoxidase, and aspartate aminotransferase was reduced (Tsai et al., 2014). Another study has also reported in vivo antileukemic activity of delphinidin-rich Roselle extracts (Wu et al., 2016). The anthocyanins rich extract modified the functions of mitochondria and stimulated cell death through necrosis and autophagy in MCF-7 cells rather than programmed cell death. The Roselle juice has also been reported to have an antiproliferative effect on cervical (HeLa), ovarian (Caov-3), and breast (MAD-MB-231 and MCF-7) cancer cells (Akim, Ling, Rahmat, & Zakaria, 2011). Likewise, the significant effect of Roselle calyces extract has been reported on the cytotoxicity of leukemia cells (K-562) (Formagio et al., 2015). The Roselle extract was investigated against fetal foreskin fibroblast and human breast adenocarcinoma cell line for cytotoxic effects. The results revealed that Roselle extract led to significantly more apoptosis in MCF-7 cells. However, the extract was not cytotoxic against normal fetal foreskin fibroblasts (Khaghani et al., 2011). The above studies explicate that phenolic compounds present in Roselle possess significant anticarcinogenic property; however, thorough in vivo studies are required to further elucidate this effect for practical applications in the food and pharmaceutical industry.

11.4.11 Uses against cadmium poisoning The epidemiological studies have provided evidence on the close connection between the quality of diet, environmental pollution, and human health (Nogawa et al., 2017; Satarug, Vesey, & Gobe, 2017). Recently, researchers have focused their attention on cadmium toxicity owing to its presence in the food and environment (Mezynska & Brzo´ska, 2018; Satarug et al., 2017). The scientific reports

11.4 Medicinal and therapeutic health benefits of Roselle

have shown that low-level exposure to cadmium generates a higher risk for human health, including damage to the liver (Walton et al. 2016; Wang et al., 2016). Therefore the World Health Organization, nutritionists, and toxicologists are increasingly interested in diagnosing effective ways to avert the major health effects of cadmium exposure (M Brzo´ska, Borowska, & Tomczyk, 2016; Pru¨ss¨ stu¨n, Wolf, Corvala´n, Bos, & Neira, 2016). U Recent scientific reports have assessed the Roselle extract against cadmiuminduced hepatotoxicity in male Wister rats. The results revealed that aqueous extract of Roselle calyces (0.2 g/kg) significantly reduced cadmium toxicity by protecting liver and testis lipoperoxidation (Omonkhua, Adesunloro, Osaloni, & Olubodun, 2009). The other study has reported the immunoprotective effect of anthocyanins-rich Roselle extract. The Roselle extract improved the viability of cadmium suppressed cells and decreased the production of cadmium-mediated macrophage-activation markers in a dose-dependent manner when compared to quercetin dihydrate (Okoko & Ere, 2012). The possible prevention against cadmium hepatotoxicity was attributed to the higher antioxidant properties of the Roselle plant (Me˛˙zy´nska & Brzo´ska, 2019). Likewise, a study was designed to explore the anthocyanins extracted from Roselle calyces against cadmiuminduced oxidative stress in Wister rats. The outcome revealed that pre- and posttreatment with anthocyanins-rich extract improved catalase, superoxide dismutase, and glutathione-s-transferase activity. The increase in glutathione level and reduction in tissue lipid peroxidation also confirmed the antioxidant properties of anthocyanin-rich Roselle extract in ameliorating cadmium-induced oxidative stress (Orororo, Asagba, Tonukari, Okandeji, & Mbanugo, 2018a, 2018b, 2018c). The antioxidant potential of Roselle flower extract was evaluated against cadmium-induced oxidative stress in the liver tissues of rats. The outcome of the study revealed that pretreatment with Roselle flower extracts significantly improved the biochemical modifications induced by cadmium. Therefore Roselle extract offers good protection against hepatotoxicity caused by cadmium and may find clinical application against different drugs and toxins which otherwise may cause damage to the cellular network through the generation of ROS (AlKubaisy, Al-Groom, & Al-Amoush, 2016). Similarly, in another study Roselle calyces extract was explored against cadmium-induced kidney and liver injuries in male Wister rats. The acquired results revealed that treatment with aqueous Roselle extract significantly increased the activities of catalase, and superoxide dismutase, while significantly lowering the malondialdehyde concentration in the kidney. Besides, aqueous extract treatment significantly reversed the effect of cadmium on the activities of alkaline phosphatase, alanine aminotransferase, and aspartate aminotransferase in the serum of rats. The concentrations of urea and creatinine were attenuated, whereas elevated protein concentration was reported in a group of rats treated with the extract. The results showed that Roselle extract was better in protection against cadmium-induced kidney and liver hepatorenal toxicity when compared to the silymarin effect (Ebhohon, Ibeh, Ejiofor, Abu, & Chibueze, 2019).

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The role of anthocyanins rich Roselle extract has been explored in the production of reproductive hormones in male Wister rats having cadmium-induced toxicity. The pretreatment with Roselle extract significantly normalized the reproductive hormones. The effect of Roselle extract was reported to be connected with the antioxidant capabilities of the anthocyanins (Orororo et al., 2018a, 2018b, 2018c). Likewise, another study has also reported that pretreatment with anthocyanins rich Roselle extract resulted in amelioration of cadmium mediated activities of catalase, superoxide dismutase, glutathione, and testicular weight in the rat. The Roselle extract administration has also reduced the lipid peroxidation that could be related to anthocyanin properties of the extract (Orororo et al., 2018a, 2018b, 2018c).

11.5 Future perspectives The available scientific evidence suggests numerous potential health applications and therapeutic uses of Roselle against various ailments. However, there is a greater need for controlled, randomized, and robust clinical trials with wellcharacterized Roselle preparation to document its usefulness in various health applications. The studies on the mechanism of action against cancer are limited and should be executed before any recommendations are made for cancer patients. Well-designed research on the antianemic activity of Roselle is also required with a bigger sample size to establish its efficacy and elaborate mode of action. The previous studies have confirmed the safety of Roselle consumption at low dosage since no adverse effects on the kidney or liver were observed (Riaz & Chopra, 2018). Therefore Roselle can be used in the management of various degenerative diseases as an active ingredient of functional foods or nutraceuticals. Further studies are recommended on the potential pharmacological applications of Roselle. The efficacy of Roselle extract for the treatment of type-2 diabetes needs further investigations as previous studies have reported promising prospects against hypertension and hyperlipidemia; conditions strongly linked with metabolic syndrome or type-2 diabetes (Al-Anbaki et al., 2019; Peng et al., 2011). The methanolic extract of Roselle leaves exhibited significant neuroprotective activity against 6-hydroxydopamine-lesioned induced toxicity in the rat model of Parkinson’s disease (Hritcu et al., 2011). However, clinical trials are needed, and mode of action should be explored before pharmacological applications. Future research efforts should be geared toward gynecological applications as Roselle roots have shown promising effects in uterotropic activity, fertility regulation, and antiimplantation (Vasudeva & Sharma, 2008). Most of the previously published scientific opinions do not establish a correlation between the chemical profile of the prepared extracts and potential therapeutic uses. Therefore better standardization of extract, chemical profiling, and correlation with therapeutic/pharmacologic applications need to be explored in future studies.

References

11.6 Conclusion The present work has summarized most of the previous studies published on traditional uses of Roselle, potential bioactive constituents, and their therapeutic applications. The previous studies provide significant evidence for the widespread use of Roselle infusion or decoction for the management of chronic and degenerative diseases such as hyperlipidemia, hypertension, diabetics, cancer, hepatoprotection, nephroprotection, among others. However, these ethnomedicinal studies provide little to no guidance about the cultivation, preparation of infusion, and consumption patterns. Also, the published work does not define any specific dose of Roselle decoction with respect to its therapeutic and medicinal properties. Besides, various extraction methods for better retention of extract efficacy need to be part of future work. Conclusively, the work provides substantial evidence related to the nutraceutical potential of Roselle for the prevention of various chronic and lifestyle-related diseases and merits further investigations.

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CHAPTER

Hibiscus sabdariffa interactions and toxicity

12

Haroon Elrasheid Tahir1, Gustav Komla Mahunu2, Zou Xiaobo1 and Abdalbasit Adam Mariod3,4 1

School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China Department of Food Science and Technology, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana 3 Indigenous Knowledge and Heritage Center, Ghibaish College of Science & Technology, Ghibaish, Sudan 4 College of Sciences and Arts-Alkamil, University of Jeddah, Alkamil, Saudi Arabia

2

Chapter Outline 12.1 Introduction ...............................................................................................187 12.2 Hibiscus
drug interactions .........................................................................189 12.3 Hibiscus sabdariffa toxicology ....................................................................191 12.4 Effect of hibiscus on kidney and liver functions ............................................193 12.5 Other adverse effects ..................................................................................194 12.6 Conclusion .................................................................................................194 References ..........................................................................................................195

12.1 Introduction Natural products have been utilized from the ancient period for drink preparation based on their potential medicine. In addition to the promising effect to reduce the risk factors associated with metabolic syndrome, its safety and toxicological characteristics should be considered. The herbal medicine contained in hibiscus is also commonly utilized as a basic component of plant-based commercial medicines, often accounting for 50% of the total constituents (Nunes, Rodrigues, Alves, & Oliveira, 2017). Several studies have demonstrated the scientific basis for the report on various parts of hibiscus tea that have been used to cure colds, urinary tract infections, hangovers as well as cardiovascular diseases (Guardiola & Mach, 2014; Riaz & Chopra, 2018). However, herbal products might not be safe for curing individual or vulnerable groups (including children and pregnant women) because of the Roselle (Hibiscus sabdariffa). DOI: https://doi.org/10.1016/B978-0-12-822100-6.00014-8 © 2021 Elsevier Inc. All rights reserved.

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herbal
synthetic drug interactions (Nunes et al., 2017). Similar to drug
drug interactions, the multicomponents of herbs can exert their impacts on pharmacokinetics and thus cause herb
drug interactions (Chan, Tan, Xin, Sudarsanam, & Johnson, 2010). The adverse effect arising from herb
drug interactions may be mild, moderate, or even lethal, dependent on many factors associated with the patients, herbs, and drugs (Yang, Hu, Duan, Zhu, & Zhou, 2006). Till date, herb
drug interactions were often unsuspected by medical doctors for many reasons. Most trained medical doctors lack adequate knowledge of herbal medicines and their possibilities for drug interactions (Rosenkranz, Fasinu, & Bouic, 2012). Herb
drug interactions take place when the pharmacokinetic profile of either product is changed substantially due to their coadministration that induces adverse effects, toxicity, or therapeutic failure (Jacquin-Porretaz et al., 2017; Johnson, Oyelola, Ari, & Juho, 2013). Previous studies have shown that the hibiscus infusion leads to a decrease in the elimination of diclofenac and hydrochlorothiazide (Fakeye, Pal, Bawankule, Yadav, & Khanuja, 2009; Hopkins, Lamm, Funk, & Ritenbaugh, 2013). On the other hand, Johnson et al. (2013) studied the in vitro inhibitory activities of the aqueous extract of hibiscus on selected cytochrome P450 Isoforms and found that this might not lead to the considerable herb
drugs interaction. Several researchers have reported the toxicity properties of phytochemicals in herbs and species (Ali, Blunden, Tanira, & Nemmar, 2008; Bode & Dong, 2011; Dasgupta & Hammett-Stabler, 2011; Zhang et al., 2004). Pearl, Drillings, and Conry (2011) reported various toxicological effects such as the central nervous system, heart, gastrointestinal system, lungs, and autonomic nervous system in the simultaneous consumption of American hellebore (Veratrum viride). Also, several toxic effects were observed when herbs/seeds used such as Blue Cohosh (Caulophyllum thalictroides) and betony (Stachys officinalis) (Connor, Davidson, & Churchill, 2001; Pearl et al., 2011). Hibiscus infusions have an ancient traditional utilization both in food and drugs and generally are reported to be risk-free (Da-CostaRocha, Bonnlaender, Sievers, Pischel, & Heinrich, 2014). The therapeutic properties and chemical composition were affected by seasonal and geographical variations (Tahir et al., 2020; Tahir, Xiaobo, Jiyong, Mariod, & Wiliam, 2016). Previous studies on animals and humans showed some adverse effects (Akindahunsi & Olaleye, 2003; Gaya, Mohammad, Suleiman, Maje, & Adekunle, 2009; Jacquin-Porretaz et al., 2017; Onyenekwe, Ajani, Ameh, & Gamaniel, 1999; Orisakwe, Husaini, & Afonne, 2004). Other studies have reported that the extract has an estrogenic effect (Ali, Salih, & Humida, 1989). In recent times, it was stated that the hibiscus extract can negatively affect the male rat reproductive system (Ali et al., 2012; Sirag, Ahmed, Algaili, Mohamed, & Tajeldeen, 2013). As hibiscus preparation often promote and utilized as agents for improving health, their possible toxic effects and drug interactions should experience careful assessment (Da-Costa-Rocha et al., 2014; Nunes et al., 2017). This chapter aims at providing the most salient recent information on the toxicity and hibiscus
drug interaction.

12.2 Hibiscus
drug interactions

12.2 Hibiscus
drug interactions Hibiscus infusions have long been used to treat many diseases in humans; however, the effects of hibiscus and hibiscus-containing products still need comprehensive pharmacological studies (Da-Costa-Rocha et al., 2014). In recent years, the growth in sales of hibiscus and hibiscus containing products has directed many scientists to turn their research to the pharmacological mechanisms underlying herb
drug interactions. Natural products
drug interactions arise when the pharmacokinetic profile of either product is changed considerably, therefore their coadministration leads to increased adverse effects, toxicity, or therapeutic failure (Izzo, Hoon-Kim, Radhakrishnan, & Williamson, 2016; Johnson et al., 2013). Hibiscus products contain valuable ingredients that could promote human health. The question is to what extent can they be good or unsafe? In addition, they may interact with artificial drugs and increase their toxicity or reduce their concentration to subtherapeutic margins (Jacquin-Porretaz et al., 2017). For instance, the coadministration of hydrochlorothiazide with methanolic extract hibiscus (40 mg/kg) in adult albino mice resulted in a substantial increase of the urinary secretion along with a substantial reduction in the pH value of urine and the levels of sodium, bicarbonate, and chloride ions (Ndu, Nworu, Ehiemere, Ndukwe, & Ochiogu, 2011). The authors suggested that patients who use hydrochlorothiazide diuretics in the management of hypertension should avoid hibiscus containing beverages. The same authors observed the interaction of hibiscus with hydrochlorothiazide in adult rabbits. Overall, the results of these experiments showed that the extracts might lead to retention, and slower removal and interaction were directly associated with increased doses of extract (Hopkins et al., 2013; Ndu et al., 2011). Mahmoud (1994) evaluated the pharmacokinetics of chloroquine (600 mg) and hibiscus beverage in healthy males (N 5 6). The results presented a substantial decrease in the area under the peak plasma concentration against the time curve and the peak plasma level of chloroquine. Similarly, interactions with acetaminophen were also observed (Kolawole & Maduenyi, 2004). Considering the very small sample size (N 5 6) and the inadequate information on the method of preparation and dosage of the hibiscus extracts, the outcome of these studies should be interpreted carefully. Recently, Johnson et al. (2013) studied the in vitro interaction of ethanolic hibiscus extract and cytochrome p450 isoforms (enzymes accountable for the oxidation of the various and complex array of drugs and foreign chemicals to which the body is exposed). At a level of 306
1660 μg/mL, the preparation presented 50% inhibition of the isoforms of P450. These findings indicated that attention should be paid to the simultaneous consumption of hibiscus along with drugs. Conversely, Prommetta, Phivthong-Ngam, Chaichantipyuth, Niwattisaiwong, and Lawanprasert (2006) in their in vivo experiment on the inhibitory effect of hibiscus extract on cytochrome P450 (CYP) indicated that this might not lead to substantial herb
drug interaction.

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Jacquin-Porretaz and coauthors observed an interaction between erlotinib (Erlotinib is a tyrosine kinase receptor inhibitor) and hibiscus tea in a woman with Non-small cell lung carcinoma treated with erlotinib for 5 years. The patient developed an acute cutaneous toxic effect unexpectedly developed on account of daily self-administration of hibiscus tea (Jacquin-Porretaz et al., 2017). Again, the cessation of hibiscus tea together with relevant treatments, including ciclopiroxolamine and clobetasol on the scalp and topical bethametasone on the skin lesions resulted in fast regression of her cutaneous lesions in 5 days. The patient was able to continue administering erlotinib (Jacquin-Porretaz et al., 2017). This case suggested that the nurses, pharmacists, and patients should be conscious of the possible interactions between newly developed oral antineoplastic drugs and medicinal plants such as hibiscus to reduce serious adverse effects. Until recently, the specific mechanism of herb extracts that affect the medicine’s absorption, distribution, metabolism, and excretion properties is not well understood (Liu et al., 2015). However, it is suggested that both pharmacokinetic and pharmacodynamic components of drugs can be altered by coadministered herbs (Fig. 12.1). Overall, the currently available data from preclinical, clinical, and case report studies provide substantial evidence of possible hibiscus extract
drugs interaction.

FIGURE 12.1 Pharmacokinetic and pharmacodynamic mechanisms for drug
herb interactions. Copyright (2007), with permission from Elsevier.

12.3 Hibiscus sabdariffa toxicology

12.3 Hibiscus sabdariffa toxicology In recent years, the utilization of complementary and alternative medicines produced from hibiscus in the medication of several diseases has grown. Possibly toxic chemicals incorporated into those preparations to increase their expected impacts or the unconscious utilize of hibiscus tea have resulted in some toxic effects. The study conducted on the animal has primarily demonstrated that excessive doses of hibiscus infusions for relatively long term might have detrimental effects on the testicles of experimental animals (Ali, Wabel, & Blunden, 2005). The LD50 of hibiscus infusion in rats was considered 5000 mg/kg. The aqueous extract of hibiscus calyx (3%) was administered orally for up to 4 weeks at a dose of 200 mg/kg body weight/mouse (Mahmoud, 2012). The albino rats were beheaded and the testes and epididymides were removed and prepared for transmission electron microscopy to evaluate ultrastructural and semen defects. The author found that both cold and boiled water extracts from hibiscus calyx affected sperm morphology and testicular ultrastructure and adversely affected the male reproductive system in mice. Ali et al. (2012) reported on the adverse effect of hibiscus (10%, 15%, and 20%) administered for 70 days and its anthocyanins (50, 100, and 200 mg/kg) for 5 days on the weight and histology of the testis, and on some biochemical parameters in testicular samples. Furthermore, the plasma levels of testosterone, luteinizing hormone (LH), and estradiol were measured. The potential existence of an estrogenic effect of hibiscus and anthocyanins on the uteri of immature female rats was also investigated. The authors noted both the hibiscus extract and anthocyanins significantly altered either testicular weight or uterus weight but both hibiscus and anthocyanins showed normal testicular histology. In another study, testicular damage in male Wistar rats was evaluated (Olusanya, Rasaq, & Ojemekele, 2018). In this study, hibiscus treatment significantly decreased the plasma levels of the reproductive hormones testosterone, estradiol, prolactin, and LH, and follicle-stimulating hormone (FSH) in comparison to the control group. Histopathological investigation of the testes also revealed marked degeneration of the seminiferous tubules, with necrosis and alteration in testicular structure. Similar testicular toxicological effects in the old male Wistar albino rats were also observed when hibiscus extract was orally administered at doses of 1.15, 2.30, and 4.60 g/kg for 12 weeks (Orisakwe et al., 2004). The workers hypothesized that these effects were associated with the intervention by the extract with spermatogenesis that might be triggered by an estrogenic activity of the extract. Indeed, some work has previously presented this possibility (Ali et al., 1989). On the other hand, Amin and Hamza (2006) observed that the consumption of a low dose of hibiscus (1 g/kg/day) extract administered to experimental animals for 26 days improved the actions of testicular antioxidant enzymes and restored sperm motility of cisplatin-treated male albino rats. In another study, the effect of ethanol of hibiscus calyx extracts on reproductive hormones in males and females of rats was evaluated (Sirag et al., 2013). The author observed that the

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administration of hibiscus extract (250 mg/kg) induced slight implications on rat reproductive hormones, specifically testosterone and estradiol, whereas no significant change was found on the levels of prolactin and FSH. Furthermore, no histological alterations were observed on both testes and ovaries after 28 days of administration. The author concluded that the ethanolic extract of hibiscus at a dose of 250 mg/kg induced minor effects on rat reproductive hormones. Onyenekwe et al. (1999) investigated the antihypertensive and adverse effects of the aqueous extract of hibiscus on mice. In this study, no mortalities were reported in mice after 14 days of administering doses of 1000
5000 mg/kg b. w. d. Thus the calculated LD50 of hibiscus extract was greater than 5000 mg/kg b. w. The same researchers evaluated the impact of the hibiscus extract on blood pressure in spontaneously hypertensive and normotensive rats. Administration of the highest dose of the extract (1000 mg/kg) during the 7 and 21 days caused deaths in spontaneous hypertensive but not in normotensive rats. Based on the well-known greater risk of sudden cardiac death in patients sick administering non-potassium sparing diuretics, the researcher attributed deaths to a diuretic result of the extract (Onyenekwe et al., 1999). Ebenezer and coauthor evaluated the toxicological effects for 8 weeks’ of various doses (250, 500, and 1000 mg/kg b. w.) of hibiscus leaves in adult male Wistar rats. The results showed strong testicular toxicity effects (decrease sperm amount, motility, and viability with an indication of obvious degenerative histological changes) in all 1000 mg/kg group compared with the control (Ebenezer, David, Koyinsola, Olayemi, & Olutoyi, 2019). Following this study, another clinical experiment of aqueous extract hibiscus on mice at doses 50 and 100 mg/kg. b. w. for 21 days showed a significant reduction in the testis weight, sperm counts, sperm motility, and viability in the experimental animals (Arabi, Kahil, & Ibrahim, 2014). Additionally, the study revealed that there was a significant difference in LH, FSH, and testosterone hormone in the treatment groups. The authors suggested that hibiscus has an adverse effect on sperm parameters of mice. de Arruda, Cardoso, Vieira, and Arena (2016) have investigated the developing reproductive system of rats whose mothers were treated with hibiscus extracts (250 and 500 mg/kg) after 12 days of pregnancy up to day 21 of lactation, with the purpose of finding information concerning the reproductive safety of hibiscus extracts. At puberty, a significant decrease in the sperm count in the caput/corpus of epididymis after administration of both doses and a decline in the sperm count in the cauda epididymis for 250 mg/kg dose were observed. The authors suggested that maternal exposure to hibiscus extract can negatively impact the male reproductive system in rats. Additionally, there were adverse effects on the volume and semen characteristics of the Sudanese Desert when hibiscus seeds (30%) were administered (Ali et al., 2014). Sirag et al. (2013) administered a dose of 250 mg/ kg of ethanolic extracts of hibiscus on rats (male and female) reproductive hormones. The hibiscus extract revealed mild effects on the rat reproductive system, mainly testosterone and estradiol, whereas no significant alteration was detected on the levels of FSH and prolactin. Furthermore, no biological alterations were observed on both testes and ovaries of rats during 28 days of administration.

12.4 Effect of hibiscus on kidney and liver functions

12.4 Effect of hibiscus on kidney and liver functions Fakeye et al. (2009) evaluated the toxicity and immunomodulatory effects after 90 days of administration of 50% alcoholic extracts of hibiscus at doses of 300 and 2000 mg/kg b. w. in rats, respectively. By day 8, the author observed severe toxicity with total mortality in the highest dose in both water and alcohol extract
treated groups, whereas 50% ethanol extract group at 2000 mg/kg until 28 days and in the water and 50% alcoholic groups at 300 mg, until day 60 and 40, respectively. A significant increase of plasma creatinine was detected when hibiscus extract (300 mg/kg b. w.) was administered for 30 days. High creatinine blood level could be attributed to muscular dystrophy, loss of kidney function, or even death. Although the alcoholic extract showed a great increase in creatinine without death at 300 mg dose level, it is questionable to attribute the reason for deaths to the higher level of creatinine (Fakeye et al., 2009). Ukoha, Mbagwu, Ndukwe, and Obiagboso (2015) evaluated the effect of hibiscus infusions (200
800 mg/kg b. w.) on Wistar rats for 21 days. The results showed that the long-term consumption of hibiscus could affect kidney function by causing a distortion in the integrity of the kidney cytoarchitecture and induce notable increment in serum urea, creatinine, and certain electrolytes which might impair normal kidney function. It has been found that hibiscus extract is rich in sodium ions (Na1) indicating that its oral administration increases plasma concentration (Iyare, Adegoke, & Nwagha, 2010). The mechanism for the sodium increase might be attributed to the activity of flavonoid compounds in the hibiscus (Emelike & Dapper, 2013). Generally, the findings of this study are in agreement with the literature (Kirdpon, Nakorn, & Kirdpon, 1994; Orisakwe et al., 2004). Akindahunsi and Olaleye (2003) noticed the toxicological effects of methanolic extract (250 mg/kg) of hibiscus when administered to rats at 15 doses. This study indicated that the level of serum aspartate aminotransferase and alanine aminotransferase was substantially increased in all treated animals. However, the serum concentrations of alkaline phosphatase and lactate dehydrogenase were not substantially changed. Among the treatments, the group with 15 doses showed a significant increase in their serum level of albumin. From the above results, it is obvious that long-term consumption of alcoholic extract of hibiscus at a 15-dose level may induce liver damage, whereas the effect was slight at small dose levels (1
10). Therefore the author recommended that the consumption of 150
180 mg/kg/day is safe and the hibiscus extract must be consumed with caution since higher doses might affect the liver functions. The water extract of hibiscus (0.6 g/100 mL and 8 g/100 mL) was administered orally to rats for up to 28 days. The results suggested that the long-term administration of this extract could lead to an increase in kidney and liver enzymes which might have been accountable for the significantly decreased food consumption (Emelike & Dapper, 2013). Prommetta et al. (2006) investigated the effect of hibiscus at doses of 0.25 and 1.00 g/kg/day for 30 days on rats and no significant toxic effect was observed

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on blood chemistry and hematology and liver. Similarly, Okasha, Abubakar, and Bako (2008) administered hibiscus extract in a dose of 1.0 for 4 weeks and did not observe any substantial alterations in the liver or renal function of the experimental animals. The diversity of the results of many researchers could be due to the variations in the concentration and the bioactivity of anthocyanins administered to the experimental group.

12.5 Other adverse effects Several studies that demonstrated the effect of hibiscus tea on reproduction and development in animal models have been reported (Iyare & Adegoke, 2008a, 2008b). In a study, the effect of administering hibiscus extract to pregnant and lactating rats on pregnancy outcomes and postnatal was investigated (Iyare, 2010). From this study, it can be concluded that maternal consumption of aqueous extract of hibiscus during pregnancy and lactation could delay the onset of puberty. In another study, the administration of hibiscus extract to the female rats caused a dosedependent delayed onset of puberty (Iyare & Adegoke, 2008a, 2008b). The same authors investigated the mechanism of the delayed puberty onset in the offspring of rats that were treated with aqueous extract of hibiscus extract (0.6 g/100 ml and 1.8 g/100 ml) during pregnancy of Sprague-Dawley rats (Iyare et al., 2010). The authors observed that the administration of hibiscus increased postnatal weight gain, delayed puberty onset, and raised body mass index at the onset of puberty. The study suggested that these effects might be attributed to an increase in the maternal plasma ion (Na 1 ) and corticosterone (is a steroid hormone of the adrenal cortex) levels during pregnancy. These studies required more validation as there are no observations that have been reported in humans. Malik et al. (2013) investigated the aluminum and other elements in hibiscus infusions. A high quantity of available aluminum (272 6 19 mg/kg) could have a negative impact on human health. According to a potentially harmful level of aluminum, the hibiscus infusion should not be consumed in the amount of greater than 1 L/day (total aluminum allowance up to 1.2 6 0.1 mg/L) by vulnerable groups including pregnant women, and should be avoided from the food of children below 6 months of age and children with chronic renal failure.

12.6 Conclusion Hibiscus infusions and products containing hibiscus (mainly the preparation and aqueous extracts) have a long-term conventional utilization both in food and medication and generally are deemed to be safe. The database searches for this chapter reveal a well-documented case report, preclinical and clinical studies of toxicity and adverse reactions resulting from oral consumption of hibiscus

References

preparations. Several studies have demonstrated that hibiscus tea consumption has low toxicity when consumed at low doses without any toxic effect on the reproductive systems and liver or kidney functions. However, additional studies are required to establish a prospective approach that can balance the pharmacological and toxic properties of hibiscus preparation. The bioavailability and dosage of hibiscus extract are still an additional subject that has to be studied. Hereafter, there is a requirement for the standardized fingermark of hibiscus worldwide for quality control. Comprehensive knowledge of the mechanism of hibiscus
drug interaction is crucial for clinical risk evaluation, in turn, crucial to health professionals in their efforts to lessen risk and guarantee that consumption of hibiscus and hibiscus-containing products are as safe as possible.

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CHAPTER

Conventional and rapid methods for measurement of total bioactive components and antioxidant activity in Hibiscus sabdariffa

13

Huang Xiaowei1,2,*, Li Zhihua1, Haroon Elrasheid Tahir1, Zou Xiaobo1, Shi Jiyong1, Xu Yiwei1 and Zhai Xiaodong1 1

School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China School of Agricultural Equipment Engineering, Jiangsu University, Zhenjiang, P.R. China

2

Chapter Outline 13.1 Introduction ...............................................................................................199 13.2 Spectrophotometric techniques ...................................................................201 13.2.1 Conventional measurement of total phenolic content ..................201 13.2.2 Measurement of total flavonoid content and total anthocyanin content ..................................................................................204 13.2.3 Measurement of antioxidant activities .......................................204 13.3 Near-infrared spectroscopy .........................................................................205 13.4 Chemometrics analysis ...............................................................................206 13.5 Determination of phenols, flavonoid, anthocyanins, and antioxidant activities in Roselle ..................................................................................................207 13.6 Conclusion .................................................................................................209 References ..........................................................................................................210

13.1 Introduction Hibiscus sabdariffa (Hs), an annual shrub, is commonly used to make jellies, jams, and beverages. The brilliant red color and unique flavor make it a valuable * Contributed equally to the work. Roselle (Hibiscus sabdariffa). DOI: https://doi.org/10.1016/B978-0-12-822100-6.00011-2 © 2021 Elsevier Inc. All rights reserved.

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CHAPTER 13 Conventional and rapid methods for measurement

food product (Jabeur et al., 2017; Tsai, McIntosh, Pearce, Camden, & Jordan, 2002). Hs is an important cash crop grown on the East Coast of Malaysia, south of China, India, and West Africa (Dhar, Kar, Ojha, Pandey, & Mitra, 2015). The anthocyanin pigments that create the color are responsible for the wide range of coloring in many foods (Para´ıso et al., 2019). Being high in anthocyanin, Hs petal is both a good colorant and potentially a good source of antioxidants. It is also rich in bioactive compounds such as anthocyanins and other flavonoids, organic acids, and polysaccharides which are responsible for its antioxidant, antibacterial, antiinflammatory, hepatoprotective, and anticholesterol activities (Ismail, Ikram, Hainida, Nazri, & Saadiah, 2008; Shahidi & Ambigaipalan, 2015). In developing countries a large section of the population relies on medicinal plants for primary health care requirements (Da-Costa-Rocha, Bonnlaender, Sievers, Pischel, & Heinrich, 2014). Traditional medicines are becoming popular among most of the world population mainly because they are cheap, abundant with less adverse effects on health. In recent years, focus on plant research has increased globally to find out the immense potentials of medicinal plants used in various traditional systems. Various medicinal plants have been studied which could be used as potent phytochemical agents in the therapeutic treatment of various diseases; one among them is Hs, known for its delicacy and medicinal properties which has several health benefits (Riaz & Chopra, 2018). As the main bioactive and antioxidant components of Hs, anthocyanin is measured by some common chemical methods, such as high-performance liquid chromatography and pH differential method (Chensom et al., 2020; Ongkowijoyo, Luna-Vital, & de Mejia, 2018). These methods are precise but destructive, time-consuming, and costly. It is necessary to explore a rapid and conventional method for the measurement of bioactive components in Hs (Alam, Bristi, & Rafiquzzaman, 2013; Granato et al., 2018; Man, 2002). In recent years, near-infrared spectroscopy (NIRS) has gained wide acceptance in different fields by virtue of its advantages over other analytical techniques, the most salient of which is its ability to record spectra for solid and liquid samples without any pretreatment (Slocombe, Ross, Thomas, McNeill, & Stanley, 2013). This characteristic makes it especially attractive for straightforward, speedy characterization of natural and synthetic products. The cost savings of NIR measurements related to improved control and product quality are often achieved and can provide results significantly faster compared to traditional laboratory analysis (Tahir, Zou, Shi, Mariod, & Wiliam, 2016). NIRS is a noninvasive, simple, quick, and inelastic scattering technique that can detect the structural characterization of substances on molecular levels (Arslan et al., 2018). Applications of NIR for the determination of biological active compounds and antioxidants in food are of great interest and have gained great attention in recent years, due to their rapidity and low cost (Tahir et al., 2017, 2019). NIR has been used for the determination of bioactive components, antioxidant activity, and macronutrients in fruits (Kljusuri´c, Jurina, Valinger, Benkovi, & Tuˇsek, 2020), black goji berries (Arslan et al., 2018), isoflavone and saponin content of ground

13.2 Spectrophotometric techniques

soybeans (Berhow et al., 2020), phenolics and methylxanthines in spent coffee grounds (Magalha˜es, Machado, Segundo, Lopes, & Pa´scoa, 2016), active constituents and antioxidant capability of green tea (Guo et al., 2020), bioactive compounds in intact cocoa bean husk (Herna´ndez-Herna´ndez, Ferna´ndez-Cabana´s, Rodr´ıguez-Gutie´rrez, Bermu´dez-Oria, & Morales-Sillero, 2021), bioactive of freeze-dried acai (Carameˆs, Baqueta, Conceic¸a˜o, & Pallone, 2021), phytochemical composition and variability in Quercus ilex acorn morphotypes (Lo´pez-Hidalgo, Trigueros, Mene´ndez, & Jorrin-Novo, 2021), anthocyanins in wine (RomeraFerna´ndez et al., 2012), and polyphenol and antioxidants activity in Chinese Dates (Zizyphus jujuba Mill.) (Arslan, Xiaobo, et al., 2019; Arslan, Zou, et al., 2019). This chapter will discuss the current state of the study on spectroscopies applications for measurement of bioactive (total phenol, total flavonoids, and anthocyanins) and antioxidant activities in Roselle. Our main focus will be on the NIR technique due to its wide adaptability and application in the food industry (Rodriguez-Saona, Giusti, & Shotts, 2016). We will also present the available research on the UV/Vis spectroscopic application as one of the spectroscopic widely used in analysis of food because of its ease of usage and nondestructive (Aleixandre-Tudo, Nieuwoudt, Olivieri, Aleixandre, & du Toit, 2018; Lopes, Moresco, Peruch, Rocha, & Maraschin, 2017; Sukwattanasinit, Burana-Osot, & Sotanaphun, 2016). The objective of this chapter was to introduce conventional and rapid methods for the measurement of total bioactive components and antioxidant activity in Roselle.

13.2 Spectrophotometric techniques The spectrophotometric technique is the conventional method used for the measurement of total polyphenols in foods. It is depending on the reaction of a radical, radical cation, or complex with biologically active materials molecule able to transfer a hydrogen atom (Pisoschi & Negulescu, 2011).

13.2.1 Conventional measurement of total phenolic content The colorimetric method based on the reaction of Folin Ciocalteu reagent (FCR) is commonly applied for the measurement of total phenolic content (TPC) in Roselle (Andzi Barhe´ & Feuya Tchouya, 2016; Mohd-Esa, Hern, Ismail, & Yee, 2010; Sindi, Marshall, & Morgan, 2014). The assay is composed of calibration with a pure standard of phenolic compound (mostly gallic acid), extraction of phenol from the sample, and recording the absorbance after the reaction. The sample preparation methods are presented in Table 13.1. The main drawback of this method is low specificity because this reagent (FCR) does not only quantity total phenols, but also with any reducing agent and the color reaction can take place with any oxidizable phenolic hydroxy group such as aromatic amines,

201

Table 13.1 Representative spectrophotometric methods for the determination of bioactive compounds and antioxidant activities in Roselle. Method of preparations

Parameters measured

References

The dried calyx powder (1 g) was dissolved in 50 mL water and sonicated (continuous, 20 kHz) for 20 min at 25 C. The freeze-dried calyces (1 g) were extracted with alcohol and then evaporated at 40 C under vacuum. The dried calyces (25 g) were dissolved in ultrapure water and stirred in a vortex until dissolved.

The aqueous extract was used for the determination of TPC, TFC, total antioxidant content, FRAP, and DPPH% The extract was used to determine the total phenolics and anthocyanin The aqueous extract of Roselle was used for the determination of TEAC, FRAP, ORAC, and measurement of TBARS The extracts were for measurement of total polyphenol content, antioxidant capacity using DPPH, FRAP, and TEAC assays.

Tahir, Xiaobo, et al. (2016)

The extract was used for the analysis of antioxidant activity using ABTS, TMA, and TPC.

Cassol, Rodrigues, and Zapata Noreña (2019)

The aqueous extracts were used to analyze TAC, TPC, and ORAC

Ramirez-Rodrigues, Plaza, Azeredo, Balaban, and Marshall (2011) Ifie, Ifie, Ibitoye, Marshall, and Williamson (2018)

The dried calyces powder (0.1 g) were extracted using various solvent (water, methanol, ethyl acetate or hexane; 10 mL) with and without the addition of 1% (v/v) formic acid solution. Then, the extracts of each solvent were heated at 25 C, 50 C, or at the solvent boiling point for either 3, 5, or 10 min. Before extraction, the calyces in the ratio of 1:5 (calyx: acidified water) were steam water at 100 C for 4 min and then acidified water (2% citric acid, w/v) was added, with pH B2.1, to improve the stability of anthocyanins for the stability of the anthocyanins. The dried calyces were dissolved in the ratio 1.4 (calyxto-water ratio) and then the extracts were heated at 25 C for 240 min and 90 C for 16 min. The dried calyx powder (2 g) was dissolved in 1000 mL distilled water and heated in the water bath at 50 C for 30 min with intermittent stirring. Then, the mixture was centrifuged (2500 g; 10 min), filtered through a Whatman no. 1 filter paper, and used for the analysis. The extraction process was performed three times.

The aqueous extract was used to determine TPC

Christian and Jackson (2009) Fernández-Arroyo et al. (2011) Sindi et al. (2014)

Different parts of Roselle (calyces, stems, and leaves) were dissolved in a ratio of 1:1000 (calyx-to-water ratio) with distilled water or 80% (v/v) methanol for 2 h at room temperature, using an orbital shaker The mixture was filtered through a filter paper. The dried calyces (2.5 g) were mixed with 10 mL of distilled water using homogenizer and then heat at 95 C in a water bath for 1 h and then centrifuged at 11,000 rpm and allowed to cool at room temperature for 7 min. The extraction process was repeated until the volume of the supernatant reached 25 mL. The combined extracts were filtrate and used for further analysis. Dried leaves (0.5 g) were blended with 25 mL deionized water. The mixture was kept at room temperature in the dark place for 1 h, with occasional agitation. Then, the mixture was filtered using filter paper (Whatman No. 1).

The extract was used to estimate antioxidant activities using the β-carotene bleaching

Mohd-Esa et al. (2010)

The extract was used for evaluation of total anthocyanin content

Sukkhaeng, Promdang, and Doung-ngern (2018)

The water extract was used for the determination of TPC, TEAC, DPPH, FRAP, and CCA

Wong, Leong, and William Koh (2006)

ABTS, 2,20 -Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid); CCA, cupric ion chelating activities; DPPH, 1,1-diphenyl-2-picrylhydrazyl; FRAP, ferric reducing antioxidant power; ORAC, oxygen radical absorbance capacity; TAC, total anthocyanin content; TBARS, thiobarbituric acid reacting substances; TEAC, Trolox equivalent antioxidant capacity; TFC, total flavonoid content; TMA, total monomeric anthocyanins; TPC, total phenolic content.

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CHAPTER 13 Conventional and rapid methods for measurement

ascorbic acid, Cu(I), and Fe(II) (Alvarez-Suarez, Tulipani, Romandini, Vidal, & Battino, 2009). For the above reasons, many scientists believe that it is not appropriate for measuring total phenol until interfering agents such as sugars are considered or removed (Prior, Wu, & Schaich, 2005; Singleton, Orthofer, & Lamuela-Ravento´s, 1999).

13.2.2 Measurement of total flavonoid content and total anthocyanin content The quantitative measurement of flavonoids in the Roselle is possible by spectrophotometric technique. The aluminum chloride technique is applied to measure the flavone and flavonol content; it is mechanism is the formation of a composite between the aluminum ion Al(III) and the carbonyl and hydroxyl groups of the flavonoid (Markham, 1982). Colorimetric methods for the determination of total anthocyanins using pH differential procedures are based on their characteristic behavior under acidic solutions. The basis of this assay is the reduction of pH to values ranging between 0.5 and 0.8 which causes all anthocyanins to transform to red-colored flavylium cation (Lapornik, Proˇsek, & Wondra, 2005). Spectrophotometric assays give very valuable quantitative and qualitative information. Generally, spectrophotometric is the main technique used for the measurement of different groups of polyphenols due to its ease of use and inexpensive (Ignat, Volf, & Popa, 2011). Detailed information of procedures used in the characterization and quantification of anthocyanins by UV
Vis can be found in the literature (Giusti & Wrolstad, 2003).

13.2.3 Measurement of antioxidant activities Spectrometric methods depend on the reaction of the radical, radical cation, or complex with an antioxidant molecule capable to donate a hydrogen atom (Pisoschi & Negulescu, 2011; Shahidi & Ambigaipalan, 2015). The need to measure the antioxidant capacity of Roselle is well reported; these provided important information to food processors and consumers. Roselle plants are rich in antioxidant compounds such as phenolic acid, flavonoids, ascorbic acid, and anthocyanins (Da-Costa-Rocha et al., 2014; Riaz & Chopra, 2018). There are many procedures for evaluating the antioxidant capacities in Roselle samples (Table 13.1); these can be divided into two groups. The first group measures the capability of antioxidants in preventing oxidation in a model system by monitoring the associated changes using physical, chemical, or instrumental means (Shahidi & Ambigaipalan, 2015). Radical scavenging methods include assays connected with hydrogen atom transfer (HAT) or single electron transfer (SET) mechanisms, such as β-carotene bleaching, oxygen radical absorbance capacity, cupric reducing antioxidant capacity (CUPRAC), and 2,20 -azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), while Trolox equivalent antioxidant capacity, ferric

13.3 Near-infrared spectroscopy

reducing antioxidant power (FRAP), and 1,1-diphenyl-2-picrylhydrazyl (DPPH) method represent SET-based assays (Shahidi & Ho, 2007). HAT-based assays evaluate the ability of biologically active compounds to suppress free radicals via hydrogen donation, while SET-based assays evaluate the capacity of a potential of biologically active compounds to transfer one electron to reduce any components such as metal, carbonyl, and radical. Detailed information of these assays is beyond the aim of this chapter and can be found elsewhere (MacDonald-Wicks, Wood, & Garg, 2006; Pisoschi & Negulescu, 2011; Shahidi & Ambigaipalan, 2015).

13.3 Near-infrared spectroscopy In recent years, NIR has been imposed as a rapid analysis used for quantitative determination of photochemical components of a wide of agricultural products (Tahir et al., 2019). NIR is a region that presents the molecular bond vibrations, in the spectral zones between range 14,000 and 4000 cm21, offering valuable information about the physical and chemical features of samples. The NIR spectrum results from complex overtones and combination of tones (Lee, 2007; Ozaki, McClure, & Christy, 2006). The appropriate mode of NIR showed selected according to the optical properties of the food samples (Fig. 13.1). NIR technique is based on the absorbance of radiation at molecular vibrational frequencies occurring for the O
H, N
H, S
H, and C
H groups (Chadha & Haneef, 2015). According to the equipment, the spectrum of the analyzed sample will generate a large data set. Thus multivariate analyses are employed to extract useful information and reduce noises and background information.

FIGURE 13.1 Infrared spectroscopy assessment modes—(A) transmittance, (B) diffuse transmittance, (C) diffuse reflectance, and (D) transflectance (Tahir et al., 2019).

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13.4 Chemometrics analysis The term chemometrics can be described as the science of extracting information from chemical structures via mathematical modeling of data (Wold, 1972). In 1997 Massart defined chemometrics as a “chemical discipline that utilizes mathematics and formal logic in order to (1) design optimum experimental methods; (2) to produce maximum chemical information by evaluating chemical data; and (3) to get information regarding chemical systems” (Massart et al., 1998). In this chapter, a brief introduction to supervised methods used in the determination of bioactive in the Roselle has been presented. Other organizations that transcend the scope of this work were discussed elsewhere. Data analysis of spectra consists of spectral data pretreatment, developing a calibration model, and model transfer (Corte´s, Blasco, Aleixos, Cubero, & Talens, 2019). Spectra pretreatment techniques can be used for removing the background and reducing the noises as presented in Fig. 13.2

(A)

(B)

Wavenumbers (cm-1)

Wavenumbers (cm-1)

(C)

Wavenumbers (cm-1)

FIGURE 13.2 NIR spectra of Hibiscus samples obtained from (A) raw spectra, (B) SNV, and (C) MSC preprocessing spectra (Tahir, Xiaobo, et al., 2016). MSC, Multiplicative scatter correction; NIR, near-infrared; SNV, standard normal variate.

13.5 Determination of phenols, flavonoid, anthocyanins

(Tahir, Xiaobo, Tinting, Jiyong, & Mariod, 2016). Several preprocessing techniques such as standard normal variate (SNV) transformation, multiplicative scatter correction (MSC), mean centering (MC), derivative algorithm, baseline correction, and smoothing algorithms have been employed. Implementation of a calibration model on such preprocessed data might result in a good model. Due to the huge overlap and complex nature and the complex nature of continuous data, occasionally, it is difficult to figure out the positions of specific bands that characterize the different components in agricultural products. To avoid using unimportant data, the selection of feature wavelengths in the NIR region is a very effective technique to establish a robust and accurate model. Wavelength selection methods such as Ant colony optimization interval partial least squares (ACO-iPLS) and genetic algorithm interval partial least squares (GA-iPLS) were used to develop calibration models for total anthocyanins content (Huang et al., 2014). Partial least squares regression (PLSR) is one of the most common modeling approaches for the quantitative determination of bioactive and antioxidant activities in food (Tahir, Xiaobo, et al., 2016; Tahir et al., 2019). Model accuracy is evaluated using correlation coefficient (R), lower error values in RMSECV (root mean square error of the cross validation), RMSEP (root mean square error of prediction) and ratio performance deviation (RPD) ratio. RPD was calculated as the ratio of the standard deviation of the response variable to the standard error of prediction (RMSEP).

13.5 Determination of phenols, flavonoid, anthocyanins, and antioxidant activities in Roselle Tahir, Xiaobo, et al. (2016) investigated the application of NIR for quantitative analysis of TPC and total flavonoid compound content (TFC) in Roselle samples. The spectra were obtained using about 2 g of Roselle fine powder. The samples were analyzed in triplicate and the mean of the triplicate spectra which was acquired from the identical samples was used for chemometric analysis. To improve the accuracy and robustness of the model, the spectral was preprocessed by SNV, MSC, MC, and baseline correction, and the prediction model was constructed using PLSR. As presented in Table 13.2, the pretreated spectra using SNV can provide more information corresponding to TFC, while the MSC-treated spectra were more suitable to measure the TPC in Roselle powder. This could indicate that the TPC and TFC were related to different useful information in the spectra (Huang, Wu, et al., 2011). Similar performance of SNV and MSC were obtained when they applied to extract information associated with bioactive compound and antioxidant activity (Escuredo, Carmen Seijo, Salvador, & Inmaculada Gonza´lez-Mart´ın, 2013; Moncada, Gonza´lez Mart´ın, Escuredo, Fischer, & M´ıguez, 2013). The results of PLSR models showed the NIR technique effective and enabled the quantitative determination of TPC and TFC with high correlation coefficients 0.86 and 0.83, respectively.

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Table 13.2 Prediction results of antioxidants and antioxidant activity of Hs by partial least squares with different preprocessing methods in calibration and prediction analysis. Parameters

TPC

TFC

TAC

FRAP

DPPH%

Preprocessing

SNV MSC MC Baseline SNV MSC MC Baseline SNV MSC MC Baseline SNV MSC MC Baseline SNV MSC MC Baseline

Calibration set (N 5 48)

Validation set (N 5 24)

Rc

RMSECV

Rp

RMSEP

RPD

0.87 0.84 0.81 0.85 0.85 0.78 0.77 0.86 0.91 0.84 0.80 0.79 0.91 0.70 0.87 0.69 0.87 0.82 0.82 0.85

0.39 0.29 0.31 0.30 0.08 0.12 0.10 0.09 0.16 0.20 0.19 0.21 0.33 0.46 0.39 0.59 0.29 0.34 0.33 0.30

0.82 0.86 0.76 0.79 0.83 0.73 0.75 0.67 0.91 0.83 0.80 0.80 0.89 0.61 0.82 0.66 0.86 0.78 0.75 0.80

0.47 0.24 0.31 0.32 0.14 0.09 0.13 0.16 0.08 0.16 0.22 0.20 0.34 0.72 0.49 0.55 0.29 0.36 0.40 0.41

2.20 3.37 1.79 1.92 8.25 2.31 1.40 1.40 4.19 2.42 1.38 1.95 3.30 1.03 1.73 1.56 5.10 1.76 1.40 1.51

DPPH, 1,1-Diphenyl-2-picrylhydrazyl; FRAP, ferric reducing antioxidant power; MC, mean centering; MSC, multiplicative scatter correction; Rc, correlation coefficients of calibration; RMSECV, standard error of cross-validation; Rp, correlation coefficients of prediction; RPD, ratio (SD/SEPC) performance deviation; SNV, standard normal variate; TAC, total anthocyanin content; TFC, total flavonoid content; TPC, total phenolic content.

The RPD values of TPC and TFC were 3.37 and 8.25, respectively (Table 13.2). Excellent calibration and validation models were established for TPC and TFC with high ability of prediction (RPD . 3), indicating the potential use of the NIR technique as a rapid and nondestructive method for the determination of bioactive compounds in Roselle calyces. In another study, Sukwattanasinit et al. (2016) evaluated the total anthocyanin content (TAC) and TPC in Roselle using UV/Vis spectroscopy (190
1100 nm) in combined with PLSR and multiple linear regression (MLR) models established from the absorbance at 330, 380, and 530 nm. The accuracy of PLS and MLR models was revealed by the low root-mean-square error of prediction 5 0.11
0.13 and a high correlation between the reference method and

13.6 Conclusion

prediction values of the analyzed samples (R 5 0.99). These results indicated that UV/Vis spectroscopy combined with the chemometrics was reliable, simple, and fast and could be applied in routine work. Xiaowei et al. (2014) evaluated the feasibility of NIR spectra (10,000
4000 cm21) for rapid measurement of TAC in the Roselle mixed with the other flowering plants using NIR in conjunction with the ACO-iPLS and GAiPLS. The spectra zone (4590
4783, 5770
5963 cm21) corresponding to TAC were selected by ACO-iPLS. The best ACO-iPLS model for TAC (R 5 0.9856, RMSECV 5 0.1198 mg/g) showed the best prediction model as compared with the performance of full-spectrum PLS, iPLS, and GA-iPLS models. The authors concluded that the NIR technique was accurate for screening the TAC in flowering teas, providing a good indirect representation of TAC. Tahir, Xiaobo, et al. (2016) evaluated DPPH radical scavenging activity (DPPH), FRAP, and total antioxidant content in Roselle using NIR together with the PLSR model. As presented in Table 13.2, the antioxidant activities of test samples were varied, indicating the variations of Roselle calyx’s sources and types (dark red, red) (Tahir, Arslan, et al., 2020; Tahir, Xiaobo, et al., 2016; Tahir et al., 2017). The antioxidant activities measured showed high correlations (0.86
0.91, RPD $ 3.3) and low standard error of cross-validation (0.08
0.33) and root mean standard error of prediction (0.08
0.29). These results indicate that NIR coupled with PLSR can be used for screening of antioxidant activities of the Roselle.

13.6 Conclusion The functional properties of polyphenols and their effect on health increased research efforts to develop a rapid technique for extraction, separation, and quantification of these compounds from natural sources. These techniques must be comprehensive, fast, and rich in spectral information. This chapter provides information on the method of determination of phenolic compounds and antioxidant activity in Roselle, the conventional procedure that is widely used for total phenols, total flavonoid, total anthocyanin, and antioxidant activities, as well as the available rapid procedures that enable further progress in the determination of these compounds. This chapter shows that spectral signals from NIR and UV
Vis combined with the spectral preprocessing method can be used for the direct determination of biological compounds in the Roselle. Furthermore, NIR and UV
Vis techniques are noninvasive, simple, and quick technique, require low operator requirements, and is very suitable for online detection. Future researches have to direct to the unification of preprocessing and optimization of quantitative modeling approaches. Finally, this chapter is important for scientists and researchers, since there is a limited number of researches on the use of a rapid method for the determination of antioxidant compounds in Roselle.

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1060. Moncada, G. W., Gonza´lez Mart´ın, M. I., Escuredo, O., Fischer, S., & M´ıguez, M. (2013). Multivariate calibration by near infrared spectroscopy for the determination of the vitamin E and the antioxidant properties of quinoa. Talanta, 116, 65
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Riaz, G., & Chopra, R. (2018). A review on phytochemistry and therapeutic uses of Hibiscus sabdariffa L. Biomedicine & Pharmacotherapy, 102, 575
586. Rodriguez-Saona, L. E., Giusti, M. M., & Shotts, M. (2016). 4—Advances in infrared spectroscopy for food authenticity testing. In G. Downey (Ed.), Advances in food authenticity testing (pp. 71
116). Woodhead Publishing. Romera-Ferna´ndez, M., Berrueta, L. A., Garmo´n-Lobato, S., Gallo, B., Vicente, F., & Moreda, J. M. (2012). Feasibility study of FT-MIR spectroscopy and PLS-R for the fast determination of anthocyanins in wine. Talanta, 88, 303
310. Shahidi, F., & Ambigaipalan, P. (2015). Phenolics and polyphenolics in foods, beverages and spices: Antioxidant activity and health effects
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897. Shahidi, F., & Ho, C.-T. (2007). Antioxidant measurement and applications: An overview. ACS Publications. Sindi, H. A., Marshall, L. J., & Morgan, M. R. A. (2014). Comparative chemical and biochemical analysis of extracts of Hibiscus sabdariffa. Food Chemistry, 164(0), 23
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178). Academic Press. Slocombe, S. P., Ross, M., Thomas, N., McNeill, S., & Stanley, M. S. (2013). A rapid and general method for measurement of protein in micro-algal biomass. Bioresource Technology, 129, 51
57. Sukkhaeng, S., Promdang, S., & Doung-ngern, U. (2018). Fruit characters and physicochemical properties of Roselle (Hibiscus sabdariffa L.) in Thailand—A screening of 13 new genotypes. Journal of Applied Research on Medicinal and Aromatic Plants, 11, 47
53. Sukwattanasinit, T., Burana-Osot, J., & Sotanaphun, U. (2016). Simple and rapid spectrophotometric method for quality determination of Roselle (Hibiscus sabdariffa). Thai Journal of Pharmaceutical Sciences (TJPS), 40(4). Tahir, H. E., Arslan, M., Mahunu, G. K., Mariod, A. A., Wen, Z., Xiaobo, Z., . . . El-Seedi, H. (2020). Authentication of the geographical origin of Roselle (Hibiscus sabdariffa L) using various spectroscopies: NIR, low-field NMR and fluorescence. Food Control, 114107231. Tahir, H. E., Xiaobo, Z., Jianbo, X., Mahunu, G. K., Jiyong, S., Xu, J.-L., & Sun, D.-W. (2019). Recent progress in rapid analyses of vitamins, phenolic, and volatile compounds in foods using vibrational spectroscopy combined with chemometrics: A review. Food Analytical Methods, 12(10), 2361
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2641. Tahir, H. E., Zou, X. B., Shi, J. Y., Mariod, A. A., & Wiliam, T. (2016). Rapid determination of antioxidant compounds and antioxidant activity of Sudanese Karkade (Hibiscus sabdariffa L.) using near infrared spectroscopy. Food Analytical Methods, 9(5), 1228
1236. Tsai, P.-J., McIntosh, J., Pearce, P., Camden, B., & Jordan, B. R. (2002). Anthocyanin and antioxidant capacity in Roselle (Hibiscus Sabdariffa L.) extract. Food Research International, 35, 351
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543.

CHAPTER

14

Hibiscus sabdariffa extract: antimicrobial prospects in food pathogens and mycotoxins management

Lydia Quansah1, Gustav Komla Mahunu2, Haroon Elrasheid Tahir3, Maurice Tibiru Apaliya4, Mildred Osei-Kwarteng5 and Abdalbasit Adam Mariod6,7 1

Department of Biotechnology, Faculty of Biosciences, University for Development Studies, Tamale, Ghana 2 Department of Food Science and Technology, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana 3 School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China 4 Department of Food Science and Postharvest Technology, Faculty of Applied Sciences, Cape Coast Technical University, Cape Coast, Ghana 5 Department of Horticulture, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana 6 Indigenous Knowledge and Heritage Center, Ghibaish College of Science & Technology, Ghibaish, Sudan 7 College of Sciences and Arts-Alkamil, University of Jeddah, Alkamil, Saudi Arabia

Chapter Outline 14.1 Introduction ...............................................................................................216 14.2 Antioxidant properties in H. sabdariffa .........................................................216 14.3 Antimicrobial potentials of H. sabdariffa ......................................................218 14.4 Mycotoxin production and economic losses to food ......................................220 14.5 Mycotoxin effect on human health ...............................................................220 14.6 Synthetic fungicides: implications on food quality and consumer health ........222 14.7 H. sabdariffa extract: biocontrol agent against pathogens and mycotoxins .....222 14.8 Conclusion .................................................................................................223 References ..........................................................................................................223

Roselle (Hibiscus sabdariffa). DOI: https://doi.org/10.1016/B978-0-12-822100-6.00003-3 © 2021 Elsevier Inc. All rights reserved.

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14.1 Introduction Hibiscus sabdariffa is ordinarily termed Roselle. It comes from the family Malvaceae emanating from countries in the tropics and subtropics and grown and utilized extensively in several countries in the world including Africa. The plant has been utilized for cold and hot beverages and, a popular drink is made from its calyx which is widely known as “Zobo” in Nigeria and “Sobolo” in Ghana (Udo, Ben, & Etuk, 2016). Roselle plant has been utilized in the folk treatment of hypertension, bacteria proliferation restriction, as well as for the treatment of digestive issues, and it is also used as a sedative (Portillo-Torres, Bernardino-Nicanor, & Go´mez-Aldapa, 2019). Due to its versatility, the plant is being investigated for its possible effect against foodborne pathogens and mycotoxins. Foodborne pathogens and mycotoxins are a major problem in developed and developing countries (Yeni, Acar, Polat, Soyer, & Alpas, 2014). Pathogens arising from food can lead to the development of several infections in humans (Adeyeye, 2016). The use of synthetic antibiotics has been the main source for the treatment of bacterial infections arising from foodborne pathogens. However, the continuous overuse, unsupervised use, and abuse of these drugs have contributed to the development of multidrug resistant strains of several microorganisms (Vastrad & Byadgi, 2018). Mycotoxin producing fungi are also a problem in fresh ready-to-eat foods, legume, and cereal production which can arise on the field or during handling, postharvest storage, transportation, and processing (Garcia, Ramos, Sanchis, & Mar´ın, 2009). These mycotoxins are found to induce carcinogenicity, hepatoxicity, teratogenicity, genotoxicity, nephrotoxicity, and are shown to cause reproductive disorders in animal studies (De Boevre, Jacxsens, & Lachat, 2013). Management or prevention of mycotoxin contamination in food is done using synthetic antimicrobials which can be toxic to humans as well as environment as well as generating resistance in the pathogens due to excessive use. Therefore ongoing research is geared toward investigating the potential of plant natural products as an alternative to these synthetic drugs (Vastrad & Byadgi, 2018). Extracts of plants which are obtained after extraction may well be used as therapeutic agents that are usually anticipated to consist of phytochemicals. These compounds are the natural defense structure of plants against diseases and pests (Habib, Aziz, & Karim, 2010). The potential of this versatile plant to be used as an antimicrobial against food pathogens and mycotoxins is reviewed.

14.2 Antioxidant properties in H. sabdariffa Bioactive compounds have been revealed to be abundant in H. sabdariffa plant (Jabeur, Pereira, & Barros, 2017; Lin, Chen, & Wang, 2011). Several accessions of H. sabdariffa have been reported to contain varying concentration in antioxidants (Wang et al., 2014; Wang, Cao, & Ferchaud, 2016). The phenolic content

14.2 Antioxidant properties in H. sabdariffa

in the plant consists mainly of anthocyanin and flavonoids, and it has been shown that the phenolic content contributes the highest to its antioxidant activity (Anokwuru, Esiaba, Ajibaye, & Adesuyi, 2011). Antioxidant compounds and antioxidant activity have been found in the leaves (Wang et al., 2014, 2016; Zhang et al., 2011; Zhen, Qi, & Chin, 2015), calyx (Anokwuru et al., 2011; Hirunpanich, Utaipat, & Morales, 2006; Oboh & Rocha, 2008), seeds (Bako, Mabrouk, & Abubakar, 2019; Mohamed, Ferna´ndez, Pineda, & Aguilar, 2007; Mohd-Esa, Hern, Ismail, & Yee, 2010), and stem. Major antioxidant compounds contained in the leaves of different accessions include isoquercitrin, chlorogenic acid, neocholorogenic acid, cryptochlorogenic acid, and rutin with the latter being the most abundant antioxidant in the leaves (Wang et al., 2014). The antioxidant scavenging activity has been shown for different parts of the plant (Mohd-Esa et al., 2010) with the seed having the highest abundant antioxidant activity. Furthermore, the presence of amino acids mostly in the seed extract and seed oil has been shown to further contribute to the free radical scavenging activity of the plant (Tounkara, Bashari, Le, & Shi, 2014). However, it has been reported that geographical location, soil, and extraction method might highly influence the amount of antioxidants in the various parts (Ochani & D’mello, 2009; Zhen et al., 2015). For instance, phenolic content in the seed, calyx, leaf, and stem was found to be 2.97, 1.85, 1.71, and 0.90 mg of GAE/g, respectively when extracted with water, whereas extraction with methanol yielded 4.87, 2.91, 2.20, and 1.31 mg of GAE/g in the seed, calyx, leaf, and stem, respectively (Mohd-Esa et al., 2010). An herbal concoction made from the leaves and especially the red calyx of H. sabdariffa contains antioxidant properties (Prenesti, Berto, Daniele, & Toso, 2007; Tsai, McIntosh, Pearce, Camden, & Jordan, 2002; Zhen et al., 2015) which has been demonstrated to be effective against generation of reactive oxygen species (ROS) in cells. ROS are generated during normal cellular metabolic processes in humans, but their overproduction can depress the intracellular antioxidant leading to activation of lipid peroxidation, DNA breaks, and protein modification (Stanner, Hughes, Kelly, & Buttriss, 2004). These lead to induction of several diseases including and not limited to cardiovascular diseases (CVD), inflammations, cancer, in humans. ROS are controlled by antioxidant defense and thus a system’s antioxidant defense is the only way to bring ROS production under normal levels. The antioxidant potential of H. sabdariffa has been seen in studies to be effective against diseases such as CVD. Different parts of the plant including the leaves, calyx, stem, and seeds (Mohd-Esa et al., 2010; Ochani & D’mello, 2009; Usoh, Akpan, Etim, & Farombi, 2005) have been found to contain high content of antioxidant. The antioxidant potential has been demonstrated in vivo where rats treated with H. sabdariffa extract showed decreased sodium-induced hepatic peroxidation when compared with the control group (Usoh et al., 2005). Antioxidants are used in the preservation of food to increase the shelf life. Synthetic antioxidants such as butylated hydroxyanisole and butylated hydroxytoluene (Vicetti, Ishitani, Salas, & Ayala, 2005) are mostly used to extend the shelf

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life of food. The excessive use of these synthetic antioxidants as additives in stabilizing food during storage is known to cause health implications (Chan, Khong, Iqbal, Ch’Ng, & Babji, 2012) and thus the interest in using natural alternatives from plant origin. Different parts of H. sabdariffa have been reported to contain high antioxidant content (Anokwuru et al., 2011; Oboh & Rocha, 2008; Ochani & D’mello, 2009). Due to the antioxidant nature of H. sabdariffa, interest for it being used for stabilizing oils and fat has taken precedence. Oil and extract of H. sabdariffa were shown to stabilize antioxidants in sunflower oil compared with using tocopherol (Nyam, Teh, Tan, & Kamariah, 2012) and thus enhanced the shelf life. In a study to extend the shelf life of sardines, it was found that 5% Hibiscus flower inhibited microbial growth, whereas the combination of 5% each of Hibiscus flower and Pecan nut increased the shelf life with no adverse effect on quality indexes (Villasante, Girbal, Meto´n, & Almajano, 2018).

14.3 Antimicrobial potentials of H. sabdariffa Bacteria such as Salmonella typhimurium, Staphylococcus aureus, Escherichia coli, Bacillus cereus, and Listeria monocytogenes and many other microbes are foodborne pathogens that contribute to food spoilage and are of public health concerns (Gizaw, 2019; Vindigni, Srijan, & Wongstitwilairoong, 2007). Methicillin-resistant Staphylococcus aureus (MRSA) is a known healthcare-associated pathogen but has been widely reported to occur in food products especially of animal origin (Feßler, Kadlec, & Hassel, 2011; Normanno, Corrente, & La Salandra, 2007; Vanderhaeghen, Hermans, Haesebrouck, & Butaye, 2010) and is major cause of foodborne illnesses in many countries. Most MRSA isolates from food samples are resistant to a number of commonly used antimicrobials with quite a number of the isolates being reported to be multidrug-resistant (Doyle, Hartmann, & Wong, 2012; Normanno et al., 2007). These foodborne pathogens can cause significant losses in the food chain thus becoming a public health threat when food contaminated with these pathogens are consumed by humans (Gizaw, 2019). Food spoilage pathogens are mostly controlled using synthetic antimicrobials and the excessive usage of antimicrobial has rendered several antimicrobials susceptible to MRSA (Al-Ashmawy, Sallam, Abd-Elghany, Elhadidy, & Tamura, 2016). Furthermore, the use of synthetic antimicrobials in especially livestock production may cause the presence of antimicrobial residues in the food chain and induce harmful reactions when these food products are consumed as well as posing danger to the environment. Thus other alternatives to synthetic antimicrobials are being explored. In recent years, the interest in plant extracts as antimicrobial continues to receive attention and many plants have been found to be of great potential (Lewis & Elvin-Lewis, 1995). One of such plants is the Roselle plant, a versatile plant found to contain several phytochemicals (Da-Costa-Rocha, Bonnlaender, Sievers, Pischel, & Heinrich, 2014) reported to contribute to the antimicrobial potential of the plant (Ali, Al Wabel, & Blunden, 2005; Mahadevan & Kamboj, 2009).

14.3 Antimicrobial potentials of H. sabdariffa

Roselle extracts including leave, calyx, and seed have been shown to possess the ability to inhibit the growth of both gram-negative and gram-positive bacteria isolates from food, veterinary, and clinical samples (Abd-Ulgadir, Suliman, Zakria, El-Deen, & Hassan, 2015; Al-hashimi, 2012; Fullerton, Khatiwada, Johnson, Davis, & Williams, 2011). Besides, the extract has also been reported to inhibit the growth of both susceptible and resistant strains of bacteria (Yin & Chao, 2008). Calyx extract was shown to inhibit E. coli 157:H7 isolated from food, veterinary, and clinical samples with higher inhibition zone occurring for the veterinary samples (Fullerton et al., 2011). In another study, the calyx extract exhibited antimicrobial activity against selected organisms ensued at concentrations with a minimum inhibition zone of 0.30
1.30 mg/mL against S. aureus, Bacillus stearothermophilus, Micrococcus luteus, Serratia mascences, and others (Tolulope, 2007). The leaves have also been shown to possess antimicrobials effect against S. aureus, Salmonella typhi, and E. coli (Sulemana, 2019). The seed also possesses antimicrobial potential (Nwaiwu, Mshelia, & Raufu, 2012). The aqueous extract of the calyx has been investigated for its potential as an antimicrobial in ready-to-eat-food. In one study, the calyx extract used as antimicrobial rinse was found to be effective against L. monocytogenes and MRSA in hot dogs (Higginbotham, Burris, Zivanovic, Michael Davidson, & Neal Stewart, 2014). Furthermore, calyx extract used on fruits and vegetables was also found to be effective to either reduce or totally control foodborne pathogens. For example, calyx extract was effective against 13 foodborne pathogens found in two pepper cultivars in Mexico compared with treatment with sodium hypochlorite, colloidal silver, and acetic acid as rinse (Rangel-Vargas, Go´mez-Aldapa, & Falfan-Cortes, 2017). The dry calyx extract was able to reduce the concentration of 11 foodborne pathogens inoculated on whole and cut mangos (Rangel-Vargas, Luna-Rojo, & Cadena-Ram´ırez, 2018). Not only is the plant effective against bacterial but the antiviral activity has also been studied. Aichi virus (AiV) is a foodborne pathogen that has been shown to cause gastroenteritis which is problematic to control due to the absence of vaccine. It was found that aqueous extracts of H. sabdariffa concentration of 200 and 100 mg/mL were able to reduce AiV to nondetectable levels after 24 hours (D’Souza, Dice, & Davidson, 2016). In another study, calyx extract concentrations of 20, 40, and 60 mg/mL were treated either by autoclaving or by filtration and tested on foodborne pathogens strains of two each of E. coli and S. aureus in a microbiological medium and milk treated with the pathogens. The results indicated that the extract concentration of 40 mg/mL exhibited greater activity in the microbiological medium. For the activity in milk, it was revealed that S. aureus was inactivated in whole and skimmed milk after 168 hours with 40 mg/mL extract, whereas that of E. coli was inactivated after 96 hours with 60 mg/mL extract (Higginbotham, Burris, Zivanovic, Davidson, & Stewart, 2014). The above evidence points to the fact that H. sabdariffa extract can have an important position to be used in the food industry to curb the presence of foodborne pathogens.

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14.4 Mycotoxin production and economic losses to food Food spoilage caused by fungi is a major problem that leads to tremendous economic losses worldwide (Garcia et al., 2009). Furthermore, it can present a health risk to consumers due to potential of fungi to produce mycotoxins (Serrano, Font, Ruiz, & Ferrer, 2012). Mycotoxins are secondary metabolites produced by fungi such as Aspergillus, Penicillium, and Fusarium spp., (Galvano, Piva, Ritieni, & Galvano, 2001). Mycotoxins production in food has become a major problem due to the danger it poses to food safety and health (Alshannaq & Yu, 2017). It is also responsible for food insecurity in tropical regions, especially in Africa (Pitt, Taniwaki, & Cole, 2013). Climatic changes might lead to increase mycotoxin contamination in agricultural commodities and thus it is envisaged that food insecurity issues might increase, especially in Africa. Therefore it is important to find solutions to prevent or bring to a minimum pathogen causing mycotoxins in food and feedstuff. Mycotoxin detection and occurrence in food crops and animal products have been extensively studied (Table 14.1). Mycotoxin in food occurs on the field, during harvesting, handling, storage, and processing (Neme & Mohammed, 2017). Mycotoxin production by fungi has become a major concern as it contaminates plant-based foods and food of animal origin (Markov et al., 2013; Neme & Mohammed, 2017). Mycotoxins such as “aflatoxin, ochratoxins, trichothecenes, zearalenone, fumonisins, tremorgenic toxins, and ergot alkaloids” (Zain, 2011) are of major concerns in agricultural commodity trade with significant economic importance. Mold or fungi growth has been estimated to affect 25% of the world’s crops. Mycotoxin contamination in the food chain affects economies of both developed and developing countries, though the impact is felt more in developing countries due to favorable environmental conditions.

14.5 Mycotoxin effect on human health Several mycotoxins such as Aflatoxin B1, B2, G, ochratoxin A, fumonisin, zearalenone, deoxynivalenol, and T2 toxins are known dangerous toxins that have health implications for consumers when ingested (Serrano et al., 2012). The main toxic effects of exposure to or continuous ingestion of even low levels of mycotoxins are carcinogenicity, hepatoxicity, teratogenicity, genotoxicity, nephrotoxicity, and can cause reproductive disorders (Galvano et al., 2001; Ur Rahman et al., 2020). It may also cause retardation in growth, especially for children under 5 years which is a common occurrence in developing countries due to ingestion of unknown infested food. Humans are exposed to mycotoxin through consumption of contaminated plant-derived foods (Bryden, 2007; Reddy et al., 2010) and sometimes spores of mycotoxins can be present in indoor environment which finds their way through inhalation (Robbins, Swenson, Nealley, Gots, & Kelman, 2000). Animal-based

14.5 Mycotoxin effect on human health

Table 14.1 Detection methods for determining mycotoxins in food and feed samples. Detection method

Product contaminated

Aflatoxin

PCR, LC-MS/ MS, TLC, HPLC, GC-MS, Multiplex lateral flow immunoassay, Molecular imprinting polymers, Reveal Q 1 test strips

Stored wheat grains, maize, cereal and legume blend, poultry products, wheat flour products

Fusarium spp.

Deoxynivalenol, Zearalenone, Fumonisins, Trichothecenes

LC-MS/MS, Multiplex lateral flow Immunoassay

Penicillium spp.

Ochratoxin A

LC-MS/MS, ELISA

Stored wheat, grains, wheat flour, wheat flour products, bread, grains (wheat, rye, barley), corn, feed, soy products, pasta, breakfast snacks Stored wheat grains, corn

Species

Mycotoxin

Aspergillus spp.

References Appell and Mueller (2016), Awad et al. (2019), Di Nardo et al. (2019), Opoku et al. (2018), Rahmani et al. (2009), Sadhasivam et al. (2017), Shephard (2016), Sulyok et al. (2006) Di Nardo et al. (2019), Sadhasivam et al. (2017), Biselli and Hummert (2005)

Sadhasivam et al. (2017), Zheng et al. (2005)

products, such as meat, milk, eggs, may sometimes be contaminated through the carryover of mycotoxins and their metabolites (CAST, 2003). It is revealed that approximately 4.5 billion people living in developing countries are often exposed to unrestrained levels of aflatoxin due to this toxin being commonly present in staple diet (Williams et al., 2004). In Africa, aflatoxin levels have been reported to be high in cereals and legumes due to these crops being susceptible to aflatoxin contamination (Atongbiik Achaglinkame, Opoku, & Amagloh, 2017). It is also reported that aflatoxin is a known human liver carcinogen and is responsible for Hepatocellular carcinoma or liver cancer, which is among the leading cancer deaths worldwide but with higher incidence in developing countries (Liu & Wu, 2010). Aflatoxicosis, a disease arising from ingestion of food contaminated with aflatoxin is a major problem in developing countries. Aflatoxicosis can induce hemorrhagic necrosis of the liver, proliferation of the bile duct, edema, and lethargy (Williams et al., 2004). Aflatoxicosis is the main cause of liver cancer but workers handling contaminated grains have been found

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to have a high risk for lung cancer (Kelly, Eaton, Guengerich, & Coulombe, 1997). Aflatoxicosis is a major problem in Eastern Africa with occasional outbreak in Kenyan, which results from people ingesting aflatoxin-contaminated maize (Kimanya, 2015). A study conducted in Kenyan found that 67.9%, 92.9%, and 50% of maize, millet, and sorghum, respectively were contaminated with aflatoxin and had levels of 0.17
5.3 ppb, and 0.14
6.4 ppb for maize and millet, respectively while that of sorghum exceeded the allowable maximum limit of 10 ppb set by the Kenya Bureau of Standards (Sirma, Ouko, & Murithi, 2015). Aflatoxin levels in cereal and legume blends in Ghana were found to be in the range of 1
1094 ppb for cereal
legume blends and 1
11.7 ppb for cereal-onlyblend which far exceeds the acceptable limit of 20 ppb thus this can have health implications by contributing to malnutrition for the population, especially for children under 5 years (Opoku, Achaglinkame, & Amagloh, 2018).

14.6 Synthetic fungicides: implications on food quality and consumer health Fungicides are used to control diseases in crops caused by fungi. Continuous use of synthetic chemicals has increased the development of resistant strains as well as its effect on consumer health as it generates residues on the produce that are treated with these chemicals (Jacometti, Wratten, & Walter, 2010). It also leaves residues in soil and water bodies and can disrupt the natural ecosystem (Tripathi & Shukla, 2010). Hence other safer alternatives such as plant extracts with minimal impact on human health and environment are being explored (du Plooy, Regnier, & Combrinck, 2009; Romanazzi, Lichter, Gabler, & Smilanick, 2012). The continuous use of fungicide has rendered pathogens such as Botrytis cinerea to be resistant to almost all commonly used fungicides (Droby & Lichter, 2007; Leroux & Walker, 2013). Laboratory and field studies have shown that in some cases the increase use of some fungicides at sublethal doses rather increased mycotoxin production from Fusarium phytopathogens (D’Mello et al., 1998). Synthetic chemical usage has been found to leave residues, especially in fruits and vegetables. A survey carried out in Denmark found that 10% of selected food commodities had residues of pesticides (Juhler, Lauridsen, Christensen, & Hilbert, 1999).

14.7 H. sabdariffa extract: biocontrol agent against pathogens and mycotoxins Plant extracts have been investigated for their possible use against pathogens and mycotoxins (Chomnawang, Surassmo, Wongsariya, & Bunyapraphatsara, 2009; Galvano et al., 2001), and among such plant-base extracts is the use of H. sabdariffa extract (Camelo-Me´ndez et al., 2013; Goussous, Abu el-Samen, & Tahhan, 2010).

References

Synthetic antifungals are used to control fungi and the continuous overuse has led to the development of resistant strains. Thus to control these resistant strains, it has led to increased dosage of antifungal but this also creates problem by increasing toxic residues in food products (da Cruz Cabral, Ferna´ndez Pinto, & Patriarca, 2013), which becomes a health threat to consumers. Natural products such as plant derivatives have been studied for possible use as antimicrobials to control fungi that produces mycotoxins in the food chain with several promising results shown (da Cruz Cabral et al., 2013). The use of Hibiscus calyx extract has been reported to exert inhibitory effect against Aflatoxin B1 produced by Aspergillus strains. Calyx extract inhibited Aspergillus flavus and Aspergillus parasiticus in the range of 91.5%
97.9% and 87.1%
93.3%, respectively which was better than inhibitory effect exerted by Nigella sativa seed extract which was in the range of 47.9%
58.3% and 32%
48% for Aspergillus flavus and Aspergillus parasiticus, respectively (El-Nagerabi, Al-Bahry, Elshafie, & AlHilali, 2012). In ready-to-eat food, Hibiscus flower extract was investigated against inoculated MRSA and L. monocytogenes on hot dogs. It was found that the flower extract concentration of 240 mg/mL was potent to inhibit and or kill these microbes found on hot dogs (Higginbotham et al., 2014).

14.8 Conclusion Leaves, calyx, stem, and seed of H. sabdariffa contain bioactive compounds with antioxidant and antimicrobial properties which are important to be investigated for their effectiveness against foodborne pathogens and mycotoxin producing fungi. Leaf, calyx, and seed extract activity may be suggestive of the presence of broad-spectrum bioactive compounds. Therefore Roselle extract from leaves, calyx, and seeds could be a promising natural antibacterial agent with prospective uses in the agricultural and food industry to be utilized in preharvest crops on the field as well as for postharvest storage of cereals and legumes that are prone to fungi development. As sometimes mycotoxin contamination can arise from processing plants, extracts can also be explored to be used as cleaning agent in processing plants to prevent mycotoxin contamination in these facilities. This will help in reducing food and food products contaminated with mycotoxins to reduce health risks and economic losses.

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215. Available from https://doi.org/10.1089/fpd.2006.0077. Wang, J., Cao, X., Ferchaud, V., et al. (2016). Variations in chemical fingerprints and major flavonoid contents from the leaves of thirty-one accessions of Hibiscus sabdariffa L. Biomedical Chromatography, 30(6), 880
887. Available from https://doi.org/ 10.1002/bmc.3623.

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Wang, J., Cao, X., Jiang, H., Qi, Y., Chin, K., & Yue, Y. (2014). Antioxidant activity of leaf extracts from different Hibiscus sabdariffa accessions and simultaneous determination five major antioxidant compounds by LC-Q-TOF-MS. Molecules (Basel, Switzerland), 19(12), 21226
21238. Williams, J. H., Phillips, T. D., Jolly, P. E., Stiles, J. K., Jolly, C. M., & Aggarwal, D. (2004). Human aflatoxicosis in developing countries: A review of toxicology, exposure, potential health consequences, and interventions. The American Journal of Clinical Nutrition, 80(5), 1106
1122. Available from https://doi.org/10.1093/ajcn/80.5.1106. ¨ . G., Soyer, Y., & Alpas, H. (2014). Rapid and standardized methYeni, F., Acar, S., Polat, O ods for detection of foodborne pathogens and mycotoxins on fresh produce. Food Control, 40(1), 359
367. Available from https://doi.org/10.1016/j.foodcont.2013.12.020. Yin, M. c, & Chao, C. y (2008). Anti-Campylobacter, anti-aerobic, and anti-oxidative effects of Roselle calyx extract and protocatechuic acid in ground beef. International Journal of Food Microbiology, 127(1-2), 73
77. Available from https://doi.org/ 10.1016/j.ijfoodmicro.2008.06.002. Zain, M. E. (2011). Impact of mycotoxins on humans and animals. Journal of Saudi Chemical Society, 15(2), 129
144. Available from https://doi.org/10.1016/j.jscs.2010.06.006. Zhang, M., Hettiarachchy, N. S., Horax, R., Kannan, A., Praisoody, A., & Muhundan, A. (2011). Phytochemicals, antioxidant and antimicrobial activity of Hibiscus sabdariffa, Centella asiatica, Moringa oleifera and Murraya koenigii leaves. Journal of Medicinal Plants Research, 5(30), 6672
6680. Available from https://doi.org/10.5897/ jmpr11.621. Zhen, J., Qi, Y., Chin, K., et al. (2015). Phytochemistry, antioxidant capacity, total phenolic content and anti-inflammatory activity of Hibiscus sabdariffa leaves. Food Chemistry. Available from https://doi.org/10.1016/j.foodchem.2015.06.006. Zheng, Z., Hanneken, J., Houchins, D., King, R. S., Lee, P., & Richard, J. L. (2005). Validation of an ELISA test kit for the detection of ochratoxin A in several food commodities by comparison with HPLC. Mycopathologia, 159(2), 265
272. Available from https://doi.org/10.1007/s11046-004-8663-3.

CHAPTER

Ethnobotanical uses, fermentation studies and indigenous preferences of Hibiscus sabdariffa

15

Haroon Elrasheid Tahir1, Gustav Komla Mahunu2, Zou Xiaobo1 and Abdalbasit Adam Mariod3,4 1

School of Food and Biological Engineering, Jiangsu University, Zhenjiang, P.R. China Department of Food Science and Technology, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana 3 Indigenous Knowledge and Heritage Center, Ghibaish College of Science & Technology, Ghibaish, Sudan 4 College of Sciences and Arts-Alkamil, University of Jeddah, Alkamil, Saudi Arabia 2

Chapter Outline 15.1 Introduction ...............................................................................................231 15.2 Ethnobotanical uses ...................................................................................234 15.3 Fermented studies ......................................................................................234 15.3.1 Seeds fermented products .......................................................234 15.3.2 Roselle calyx’s fermented products ...........................................240 15.3.3 Uses of Roselle in fermented milk ............................................243 15.3.4 Uses of Roselle in baked products ............................................244 15.4 Indigenous preferences ...............................................................................246 15.4.1 Roselle beverage .....................................................................246 15.4.2 Seeds .....................................................................................247 15.4.3 Leaves ...................................................................................248 15.4.4 Fermented milk .......................................................................248 15.5 Conclusion .................................................................................................248 References ..........................................................................................................249

15.1 Introduction Indigenous information on plant species is the consequence of human association and determination of the most attractive, amazing, and fruitful plant species found Roselle (Hibiscus sabdariffa). DOI: https://doi.org/10.1016/B978-0-12-822100-6.00013-6 © 2021 Elsevier Inc. All rights reserved.

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in the prompt climate at a particular time-frame. Indigenous communities are distinct social and cultural groups that share traditional knowledge from a generation to another on plant species utilized for various intentions for example medicine, pesticides, food, beverages, dyes, aromatics, resins, gums, and other purposes (Petrovska, 2012). Such ethnobotanical knowledge in indigenous peoples are at risk of disappearing because the information passed down orally rather than written and is worsen by modernization. Thus, there is an urgent need to document such knowledge. Roselle (Hibiscus sabdariffa, Malvaceae) calyces are used for producing herbal drinks, hot and cold beverages, fermented drinks, wine, jam, jellied confectionaries, ice cream, chocolates, puddings, and cakes (Cid-Ortega & Guerrero-Beltra´n, 2015; Okpara & Ugwuanyi, 2017). Fermented Roselle seeds play an important role in the human diet especially in developing countries like Africa (Abu El Gasim & Mohammed, 2008; Bouba Adji, Mbofung, & Thouvenot, 2007; Ismail, Ikram, & Nazri, 2008). Although Roselle remains one of the most popular and widely utilized as food and medicinal plants as well as the most highly investigated species in the genus Malvaceae their ethnobotanical studies are limited (Table 15.1). Roselle seeds have been used in the preparation of seasoning agents by fermentation (Abu El Gasim & Mohammed, 2008; Compaore´ et al., 2013). The quality of seeds used in preparing seasoning agents might differ from one region to the other even for the production of the same flavoring agent. Also, the production procedures of Roselle indigenous fermented products are very complex and in many cases, only partly understood regardless of many research efforts (Iwuoha & Eke, 1996). Similarly, the name of the seasoning agents varied from one region to other, for example, in Burkina Faso known as Bikalga (Parkouda, Diawara, & Ouoba, 2008), Mari-Bi in Niger (Rabiou, Lewamy, Tchicama, Alma, & Sadou, 2019), Mbuja in Cameroon (Bouba Adji et al., 2007; Mohamadou, Mbofung, & Thouvenot, 2009), dawadawa botso in Nigeria (Ibrahim et al., 2018), Fururndu in Sudan (Abu El Gasim & Mohammed, 2008), and datou in Mali (Tounkara, Amadou, Le, & Shi, 2011). From the above-mentioned information, it is clear that the indigenous fermented Roselle seeds foods are widely produced in African countries and their preparation remains a traditional family art technology, driven by rural women who have less of the science underpinning their practices. Generally, the method used by rural women is time-consuming, laborious, poorly reproducible with poor productivity. Therefore, great efforts are needed for the development of large-scale-good manufacturing practices for producing these condiments to guarantee the safety and quality of the final product. Sensory properties remain a challenge in the path to the market success of food, as several of the Roselle and Roselle-containing product with important health benefits often translate into products with low acceptability (Arslaner, Salik, & Bakirci, 2020; Iwalokun & Shittu, 2007; Monteiro, Costa, Tomlins, & Pintado, 2019); and consumers are always reluctant to compromise on taste for health (Ares, Barreiro, Deliza, Gime´nez, & Ga´mbaro, 2010; Sabbe, Verbeke, Deliza, Matta, & Van Damme, 2009). Nowadays, understanding attitudes and perceptions and the main

Table 15.1 Ethnobotanical uses of Roselle (Hibiscus sabdariffa). Countries

Part used

Thai

Leaves and tender stems Ghana Leaves Senegal Green leaves and green calyx China and West Seeds Africa, Malaysia Malaysia, Leaves Ghana, Côte d’Ivoire India Calyces Thailand Calyces Sudan Seeds India Tender leaves and stem Malaysia Seeds Northern Seeds Nigeria Sudan Seeds Sudan Leaves Senegal Southern India

Leaves Leaves, petals, fleshy calyces

Ethnobotanical uses

References

Eaten raw in a salad or cooked alone or with meat or fish

Riaz and Chopra (2018)

The most popular traditional leafy vegetables on the Tamale market. Entire leaves steamed leaves of Bambara are used in common local dish known as “thiebou dieun”.

Quaye et al. (2009) Diouf et al. (2007)

The seeds produce vegetable oil that is edible for human consumption

Dalziel and Hutchinson (1948), Atta and Imaizumi (2002), Chan and Ismail (2009) Ismail et al. (2008), Mahyao et al. (2008), Ataogye (2012)

The young leaves and tender stems of Roselle are eaten raw in salads or cooked as greens alone or in combination with other vegetables or with meat or fish Used for sauces preparation In Thailand, people consume Roselle juice to quench thirst Cooked with onion or groundnut, eaten green or dry Used for making chutney during the rainy season, eaten as a salad, used as vegetable and women folk dry it for use during the off-season Used to enriched cookies with fiber and antioxidants The seeds are fermented in the addition of some spices for the production of a food item known as Mungza Ntusa. Seeds are fermented to produce a meat substitute condiment In Kordofan and Darfur, the leaves used for preparation of well-known sauce called Mulah or Karaso Used as spinach, leafy vegetable and condiment Used for making pickles and sour recipes which have medicinal benefits.

D’Heureux-Calix and Badrie (2004) Vasavi et al. (2019) Ismail et al. (2008) Singh, Sureja, and Singh (2006) Nyam, Leao, Tan, and Long (2014) Balami (1998) Yagoub et al. (2004) Suliman, Ali, Idriss, and Abdualrahman (2011) Mathieu and Meissa (2007) Abat, Kumar, and Mohanty (2017)

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CHAPTER 15 Ethnobotanical uses, fermentated studies, and preferences

chemical-sensory drivers of acceptance of beverages and condiments as well as formulating new products that effectively satisfy consumers’ preferences are therefore amid the vital challenges faced by the food and beverage processors (Koester & Mojet, 2012; Ko¨ster & Mojet, 2012; Sun-Waterhouse, 2011). The main objective of this chapter was to review the most relevant aspects regarding the ethnobotanical uses, fermentation studies, and indigenous preferences of Hibiscus sabdariffa.

15.2 Ethnobotanical uses Ethnobotany, the basis of beneficial knowledge on Roselle in their relationship with the traditional or local flora uses (Carvajal-Zarrabal et al., 2012). Generally, the term “ethnobotanical uses” describes the utilization of plants for purposes other than pharmaceutical (Ahmad, Ahmad, & Naqvi, 2017). In this chapter the ethnobotanical uses of various parts of Roselle including calyces, seeds, and leaves, were compiled and critically evaluated, focusing on their non-medicinal usage (Diouf, Gueye, Faye, Dleme, & Lo, 2007; Ismail et al., 2008; Qi, Chin, Malekian, Berhane, & Gager, 2005). A summary of the ethnobotanical uses of Roselle plants in various countries is revealed in Table 15.1.

15.3 Fermented studies 15.3.1 Seeds fermented products Several traditional African condiments are produced by alkaline fermentation of Roselle (Hibiscus sabdariffa) seeds. One such is known as Fururndu (Sudan), dawadawa botso or Mari-Bi (Niger), Mbuja (Cameroon), datou (Mali) and Bikalga (Burkina Faso). Parkouda et al. (2008) studied the preparation and physicochemical properties of Bikalga at different production areas in Burkina Faso and constructed the specific steps for production (Fig. 15.1). Preparation of Bikalga includes several stages with cleaning, cooking, fermentation, steaming, and drying being the most important steps. During the process, the change in pH, approximate composition and minerals were measured. The authors observed that cooking and fermentation increased pH value (8
9) while steaming and drying decreased the value (pH 5 6). The composition of alkaline fermentation seeds was 26.47% for total proteins, 23.19% for lipids and 13.7% for total carbohydrates. Besides, the fermentation increased the moisture content (8.24%), total titratable acidity (1.52%), ash (9.03%), total acidity as well as fat acidity (about 89%). Measurement of the mineral content of fermented seeds of Roselle indicated that fermentation led to significant increases in the concentrations of sulfur, sodium, calcium, and potassium in the final product.

15.3 Fermented studies

Roselle seeds raw

Dry cleaned seeds

Cooking (12-24 hr)

Cooked seeds

Covered recipient (Clay or metallic vessel) Fermentation (3-4 days) Crushing or deep pounding

First fermentation (2-3 days)

Deep pounding

Moulding (small balls)

Moulding (big balls)

Steaming (overnight)

2 nd fermentation under mild

Drying

Remoulding

Bikalga

Steaming (overnight) steaming Drying

Bikalga

FIGURE 15.1 Flow diagram of Bikalga production (Parkouda et al., 2008).

Mari-Bi is a traditional seasoning agent prepared from Roselle fermented seeds of certain varieties of Roselle. In Niger, Rabiou et al. (2019) reported the manufacturing procedures and physicochemical analyzes of Mari-Bi (Fig. 15.2A). The results indicated that fermentation is a key stage that affects nutritional and sensory properties. The authors found that the Mari-Bi is a good source of protein, fat, and some minerals including calcium, potassium, magnesium, manganese, sodium, zinc, and iron. Despite its strong flavor and nutritional potential, its commercialization is extremely threatened by industrial condiments. It is thus essential to improve this product by enhancing the hygienic, organoleptic and sensorial qualities.

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(A)

(B)

Roselle seeds raw

Hibiscus sabdariffa raw

Dry cleaned seeds

Dry cleaned seeds

(C) Roselle seeds raw

Dry cleaned seeds Water washing

Cooking at 90-100°C; 50-80 min Cooking at 100°C for 25

Cleaned and selected

Brown cooked seeds Draining of water Spinning and cooking alkaline medium seeds (17-47 mn; 90-100°C) Sun drying for 24 h

Cooking (8-10h)

Cooked seeds

Black cooked seeds First fermentation (2 days) 1 st fermentation (48 h or 72 h); milling and 2nd fermentation (24h)

Milling Deep pounding

Fermented seeds paste

Uncontrolled fermentation for 9 days Mixing with ash leacheate

Steaming and deodorizing for 40 mn

Sun drying for 24 h

2and fermentation

Deodorized seeds paste Milling

Drying under sun for 2-3 days

Drying for one to several days Dawadawan botso’ Fururndu Mari-Bi

FIGURE 15.2 Diagram of traditional process of production of Mari-Bi (A), Fururndu (B) and Dawadawan botso’ (C) (Ibrahim, Sani, Aliero et al., 2011; Rabiou et al., 2019).

In western Sudan, the art of food fermentation is widespread and essential to the food security of communities. Apart from fermented foods, the seeds of Roselle are cooked and fermented to produce a traditional fermented condiment “Fururndu” that is used to enhance the organoleptic properties of various foods. Fururndu production includes several stages with cooking and fermentations being the most important steps (Fig. 15.2B) (Abu El Gasim & Mohammed, 2008). It is prepared by a solid-state fermentation procedure. It is classified as one of the strong-smelling West African fermented condiments (Dirar, 1993). The flavor is deemed the quality indicator of indigenous fermented condiments and has a vital role in consumer preferences (Ugwuanyi, 2016). In 1993, Dirar has reviewed meat substitutes of plant origin in Sudan processors and he indicated that fururndu is characterized by flavor similar to that of fermented meat to some extent. Yagoub, Mohamed, Ahmed, and El Tinay (2004) evaluated the changes in the functional properties of raw and cooked Roselle seeds due to fururndu fermentation. Dried Roselle seeds were cooked and then fermented for nine days. In vitro

15.3 Fermented studies

digestibility of the seed proteins reached the maximum value (82.7%) after six days of fermentation, then significantly decreased (68.92%). The cooking process significantly (P , 0.01) reduced the in vitro digestibility of the Roselle seed to 69.84%. In another study, Omer and Yagoub (2007) investigated the chemical composition, anti-nutritional factors, HCl-extractability of minerals, amino acid profile, in vitro protein digestibility and microbial growth of raw and sprouted Roselle seeds. The results showed for both sprouts (24 or 48 hours) reduced total acidity, fat acidity and, pH indication leaching of acids during soaking before sprouting. During the fermentation process of sprouted seed, the total acidity increase was accompanied by an increase in the pH to 6.00 followed by a decrease to 5.79 for 24 and 48 hours sprouted seeds, respectively. Fururndu made from sprouted seeds increased total polyphenols while phytic acid remains constant. The increase of polyphenols could be attributed to the release of bound polyphenols resulting from microbial activity. Fermentation of sprouted seed not only has a positive effect on polyphenols but also a beneficial effect on the bioavailability of Ca, Mn, and Fe. Fermented sprouted seeds, on the other hand, has a negative effect on potassium and sodium. In vitro digestibility of the raw seed proteins were 51.465%, and fermentation of both sprouts (24 and 48 hours) considerably reduced it to 31.27 and 28.70% respectively. During fururndu preparation, the digestibility of the two sprouts (24 and 48 hours) was respectively increased to values of 48.293% and 48.780%. Dawadawa botso is a traditional condiment and is widely consumed in West Africa. It has gained little scientific attention (Ibrahim et al., 2018). The preparation of dawadawa botso has many steps similar to other fermented Roselle seeds with some variations according to the skills and knowledge of the producers (Ibrahim, Sani, Aliero, & Shinkafi, 2011). Fig. 15.2C shows the flow chart for dawadawa botso preparation. A study conducted on fermented Roselle seeds to produce dawadawa botso in Zuru (Nigeria), showed that it involves high microbial numbers with Bacillus species being the dominant microflora involved in the fermentation (Ibrahim et al., 2018; Ibrahim, Sani, & Shinkafi, 2011). It has been reported that the approximate composition of traditional fermented Roselle seeds was 25.19% for protein, 17.17% for lipid and 15.98% for carbohydrate (Ibrahim et al., 2011). This affirms that dawadawa botso is a good and low-cost source of protein especially for developing countries where the expensive sources of protein are not affordable and its consumption might promote health due to the existence of probiotic bacteria. The major minerals are potassium (13733.33 mg/kg) and sodium (155.83 mg/kg) while the lowest values were observed in magnesium (0.26 mg/kg) and calcium (0.04 mg/kg). The higher concentrations of potassium and sodium in traditionally fermented seeds could be due to the addition of ash leachate during the production process. The composition of dawadawa botso prepared by combinations of fermenting bacteria was found to be within the range of 2.17%
15.50% for lipid, 15.12%
27.56% for protein and 11.04%
40.72% for carbohydrate (Ibrahim et al., 2018). The reported microorganism related to the amino acid profile of dawadawa botso

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was a combination of a variety of Bacillus species such as Bacillus Pumilus, Bacillus subtilis, Bacillus laterosporus, Bacillus polymyxa, Bacillus amyloliquefaciens, Bacillus licheniformis, and Bacillus brevis. Dawadawa botso produced using the mixture of above-mentioned microorganisms exhibited an increase in monosodium glutamate-like amino acid (aspartic acid, and glutamic acid) while sweet (glycine, alanine, serine and threonine) and bitter (such as arginine, leucine and isoleucine) amino acids showed a decrease for the starter combinations used. The authors concluded that the dawadawa botso produced by Pediococcus pentasaceus, Leuconostoc mesenteroides and Lactobacillus plantarum was superior to that fermented with all Bacillus sp. The sodium salt of glutamic acid is the main component in modern seasoning (Steinkraus, 1996). An important point to consider during the Roselle seeds fermentation is the influence of microbial species on the amino acid and volatile compounds profiles of the final products. During dawadawa botso production, the authors found that amino acids and volatile compounds profiles can differ significantly according to the microorganisms used to drive the process (Ibrahim et al., 2018; Ibrahim, Sani, Aliero et al., 2011). In another study, Ibrahim, Sani, Aliero et al. (2011) investigated the main volatile compounds accountable for its unique flavors of fresh and locally produced dried dawadawa botso. A total of 22 volatile compounds were detected in both types of dawadawa botso counting alcohols (3), acids (8), esters (7), aldehydes (3), and alkanes (1). Methyl (9Z)
12- hydroxyl-9
octadecenoate being the major compounds in fresh while methyl (14E)
14,17-octadecadienoate in dried and Cis-9hexedecenal (19.96%, 15.13%) in both types. The old cheesy odor of dawadawa botso can be attributed to the ethyl methyl acetic acid and isobutyl acetic acid. Hexadecanoic acid, methyl ester are accountable for the sweet aroma perceived in dawadawa botso (Ibrahim, Sani, Aliero et al., 2011). Mbuja is a traditional food condiment prepared by fermentation of cooked Roselle seeds in Cameroon. The traditional method production steps are shown in Fig. 15.3. Mohammadou, Mbofung, Mounier, and Coton (2018) investigated the physicochemical properties of Mbuja fermented with different starter cultures. During the fermentation process, the combination of Bacillus amyloliquefaciens (JQ410767), Bacillus amyloliquefaciens (JQ410771), and Bacillus subtilis (JQ410778) were used. As a result of fermentation, proteins, ash, and fats increased while carbohydrates, crude fibers decreased. Bioactive components (such as phenols, flavonoids and carotenoids) with a prospective positive effect on human health were detected and increased by the fermentation. Similarly, Mohamadou, Mbofung, and Thouvenot (2007) reported the bacillus strains in the fermenting seeds of Roselle seeds during Mbuja production can provide both antioxidants and probiotics organisms. When Roselle seeds were fermented for Mbuja production, increases in the concentration of mineral content namely magnesium, sodium, aluminum, iron, manganese, and zinc were noted (Mohamadou, Mbofung, & Thouvenot, 2008). Conversely, the concentration of calcium declined while copper content did not change. Lactic acid bacteria strains have been shown to have a greater contribution to increasing mineral content during the production of Mbuja.

15.3 Fermented studies

Roselle seeds raw Dry cleaned seeds Cooking (>3 100°C)

Draining of water Fermentation in a close earthenware pot for 7 days

Phase 1 of fermentation

Pounding

Fermentation in earthenware pot for 3days

Phase 2 of fermentation

Darying Mbuja

FIGURE 15.3 Flow diagram of Mbuja production (Mohamadou et al., 2009).

In Benin, Yanyanku and Ikpiru are prepared by the fermentation of Roselle seeds and used as functional additives for African locust bean (Parkia biglobosa) seed fermented food condiments which used to improve the flavor of various dishes including soups and sauces, due to their sensory properties and high nutritional value (Agbobatinkpo et al., 2011, 2013; Azokpota, Hounhouigan, & Nago, 2006). Processing of Roselle seeds into Yanyanku and Ikpiru form Roselle includes cooking, draining, alkaline fermentation, pounding/crushing, molding and sun drying (Fig. 15.4). However, differences are found between and within these products according to the skill of the processors. Generally, the pH of the final product is beyond 8 (Agbobatinkpo et al., 2013). In 2013, Agbobatinkpo et al. investigated the biodiversity of aerobic endospore-forming bacterial species occurring in Yanyanku and Ikpiru, fermented seeds of Roselle used to prepare food condiments in Benin (Agbobatinkpo et al., 2013). In this study, the aerobic endospore-forming bacteria (AEFB) resulting from Yanyanku and Ikpiru fermentation were examined by a combination of geno- and phenotypic approaches. Similar AEFB species were identified in both Yanyanku and Ikpiru and mainly consisted of Bacillus spp., with 54% of the isolates were belonging to the Bacillus subtilis group (Bacillus subtilis, Bacillus licheniformis and Bacillus

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FIGURE 15.4 (A) Flow diagram of Ikpiru processing; (B) flow diagram of Yanyanku processing (Agbobatinkpo et al., 2011, 2013).

amyloliquefaciens). Pathogenic spore-forming bacteria were also detected, assuring the need to produce starter cultures for the two functional additives able to inhibit the growth pathogenic bacteria and thus improving the safety and quality of fermented condiments.

15.3.2 Roselle calyx’s fermented products According to previous studies of health benefit of Roselle and consumer interest in functional food that can reduce the risk of cardiovascular diseases, such drinks and fermented beverages have been produced from Roselle calyces and some of them are currently being marketed in the US and Europe (Ifie, Marshall, Ho, & Williamson, 2016; Mounigan & Badrie, 2007). A previous study has reported that the red Roselle flower is rich in anthocyanin and contains various organic acids (Ifie et al., 2016; Shruthi et al., 2016). The anthocyanin of Roselle is stable and vary in color from pink to blue and violet (Shruthi et al., 2016; Yokotsuka, Yajima, Seki, & Matthews, 1997). These properties suggest that the extracts of Roselle calyces can be used as a suitable material for the making of colored wines. The main steps of Roselle wine production involve pasteurization, preparation of Roselle must, yeast starter, and fermentation. The detailed procedures of production and also the length of fermentation of Roselle wine may differ from one maker to another. Mounigan and Badrie (2007) investigated the effects of pretreatments of pectolase and temperature/time on the properties of red Roselle wine. The Roselle calyces were pasteurized at 60 C for 3.5 hours or 90 C for 30 minutes at 0%, 0.5% and 1.0% wt./wt. pectolase addition in fermentation of

15.3 Fermented studies

wines. The authors noted significant changes in color, physiochemical and sensory properties. When compared with 60 C for 3.5 hours, samples treated with 90 C for 30 minutes had pH 2.57, titratable acidity 0.43% as citric acid, total soluble solids (TSS) 10.53 Brix and 15.29% alcohol. The authors attributed the changes in physicochemical parameters to pectolase enzyme while temperature and time had no effects. Alobo and Offonry (2009) studied the characteristics of colored wine produced from Roselle extract. In this study, Saccharomyces cerevisiae has been used as a starter culture. Both must and wine were subjected to physicochemical analysis. The results indicated that ameliorated Roselle must have a pH value of 3.72, 4.21% protein, 0.69% titratable acidity, 21 Brix TSS, ascorbic acid 1.06 mg/ 100 mL, and 28.30 abs/mL anthocyanin. The colored wine had a pH value of 3.43, 1.75% protein, 0.69% titratable acidity, 4.90 Brix TSS, ascorbic acid 0.6 mg/100 mL, 22.65 abs/mL anthocyanin and 10.80% alcohol after aging. The alcohol content of Roselle in this study is less than that reported by Mounigan and Badrie (2007), however, this value is comparable with that reported in lemon wine (10.1%) and a combination of pawpaw and Roselle extracts (10.5%) (Alobo, 2002; Okoro, 2007). The alcohol content of the produced wine is within the standard specification for wines (Amerine et al., 1980). When compared with imported red wine, Roselle wine exhibited no significant difference and it seems that Roselle calyx may be used to produce acceptable colored wine. During fermentation of cocoa juice and Roselle extracts into a wine-like alcoholic beverage, Darman, Ngang, and Etoa (2011) reported deep reddish color due to Roselle extract, higher acidity (8.025 g tartaric/100 ml) and high ethanol content (11.02%). The mean value of phenols was 962 mg/L. The fermentation of cocoa juice and Roselle extract for the production of red wine showed many volatile compounds including alcohol, acids and esters which are important in the sensory properties and quality of wines (Maicas, Gil, Pardo, & Ferrer, 1999). According to this study, isovaleric acid with heavy cheese flavor was found in high concentrations when compared with the other fruit-based commercial wines. A similar result was found in red wines (Gil, Cabellos, Arroyo, & Prodanov, 2006). The authors recorded higher concentration of iron (1.24 mg/L) and lower concentration of Cu (0.69
0.93 mg/L), and Ca (6.0
7.5 mg/L). The higher iron in cocoaRoselle wine can be attributed to Roselle extracts. Ifie et al. (2018) studied the effect of ageing temperatures (6 C, 15 C and  30 C) on the color, phytochemical and bioactivity of Roselle wine over 12 months. At the end of storage, wines kept at 6 C showed the highest color density and lowest polymeric anthocyanins. During ageing, a decrease in the initial concentration of individual phenolic compounds and monomeric anthocyanins leading to the development of anthocyanin-derivatives (pyranoanthocyanins), several compounds were reported for the first time in Roselle wine. At the end of ageing period, Roselle wine volatiles were investigated. In this study, the only identified compounds influenced by ageing temperature were diethyl succinate which increased with higher ageing temperature. This finding is in agreement with the

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study of Garde-Cerda´n, Marselle´s-Fontanet, Arias-Gil, Anc´ın-Azpilicueta, & Mart´ın-Belloso (2008), who found wine aged at 23 C had a higher concentration of diethyl succinate than their counterparts aged at 5 C. According to this study, the processing of Roselle calyces into wine represents a promising alternative to expanding the functional characteristics of this product. The authors suggested further studies on the effect of the raw Roselle (fresh versus dried), oak ageing, and microoxygenation on parameters relating to the quality wine including phenolic and volatile profiles and health-promoting effects (Ifie et al., 2018). Fresh and dried calyces of Roselle are used in the production of many products such as hot and cold drinks and fermented beverages (Da-Costa-Rocha, Bonnlaender, Sievers, Pischel, & Heinrich, 2014). Zobo is a traditional drink produced from the dried calyx of Roselle in West African countries (Omemu, Edema, Atayese, & Obadina, 2006; Yeboah-Awudzi, 2017). Nwafor and Akpomie (2014) investigated the effect of fermentation time on quality parameters of Zobo drink produced from Roselle calyces. Saccharomyces cerevisiae and Aspergillus niger have been used as a starter culture for fermentation for a period of 48 hours whereas measurement for the various parameters was conducted at 12-hour intervals. The treatment had significant differences in the quality parameters such as pH value, titratable acidity, protein, carbohydrate, and vitamin C contents as compared with the fresh product. Fermentation using Saccharomyces cerevisiae produced drink which recorded a higher score for aroma, visual appearance, taste, and overall acceptability than that produced by Aspergillus niger. Acceptability decreased from the 24th hour. From this study, the authors suggested that fermentation for 12 hours with Saccharomyces cerevisiae could be used as an optimum approach for Zobo production as it will decrease waste as a result of inadequate fermentation. Ojokoh, Adetuyi, Akinyosoye, and Oyetayo (2002) investigated the effect of addition of neutralizer (trona) on the fermentation of Roselle calyces. Trona is a hydrated sesquicarbonate of sodium and it is highly basic (Makanjuola & JG, 1973; Nielsen, 1999). The addition of trona to the Roselle calyces increased the initial pH values from 3.3 to 5.3 and the total titratable acid from 0.027% to 0.043%. Several important microorganisms of Roselle fermentation have been isolated and identified including Saccharomyces cerevisiae, Aspergillus niger and Aspergillus flavus. Regarding aerobic bacteria, only Escherichia coli, Bacillus subtilis and Klebsiella sp. were isolated. The chemical composition analysis of fermented samples showed that there was an increase in the content of protein (6.56%), lipid (4.25%), and carbohydrate contents (74.33%) in the fermented sample. The fermented calyces were also containing low concentrations of minerals including Zn (31.86 ppm), Fe (12.35 ppm), Mg (132.00 ppm), Ca (294.31 ppm), Na (214.62 ppm) and K (382.13 ppm). The author concluded that the reveals that trona can be used to neutralize the acid in Roselle calyx and that fermentation can significantly impact the nutritional composition positively. The same authors investigated the effect of different concentrations of trona on the microbial population. They showed that the microbial

15.3 Fermented studies

population increased proportionally with the concentration of trona, particularly for the bacterial population which ranged from 5.18 to 5.59 log10 c.f.u./mL (Ojokoh, 2010). In this study, the alteration in total titratable acidity level was inconsistent. The addition of trona during the fermentation process improved the protein content of the samples but significantly reduced the content of phytic acid and tannin (Ojokoh, 2010; Ojokoh, Adetuyi, & Akinyosoye, 2005).

15.3.3 Uses of Roselle in fermented milk Several studies on the effect of Roselle calyces on the quality of fermented food have been reported (Cid-Ortega & Guerrero-Beltra´n, 2015). Iwalokun and Shittu (2007) assessed the influence of addition (prior and after fermentation) of Roselle calyces extracts (20 mL/L of yogurt) on pH value, total titratable acidity, syneresis, ash, and moisture in low-fat yogurt. The results showed that the addition of calyces’ extract before fermentation, reduced the time of fermentation around 0.7 hours than that of the plain yogurt, indicating that the Roselle extract interacts with either fermentation constituent or fermentation products to hasten curd formation. Furthermore, the synergetic effect of organic acids of Roselle and the starter cultures could speed the pH reduction and clotting of proteins. Syneresis index (0.70 6 0.03 mL/125 g) of this yogurt was substantially less than that of control (yogurt with strawberries) or yogurt added with Roselle extract after fermentation. Yogurt added with Roselle extract was less accepted as compared with the control, perhaps due to the high acidity of the extract that produced an unpleasant taste. Arslaner et al. (2020) studied the effect of adding Roselle flowers marmalade 15% and 20% on the dry matter, ash, protein, pH, titratable acidity, water-soluble dry matter and water activity (aw). Yogurt was made with fresh cow’s milk supplemented with skim milk powder to raise the dry matter to 16%. Also, starch (1%) and pectin (1%) were added to give consistency and then pasteurized at 90 C for 15 minutes. The authors observed that the addition of Roselle marmalade did not indicate a negative influence on the physicochemical, microbiological and sensory characteristics of the yogurt samples. Furthermore, adding Roselle to yogurt samples increased the antioxidant activity, polyphenol and minerals (namely, Fe, Mn, B, and Ba). The authors concluded that the addition of yogurt can be used for producing functional yogurt due to its pleasant and characteristic taste. Iwalokun and Shittu (2007) evaluated the effect of the addition of Roselle extract (20 mL/L of yogurt) on pH, titratable acidity, syneresis, ash, moisture content and organoleptic properties of yogurt. Yogurt was produced from reconstituted milk and mixed with sugar (30 g/L). The studies indicated that the addition of the Roselle, prior fermentation, reduced the time of fermentation by approximately 42 minutes. This could be attributed to the synergistic effects of organic acids in the Roselle extracts and the starter cultures which led to rapid reduction of the pH and clotting of proteins. Thus, syneresis index of this yogurt was substantially lower (0.70 6 0.03 mL/125 g sample) as compared with control

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(yogurt with strawberries) or yogurt mixed with extract after fermentation. Fermented milk was prepared from reconstituted skimmed milk enriched with various concentrations of Roselle extracts (0.1%, 0.2% and 0.3%). Milk was pasteurized at 95 C for 10 minutes, then quickly cooled to 42 C and inoculated with milk starter cultures (0.2 g/L) comprising Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus and Bifidobacterium adolescentis. The inoculated mixture was incubated at 43 C until the pH reached 4.5
4.6. Fermented milk without the addition of extract (0.2%) was used as control. They found that the addition of extract might enhance titratable acidity and pH, bacteria counts, syneresis index, water holding capacity and sensory acceptance of the products (Su, Li, Yang, Hou, & Ye, 2018). Su et al. (2018) evaluated the physicochemical properties of fermented milk containing different concentration [0%, 0.1%, 0.2% and 0.3% (wt./wt.)] of Roselle extracts. Titratable acidity (TA) and pH, bacteria counts, syneresis value, water-holding capacity, and their sensory properties were assessed subsequently. The authors concluded that the addition of 0.2% of Roselle extract could improve the quality of fermented milk and promote hypoglycemic and hypolipidemic activities. Furthermore, the good sensory properties of Roselle extract added fermented milk indicated the possibility of producing the fermented milk formulated in the present study on a commercial scale.

15.3.4 Uses of Roselle in baked products Roselle calyces powder have been used in wheat bread due to their content of fiber, calcium, iron and antioxidant capacities. Mata-Ram´ırez, Serna-Sald´ıvar, Villela-Castrejo´n, Villasen˜or-Dura´n, & Buitimea-Cantu´a (2018) evaluated the effect of powdered Roselle calyces added to wheat flour bread to enrich with fiber, calcium, iron and antioxidant substances. The authors added various concentrations of Roselle powder (3%, 6%, or 9%) then found that a high concentration of Roselle powder increased the dietary fiber, red color and anthocyanin content. Ferulic acid was the major phenolic compound identified and it was an inbound form (97%
98%). The antioxidant content of bread enriched with 9% Roselle powder was 2.7 times higher as compared to the control bread. However, wheat bread enriched 9% Roselle powder had higher hardness (4.79 times) and less sensory properties when compared with the control. A high concentration of Roselle powder impeded yeast-gassing power measured in pressure meters possible due to its acidic pH. Roselle powder added wheat bread showed lower bread height, volume and higher density compared to the control. The authors concluded that the Roselle powder could be used to increase dietary fiber, phytochemicals and antioxidant activity of wheat flour bread. Hulu-mur is one of the most well-known nonalcoholic beverage prepared in Sudan from thin flakes of a fermented combination of un-malted and malted sorghum flour, with added spices (Agab, 1985; Dirar, 1993). It is prepared for special occasions of the holy month of Ramadan (i.e. fasting of the holy lunar month

15.3 Fermented studies

of the Muslims) (Dirar, 1993; Sulieman et al., 2009). Preparation of Hulu-mur including several steps with the preparation of malt (zurrria), drying of the malt, milling of malt, preparation of the dough, preparation of Hulu-mur flavors, fermentation and preparation of baked thin flakes being the most important steps. Feterita sorghum is used as the main source of malt and the flour required for hulu-mur preparation. The best hulu-mur is obtained when an equal amount of malt and grain are used. The malt and flour are separately milled into a fine powder. The flour of grain is cooked into porridge and liquefied. At this stage, the malt flour is added to hot porridge and then the two ingredients thoroughly mixed well and allowed to ferment by the natural flora of the malt in a warm place or under the sun. While fermentation occurs, the selected spices (e.g. cinnamon, ginger, galangal, black pepper, cumin, and coriander) used in hulu-mur ground are ground into powder (Baidab, Hamad, Ahmed, & Ahmed, 2016). Besides, the above spices, the aqueous extract of tamarind, red Roselle water, and dates slurry may be added (Dirar, 1993). The liquid additive and the species powder are mixed with the batter after fermentation proceeds 12 hours. After thorough mixing, the whole mixture is further fermented so that the total fermentation time is 24
36 hours. At this stage, the batter is ready for baking and it is sour, red-brown color, slightly sweet, with strong flavor of malts and species (Dirar, 1993). Prior to baking process, the fermented dough is diluted with water to a batter constancy, spread on a hot ceramic pan, and baked into sheets, which are then dried in the shade for approximately two days (Ibnouf, 2012). For the preparation of drinks, the prepared sheets are dispersed in water and allowed to infuse for 1
2 hours, and the dark reddish-brown supernatant is decanted, and sweetened with sugar (Agab, 1985; Baidab et al., 2016; Ibnouf, 2012). Abdalbasit et al. (2017) reported that the chemical composition of millet flour and millet-based fermented products were significantly affected by the fermentation and preparation method; they reported that the protein content was significantly increased as a result of fermentation, whereas oil and carbohydrates were decreased. In addition to these, fermentation and method of preparation increased some amino acids such as threonine, valine, isoleucine, and histidine. Conversely, aspartic acid, serine, glycine, alanine, tyrosine, phenylalanine, lysine and arginine revealed a decrease value. Similar results were observed by Popoola, Jolaoso, and Akintokun (2005), who stated an increase in values for some of the amino acids in the fermented Cirsium altissimum seeds. Mahgoub, Ahmed, Ahmed, and El Nazeer (1999) evaluated the influence of traditional Sudanese processing of hulu-mur drink on the thiamine, riboflavin and mineral contents. The authors showed that fermentation and baking of hulu-mur can reduce the content of thiamine and riboflavin. The addition of spices to hulu-mur dough resulted in increases in strontium, calcium and iron. The authors concluded that the baking did not cause substantial loss in the contents of minerals. In a study, Baidab et al. (2016) reported significant amount of Na (26.45 mg/100 g), K (21.84 mg/100 g), Ca (24.00 mg/100 g), and Fe (0.57 mg/100 g) in Hulu-mur carbonated beverage analog as compared to concentrations of the same minerals in the nonalcoholic beverage (22.31, 8.19, 22.00 and 0.15 mg/100 g, respectively).

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15.4 Indigenous preferences Roselle and Roselle containing product preferences were recognized as playing a vital role in product choices and consumption. Understanding sensor properties are essential in formulating new products that effectively meet consumers’ expectations and preferences, and are therefore amid the main challenges faced by the food processor nowadays (Monteiro et al., 2019; Sun-Waterhouse, 2011). The choice of Roselle products depends greatly on local consumer preference, while preparation parameters are specified by traditional practices and available technology. Different countries have different preferences for Roselle products.

15.4.1 Roselle beverage Deep red color, aroma, balanced acid, and sweet tastes are the key factors of consumer preference of Roselle beverages (Bechoff et al., 2014; Monteiro et al., 2019). In a study of Alobo and Offonry, panelists of 20 who were familiar with wine were employed to evaluate the sensory properties of Nigerian Roselle wine (Alobo & Offonry, 2009). In this study, the commercial red wine (Baron de Valls) was used as a control. The results indicated there were no substantial differences in color, clarity, flavor, aroma, taste, and overall acceptability between the Roselle wine and the commercial wine. Similar results were found when Ifie, Olurin, and Aina (2012) employed a 20-membered panel to evaluate the sensory quality of the aged Roselle wine in terms of color, flavor, taste, and overall acceptability. From these studies, it can be concluded that acceptable red wine of high quality can be manufactured from Roselle calyces and its industrial perspective should be explored. Mounigan and Badrie (2006) investigated the sensory acceptance of Roselle wines prepared from various calyx puree and total soluble solids. Wine samples were of wt./vol. 20% puree/20 Brix, 30% puree/26 Brix and 30% puree/30 Brix. Roselle wine of 30% puree/26 Brix indicated the highest scores of color, flavor, balance, overall acceptability, and weak in bitterness while the higher puree in Roselle wines led to higher flavor scores, but clarity as percentage emission was lower in wine with 20% Roselle puree. In a study, consumers were asked to evaluate the quality of Nigerian fermented juice prepared from Roselle calyces (Nwafor & Akpomie, 2014). The score of aroma (8.4), visual appearance (8.6), taste (8.0) and overall acceptability (8.8) was achieved when the juice fermented for 12 hours using Saccharomyces cerevisiae as a starter culture. Anderson and Badrie (2015) investigated the sensory acceptability of fermented Roselle. The concentration of calyx to water 20:10 wt./vol. was used in wine preparation. The results showed that enrichment of wine with 25 ppm polyphenol resulted in no significant changes in color and aroma as compared with the control samples but was slightly more astringent. Color and clarity were liked strongly and overall acceptability was moderate. In this study, the awareness of polyphenol and

15.4 Indigenous preferences

purchase intent were also reported. Only 42% of assessors had heard of polyphenols, 24% who knew of the health benefits, 12% linked polyphenols with prevention of cardiovascular diseases and 12% as antioxidants. More than half of assessors liked the enriched Roselle wine and will consume now and then. The authors suggested that providing the consumers with specific benefit besides using exposure as a means to familiarize consumers with sensory characteristics might have a substantial influence on consumer preferences for the functional beverage. In another study, Idolo and Marshall (2019) evaluated the effect of ageing temperature (6 C, 15 C, and 30 C) during storage of 12 months on the sensorial properties of Roselle wine. A panel of 60 was employed for the sensorial analysis. The authors found that Roselle wines aged at 6 C and 15 C were generally ranked better in color, aroma and bouquet, and overall acceptability compared to wine stored at 30 C. The obtained data results will provide Roselle winemakers with the information on postfermentation handling of the wines and provide them a new analytical procedure to attain the desired quality of the final product. In Sudan, the traditional carbonated beverage (i.e. hulu-mur analog) and commercially carbonated beverages (including apple flavor, orange flavor, lemon flavor, and cola flavor) were tested by 15 semi-trained assessors (Baidab et al., 2016). The panelists were asked to assess the differences among the beverages in terms of color, odor, taste, and overall acceptability. The results indicated that the hulu-mur had a comparable quality to lemon flavored carbonated beverage which was already established in the market. Therefore, organoleptically the hulu-mur analog carbonated beverage might compete with other commercial carbonated beverages.

15.4.2 Seeds In Cameroon, a panel of 80 members assessed the taste, texture, flavor, and overall acceptability of Mbuja using various starter cultures (Mohammadou et al., 2018). From the organoleptic viewpoint, Mbuja fermented with the combination Bacillus amyloliquefaciens (JQ410771) and Bacillus subtilis (JQ410778) was the most liked by the hedonic panel, the higher appreciation being driven by the taste and flavor of the product which was quite similar to the reference Mbuja. The same authors investigated the organoleptic properties of Mbuja samples collected from four locations in Cameroon (Dzbam, Gouzda, Magoumaz and Midirey) (Mohamadou et al., 2009). A panel of 13 members was asked to evaluate the samples for taste, color, flavor, and preference of the products. Mbuja collected from Dzbam showed higher sensorial quality while Magoumaz showed inferior quality. No significant differences were observed from samples collected from other locations. The differences in products could be due the variations in the metabolic activity of the various bacteria species which produce different sensory profile for each sample.

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15.4.3 Leaves Quaye, Gyasi, Larweh, Johnson, and Obeng-Aseidu (2009) carried a survey at two main marketing centers in Ghana, namely Tamale and Kumasi. A panel of 100 traders was questioned using a semi-structured questionnaire and focus group discussions held on traders’ perceptions and consumer preferences, relative importance, and indigenous nutritional knowledge of leafy vegetables. The authors found that the order of preference in Kumasi is Xanthosoma mafafa . Corchorus spp.. Amaranthus spp . Hibiscus sabdariffa, while the reverse order pertains in Tamale.

15.4.4 Fermented milk Noviatri, Setianingrum, and Haskito (2020) evaluated the organoleptic properties of yogurt mixed with various concentrations (0%, 0.5%, 1%, and 1.5% vol./vol.) of purple Roselle extracts. The yogurt is made from raw cow’s milk, heated at 72 C for 5 minutes. After cooling to 43 C, the milk was inoculated with an aliquot of starter culture (3%) and incubated at 43 C. A panel of 30 members was asked to evaluate color, taste, aroma, texture and the overall value of yogurt samples. The authors observed that the highest concentration was the most preferable (average score of 3.83) for fortification due it its darker color. In a study, Iwalokun and Shittu (2007) found that the additional Roselle extracts to yogurt have a negative impact on the acceptability of Roselle
fortified yogurt, perhaps due to the excessive acidity of Roselle extract that produced an unpleasant taste. Rasdhari, Parekh, Dave, Patel, and Subhash (2008) adopted the procedure of Iwalokun and Shittu (2007) for yogurt production using pasteurized milk containing probiotics (Lactobacillus casei). Yogurt samples have been fortified with two concentrations (1% and 2% vol./vol.). Generally, the fortification with Roselle extract improved the antioxidant characteristics and decreased syneresis. In contrast to the study of Iwalokun and Shittu (2007), the yogurt sensory acceptability revealed higher scores. The authors suggested that adding Roselle extract to yogurt could improve the quality of yogurt

15.5 Conclusion In this chapter, most of the previous work on ethnobotanical uses, fermentation studies and sensory acceptance of Roselle have been reviewed to find out its current status. In advancing fermentation techniques, the food processor is faced with the challenge of modernizing the procedures and still conserving the traditional sensory characteristics of the Roselle products (particularly seasoning agents and wine) vital to consumer acceptance. Several indigenous traditional methodologies are difficult to apply on a large-scale by the food industries without changing the procedures which might end up with a product with unacceptable flavor. Thus, intensive studies and development activities are needed for these important products.

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109.

Index Note: Page numbers followed by “f” and “t” refer to figures and tables, respectively.

A Adriamycin, 165 Adverse effects, 194 African indigenous vegetables (AIVs), 1
2 Age, influence of, 52
53 AIVs. See African indigenous vegetables (AIVs) Amino acids, 79
85, 139 essential, 78
80, 82
83 Anemia treatment, 170
171 Anthocyanins, 48, 55
56, 144 anti-inflammatory activity, 160 activities, determination of, 207
209 as component of Hibiscus sabdariffa quality, 50
51 phytochemical compounds of, 93t Antidiabetic activity, 162
164 Antihyperlipidemic, 167
168 Antihypertensive activity, 159 Anti-inflammatory activity, 159
161 Antimicrobial activities, 51
52, 147 Antimicrobial potentials, 218
219 Antinutritional components, 54
55 Antiobesity activity, 161
162 Antioxidant activities, 202t, 204
205, 207
209, 208t Antioxidant properties, 146, 216
218 Ascorbic acid, 96
97 Atherosclerosis, 168
169

B Baked products, uses of Roselle in, 244
245 Beverages, 50, 246
247 Bikalga production, 235f Bioactive compounds, 57
59 therapeutic benefits of, 157
158 Bioactive constituents, 92 Breeding, 3
4

C Cadmium poisoning, 172
174 Calyces, 48, 70f, 80 antimicrobial activity, 147f cardioprotective activity of, 167 chemical composition of, 70
71, 71f fermented products, 240
243 harvesting of, 19
21, 20f, 21f

marketing of, 29 postharvest management of, 23, 24f, 25f proteins, processing of, 81 quality of proteins, 84 storage of, 27 volatile compounds of, 101t whole plant extracts, 124
129, 125f Cancer preventive activity, 171
172 Carbohydrates, 138
139 Cardioprotective activity, 167
169 antihyperlipidemic, 167
168 atherosclerosis, 168
169 Chemometrics analysis, 206
207, 206f Climate change, 5
8 Coleoptera, 36t

D Dietary fiber, 141
142 Diseases, 40
43 effects on Roselle, 41
42 prevention and control of, 42
43 chemical control, 42 nonchemical control, 42
43 types of, 40
41

E Ethnobotanical uses, 233t, 234

F Fatty acids, 116
120 Feeding values, 145
148 health benefits, 145
147 uses and value, 147
148 Fermented milk indigenous preferences, 248 uses of Roselle in, 243
244 Fermented studies, 234
245 baked products, 244
245 calyx’s fermented products, 240
243 fermented milk, 243
244 seeds fermented products, 234
240, 235f, 236f, 239f, 240f Fiber, 48 harvesting stems for, 22, 22f postharvest management of stems for, 26 whole plant extracts, 130
131

255

256

Index

Flavonoids, 143
144 activities, determination of, 207
209 antiobesity activity of, 161 phytochemical compounds of, 93t Food source, Roselle components as, 114
115

G General flowers, 80 Genetics diversity, 4
5

H Harvesting, 17
22, 52
53 of calyces, 19
21, 20f, 21f of leaves, 18
19, 19f stems for fiber, 22, 22f of tender shoots, 18
19, 19f Harvest maturity, 17 determinants of, 17
18 horticultural maturity, 17 physiological maturity, 17 timing of, 18t Health benefits, 145
147 antimicrobial activity, 147 antioxidant properties, 146 Heavy metals contamination, 8
9, 8f Hemiptera, 36t Hepatoprotective activity, 165
167 Hibiscus acid, 97
98, 98f Hibiscus-drug interactions, 189
190 Horticultural maturity, 17 Hydroxycitric acid, 97
98, 97f Hyperuricemia, 169
170

M Marketing, 28
29 of calyces, 29 of leaves, 29 of seeds, 29 of tender shoots, 29 Mbuja production, 238, 239f Methanolic extracts antidiabetic activity of, 162
163 anti-inflammatory properties of, 161 Microbial activities, 51
52 Microbial contamination, 8
9, 8f Minerals, 140, 140t, 141t Mycotoxins biocontrol agent against, 222
223 effect on human health, 220
222 production, and economic losses to food, 220, 221t

N Near-infrared (NIR) spectroscopy, 205, 205f Nephroprotective activity, 164
165 NIR. See Near-infrared (NIR) spectroscopy Nutraceutical, 78 Nutritional composition, 138
142 amino acids, 139 carbohydrates, 138
139 dietary fiber, 141
142 lipids, 139 minerals, 140, 140t, 141t proteins, 139 vitamins, 140
141, 142t

I Ikpiru processing, 240f

K Kidney functions, 193
194

L Leaves, 79
80 harvesting of, 18
19, 19f indigenous preferences, 248 marketing of, 29 postharvest management of, 23 proteins, processing of, 81
82 quality of proteins, 84 storage of, 27
28 whole plant extracts, 129 Lipase activity inhibition, 162 Lipids, 139 Liver functions, 193
194

O Oil recovery from seeds, 115
120 Oil seed products, 120 Organic acids, 92
98, 142
144, 143f, 143t anthocyanins, 144 ascorbic acid, 96
97 flavonoids, 143
144 hydroxycitric acid, 97
98, 97f Orthoptera, 36t Oxidative stability, 117
118, 120 Oxidative stress, 164

P Pests, 34
40 effects on Roselle, 35
38, 37f prevention and control of, 38
40 chemical control, 38
39 nonchemical control, 39
40

Index

types of, 34
35, 36t Pharmacological properties, 73
74 Phenolic acids, phytochemical compounds of, 93t Phenolic compounds, 119 Phenols activities, determination of, 207
209 physiological maturity (PM), 17, 52
53 Phytochemical compounds, 93t Pigments, composition of, 71
72 Plant growth, influence of, 52
53 PM. See physiological maturity (PM) Polyphenols, 157
158 anti-inflammatory activity, 160
161 cardioprotective activity of, 167 Postharvest management, 23
26 of calyces, 23
25, 24f, 25f of leaves, 23 of seeds, 25
26 of stems (fiber), 26 of tender shoots, 23 Protein products, 79
80 calyces, 80 general flowers, 80 leaves, 79
80 processing of, 81
82 calyces, 81 leaves, 81
82 quality of proteins, 82
84 seeds, 82 seeds, 79 Proteins, 139 concentrates, 78
79 fractions, 78
79 isolates, 78
79 of Roselle seeds, 115

Q Quality maintenance age, 52
53 anthocyanins, 50
51 antinutritional components, 54
55 harvesting, 52
53 microbial and antimicrobial activities, 51
52 plant growth, 52
53 prevailing factors, 54
55 secondary metabolites, 55
57 Quality of proteins, 82
84 calyces, 84 leaves, 84 seeds, 83
84

R RCR. See Roselle calyces residue (RCR) Renal effect, 169
170 Roselle (Hibiscus sabdariffa (H. sabdariffa) breeding, 3
4 in climate change, safe production of, 5
8 diseases of, 40
43 drink, 73f feeding values, 145
148 genetics diversity, 4
5 heavy metals contamination, 8
9, 8f interactions and toxicity, 189
194 measurement and maintenance of, 49
59 medicinal and therapeutic potential, 157
174 microbial contamination, 8
9, 8f nutritional properties, 138
144 pests of, 34
40 plant description, 69
70 products, 148
149 volatile compounds and phytochemicals of, 92
106 whole plant extracts, 123 Roselle calyces residue (RCR), 130
131

S Secondary metabolites, 55
57 Seeds, 79 fermented products, 234
240, 235f, 236f, 239f, 240f indigenous preferences, 247 marketing of, 29 oil recovery from, 115
120 postharvest management of, 25
26 protein of, 115 proteins, processing of, 82 quality of proteins, 83
84 storage of, 26f, 27
28 whole plant extracts, 129
130 Spectrophotometric techniques, 201
205 antioxidant activities, 202t, 204
205 total anthocyanin content, 204 total flavonoid content, 204 total phenolic content, 201
204 Stability, 116, 118
119 oxidation, 117
118, 120 Sterols, 116
117, 119 Storage, 27
28 of calyces, 27 of leaves, 27 of seeds, 26f, 27 of tender shoots, 27

257

258

Index

Streptozotocin (STZ) injection, 162, 164 STZ. See Streptozotocin (STZ) injection Synthetic fungicides, 222

U

T

V

TBT. See Triple bagging technology (TBT) Tender shoots harvesting of, 18
19, 19f marketing of, 29 postharvest management of, 23 storage of, 27
28 Therapeutic properties, 73
74 Tocopherols, 115
120 Total anthocyanin content, 204 Total flavonoid content, 204 Total phenolic content (TPC), 201
204 Toxicology, 191
192 TPC. See Total phenolic content (TPC) Traditional medicine, 158
159 Triple bagging technology (TBT), 43

Uricosuric effect, 165, 169
170 Utilization of products, 84
85

Vitamins, 140
141, 142t composition of, 72 Volatile compounds, 99
106, 101t

W Whole plant extracts, 123 calyx, 124
129, 125f leaves, 129 seeds, 129
130

Y Yanyanku processing, 240f Yogurt whole plant extracts, 131