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ABSTRACT MOHAMEDSHAH, ZULFIQAR YUSUF. Comparative ABSTRACT MOHAMEDSHAH, ZULFIQAR YUSUF. Comparative Assessment of Phenolic Bioaccessibility and Bioavailability from 100% Juice and Whole Fruit – A Preclinical Approach. (Under the direction of Dr. Mario Ferruzzi). The consumption of fruit is critical to meeting nutritional needs and in the prevention of chronic disease. As a nutrient dense food, fruit is a key source of micronutrients and dietary phytochemicals. However, less than a quarter of Americans consume the recommended daily fruit servings, driven by cost, convenience and other factors. 100% fruit juices are a cost effective and available option that accounts for ~35% of daily fruit consumption. Despite this, current dietary guidance favors consumption of whole fruit over 100% fruit juices for factors including loss of fiber and perceived increased sugar intake from juice products. However, there may be benefits to 100% fruit juice consumption as its’ liquid matrix, and the extensive mechanical and enzymatic action of the commercial juicing process, may provide an equivalent or better method for delivery of bioactive compounds, specifically phenolic species. By comparing phenolic-rich whole grapes and 100% grape juice, the objective of this research was to draw direct comparisons between fruit forms and investigate the impact of food matrix and processing (mastication of whole fruit vs. juice) has on bioavailability of phenolics and their secondary metabolites produced throughout human digestion. To understand if 100% grape juice can provide a matrix with highly bioaccessible phenolics relative to whole fruit, differences in phenolic content and bioaccessibility from commonly consumed table, Concord and Niagara grapes and their 100% juices were compared. Phenolic content in whole grapes and 100% juices were determined by LC-MS prior to in-vitro digestion to determine phenolic bioaccessibility. Concord and Niagara grape seeds had the highest concentration of phenolics in the forms of flavan-3-ols and larger procyanidins. Purple Concord grape skins were rich in anthocyanins and flavanols, while grape pulp had low quantities of phenolic species. While phenolic content of whole grapes was significantly (p < 0.01) greater than their 100% grape juices, following simulated digestion, absolute bioaccessible content of phenolics were found to be similar between grapes and 100% juice. Differences in bioaccessible content were driven by high relative bioaccessibility of anthocyanins in Concord juice compared to grapes as well as for flavan-3-ols and phenolic acids from grape juices to whole grapes. A greater fraction of skin and seed phenolics were extracted through juicing and made bioaccessible making 100% grape juice and whole fruit similar in overall phenolic delivery to consumers. In a second study, phenolic bioaccessibility and metabolism from Concord and Niagara grapes and corresponding 100% juices was assessed in both the upper and lower GI tract using an in-vitro digestion coupled with anaerobic gut fermentation model. Intestinal transport of resulting bioaccessible phenolics and metabolites was estimated using a Caco-2 cell model. Total bioaccessible phenolics from both upper and lower tract digestion was similar between whole grapes and 100% juices. Total cellular transport of phenolics was also similar between whole grapes and 100% juices. Some differences were observed between the location of phenolic metabolism, bioaccessibility and subsequent cellular transport of phenolics between grapes and juice. Specifically, greater amounts of flavonoids were transported from grape juices than whole grapes from the upper tract, which aligns with reported mechanisms for acute responses to grape juice consumption. Yet, cumulative bioaccessibility and transport from upper and lower GI digestion/fermentation together indicates that the absorbable phenolics from 100% grape juice is reflective of that of whole grapes, suggesting that phenolic-mediated health benefits from consumption of whole fruit and juice may be similar. These findings provide a mechanistic and compartmentalized framework that compares delivery characteristics of bioactive phenolics species between 100% grape juice and respective whole grapes. Furthermore, these results help to better understand the role 100% fruit juices play in a health promoting diet with respect to benefits from consuming fruits. © Copyright 2021 by Zulfiqar Yusuf Mohamedshah All Rights Reserved Comparative Assessment of Phenolic Bioaccessibility and Bioavailability from 100% Juice and Whole Fruit – A Preclinical Approach by Zulfiqar Yusuf Mohamedshah A thesis submitted to the Graduate Faculty of North Carolina State University in partial fulfillment of the requirements for the degree of Master of Science Food Science Raleigh, North Carolina 2021 APPROVED BY: _______________________________ _______________________________ Dr. Mario Ferruzzi Dr. Andrew Neilson Committee Chair _______________________________ Dr. Gabriel Harris ii DEDICATION To faith, family, and friends. iii BIOGRAPHY Zulfiqar Yusuf Mohamedshah was born on June 24th, 1996, in Montgomery, Maryland. He completed his Bachelor of Science in Chemistry at the University of Virginia in May 2018. Beginning in the summer of 2018, Zulfiqar worked as a Laboratory Technician under Dr. Mario Ferruzzi at the Plants for Human Health Institute in Kannapolis, North Carolina. Becoming intrigued by his work on the impacts of food processing on phenolic bioavailability, Zulfiqar transitioned to graduate studies under Dr. Ferruzzi. Upon completion of his master’s degree, Zulfiqar will pursue a doctorate in Chemistry at the University of California, San Diego. iv ACKNOWLEDGMENTS Firstly, I would like to thank Dr. Mario Ferruzzi for his constant and unyielding support, guidance, and encouragement. It has been an absolute privilege to be able to work with you. From you, I have not only learned how to conduct effective, meaningful research on the path to becoming a scientist, but also valuable life lessons and skills, and for that I am eternally grateful. I owe much of my successes and accomplishments to the opportunities you have afforded me and as I look to the next exciting chapter of my life and career, I can only hope to pay forward what you have done for me. I would like to thank my committee members, Dr. Andrew Nielson and Dr. Gabriel Harris for your insights, guidance, and support throughout my graduate work. To my lab mates, Micaela, Sydney, Candace, Hawi, Haley, Min, and Michael, I thank you for guidance, support, and encouragement throughout the years. Perhaps most importantly, thank you for putting up with my shenanigans. The long hours spent in lab would’ve been far less fun and educational without you all. To my friends and family, thank you for your constant support and interest in my work. My sanity and spirit for life and preserved because of you. I am not me without you. Lastly, and most importantly, I want to express my sincerest gratitude to my parents. It is difficult to express in mere words how grateful I am to have the two of you. Your love, grace, wisdom, and support are more than I deserve. I will never fully understand the sacrifices you have made for me, but nonetheless I am truly thankful. You have encouraged me to endlessly explore, question, and pursue whatever I fancy, making my education your priority. Thank you. v TABLE OF CONTENTS LIST OF TABLES ....................................................................................................................... vii LIST OF FIGURES .................................................................................................................... viii CHAPTER 1: Review of Relevant Literature ........................................................................... 1 1.1. Introduction ........................................................................................................................... 1 1.2. Introduction to Phenolics ................................................................................................... 12 1.3. Flavonoids ............................................................................................................................ 14 1.3.1. Anthocyanidins ...................................................................................................... 15 1.3.2. Flavones ................................................................................................................. 16 1.3.3. Isoflavones ............................................................................................................. 17 1.3.4. Flavan-3-ols ........................................................................................................... 17 1.3.5. Flavonols ................................................................................................................ 18 1.3.6. Flavanones ............................................................................................................. 19 1.4. Nonflavonoids ...................................................................................................................... 20 1.4.1. Hydroxycinnamates ............................................................................................... 21 1.4.2. Hydroxybenzoates.................................................................................................. 22 1.4.3. Stilbenes ................................................................................................................. 23 1.4.4. Lignans ..................................................................................................................
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