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Antioxidant and Enzyme-Inhibitory Activities of Water-Soluble ANTIOXIDANT AND ENZYME-INHIBITORY ACTIVITIES OF WATER-SOLUBLE EXTRACTS FROM BANGLADESH VEGETABLES By Razia Sultana A thesis submitted to the Faculty of Graduate Studies, University of Manitoba In partial fulfillment of the requirements for the degree of Master of Science Department of Food and Human Nutritional Sciences University of Manitoba Winnipeg, Manitoba Canada Copyright © 2020 by Razia Sultana i ABSTRACT The interest in the possible health benefits of fruit and vegetable-rich diets is on the increase, partly because it has been recognized that increased consumption of vegetables may decrease the risk of developing non communicable diseases (NCDs) . The main attributes that can contribute to the antioxidants and enzyme inhibition properties of vegetables are polyphenolic compounds, minerals and vitamins that are present in abundance. Therefore, the aim of this thesis was to determine the NCD-related antioxidant and enzyme inhibition properties of nine water-soluble extracts of Bangladesh vegetables including ash gourd, bitter gourd, brinjal, Indian spinach, kangkong, okra, ridge gourd, snake gourd and stem amaranth. Aqueous polyphenol extracts were aquired to carry out all the assays because of the hydrophilic nature of the human gastrointestinal tract in addition to the high extraction yield and cheaper cost. The in vitro antioxidant and enzyme inhibition properties showed that almost all the vegtables were able to scavenge free radicals, chelate metal ions and reduce ferric ions, in addition to the inhibition of α-amylase, α-glucosidase, pancreatic lipase, angiotensin converting enzyme and renin. Indian Spinach, kangkong and okra showed the best activity to scavenge free radicals whereas Indian spinach and brinjal extracts performed better for inhibiting digestive enzymes responsible for the progression of NCDs. The quantities of amino acids, minerals and β-carotene available in these vegetables were also determined. ii ACKNOWLEDGEMENT First and foremost, I would like to thank the Almighty Allah for seeing me through the period of this program. I thank Allah for endowing me with strength, wisdom, knowledge and understanding to complete this program. I would like to express my deepest appreciation to my supervisor Dr. Rotimi Aluko for his continuous support and supervision and tremendous mentorship throughout this journey. I am thankful for the financial support from different sources such as University of Manitoba Entrance Graduate Student Scholarship, International Development Research Centre, Canada. I am also thankful to the Center for Natural Resources Studies(CNRS), Bangladesh and Dr. Emdad Haque for giving me this opportunity and serving on my program advisory committee. Additional thanks to Bangladesh Agricultural Research Institute (BARI), Bangladesh for providing me with the support to prepare my samples. I will like to thank Dr. Sijo Joseph who is also a member of my program advisory committee. An Enormous gratefulness to all the members of Dr. Aluko’s research group who contributed to the success of this Master’s degree specially Olayinka Ayo Oluwaguna for assisting me in different experiments. In a very special way, I would like to greatfully admire the continuous encouragement of Dr. Monisola Alashi, who trusts me always and provided mentorship in every way possible. She has taught me a lot on this journey. Finally, I want to thank my parents who sacrificed their life for me and supported me all my life. Thank you for your prayers and encouragement. I will always be grateful to both of you for standing iii beside me. My special thanks to my sisters who give me mental support always and encouraged me all the way. Lastly, my special gratitude to my brother-in-law who encouraged me to take this step. iv FOREWORD The manuscript format was followed in producing this Masters thesis.The work consists of two manuscripts bookended by a general introduction (Chapter1) and a literature review (Chapter 2) portion, and a conclusion part (Chapter 5). Each manuscript was prepared according to the style of the journal it is intended to be/has been published. Manuscript 1 (Journal of Food Biochemistry) deals with the Chemical composition and in vitro antioxidant properties of water-soluble extracts obtained from Bangladesh vegetables while manuscript 2 (“Foods” journal) examines the inhibitory activities of aqueous vegetable extracts against α-amylase, α-glucosidase, pancreatic lipase, renin, and angiotensin converting enzyme. A transition statement is added at the end of each manuscript and links it to the next chapter to provide coherence. Finally, the last chapter (Chapter 5) contains conclusions along with directions for future research. v TABLE OF CONTENTS ABSTRACT .................................................................................................................................................. ii ACKNOWLEDGEMENTS ......................................................................................................................... iii FOREWORD ................................................................................................................................................ v TABLE OF CONTENTS ............................................................................................................................. vi LIST OF TABLES ........................................................................................................................................ x LIST OF FIGURES ..................................................................................................................................... xi LIST OF ABBREVIATIONS ..................................................................................................................... xii CHAPTER ONE ........................................................................................................................................... 1 1.0 General Introduction ....................................................................................................................... 1 1.1 Hypotheses………………………………………………………………………………………………………………………………..6 1.2 Aim and Objectives ......................................................................................................................... 7 1.3 Role and contribution of the research to idrc-sponsored project……………………………………………….8 1.4 References ...................................................................................................................................... 9 CHAPTER TWO ........................................................................................................................................ 12 LITERATURE REVIEW ............................................................................................................. 12 2.1 Vegetables..................................................................................................................................... 12 2.1.1 Ash Gourd................................................................................................................................... 12 2.1.2 Bitter Gourd ............................................................................................................................... 13 2.1.3 Brinjal ......................................................................................................................................... 14 2.1.4 Indian Spinach ............................................................................................................................ 14 2.1.5 Kankgkong ................................................................................................................................. 14 2.1.6 Okra............................................................................................................................................ 15 2.1.7 Ridge Gourd ............................................................................................................................... 15 2.1.8 Snake Gourd ............................................................................................................................... 16 2.1.9 Stem Amaranth .......................................................................................................................... 16 2.2 Diet and health ............................................................................................................................. 17 2.3 Role of phytonutrients in preventing diseases associated with free radicals and oxidative stress ............................................................................................................................................................ 18 2.3.1 Free radical and oxidative stress ................................................................................................ 18 2.3.2 Sources of free radicals .............................................................................................................. 20 2.3.3 Deleterious effects of free radicals and oxidative stress ............................................................ 20 vi 2.4 Nutritional importance of vegetables ........................................................................................... 22 2.5 Importance of polyphenols as antioxidants .................................................................................. 25 2.5.1Phenolic compounds ................................................................................................................... 27 2.5.2 Carotenoids
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