(+)-Catechin and Quercetin from Pawpaw Pulp A
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Characterization of (+)-Catechin and Quercetin from Pawpaw Pulp A thesis presented to the faculty of the College of Health Sciences and Professions of Ohio University In partial fulfillment of the requirements for the degree Master of Science Jinsoo Ahn June 2011 © 2011 Jinsoo Ahn. All Rights Reserved. 2 This thesis titled Characterization of (+)-Catechin and Quercetin from Pawpaw Pulp by JINSOO AHN has been approved for the School of Applied Health Sciences and Wellness and the College of Health Sciences and Professions by Robert G. Brannan Assistant Professor of Applied Health Sciences and Wellness Randy Leite Interim Dean, College of Health Sciences and Professions 3 ABSTRACT AHN, JINSOO, M.S., June 2011, Human and Consumer Sciences, Food and Nutrition Characterization of (+)-Catechin and Quercetin from Pawpaw Pulp Director of Thesis: Robert G. Brannan This thesis investigates the concentration of total phenolics and total flavonoids in pulp extracts of pawpaw harvested in 2008, 2009, and 2010, and the concentration of (+)- catechin and quercetin flavonoids in 2010 pawpaw pulp extracts using high performance liquid chromatography (HPLC). Next, influence of frozen storage and air or vacuum packaging of pawpaw pulp on the concentration of (+)-catechin and quercetin flavonoids was examined. In addition, properties of pawpaw pulp such as moisture content, lipid content, percent sugar, color, and pH were measured. Total phenolics were determined using the Folin-Ciocalteu assay and reported as µmol gallic acid equivalent (GAE)/ g wet tissue. The concentration was observed in the order of 2009 sample (3.91 ± 1.61) < 2008 sample (11.19 ± 0.57) < 2010 sample (14.11 ± 1.90). Total flavonoids were determined spectrophotometrically using flavonoid rutin as a standard and reported as µmol rutin/ g wet tissue. The order of concentration was 2009 (0.46 ± 0.24) = 2010 (0.76 ± 0.40) < 2008 (2.01 ± 0.44). By using HPLC, the concentration of (+)-catechin and quercetin was determined as 0.43 ± 0.79 mg/ 100 g wet pulp and 0.46 ± 0.79 mg/ 100 g wet pulp, respectively. In both air and vacuum packaging under frozen condition, the concentration of (+)-catechin and quercetin increased during the first 4 months and decreased until the twelve-month period. In the meantime, an unknown compound was detected after quercetin was eluted. Pawpaw pulp has 75.24 % moisture, 0.45 % lipid, and 24.1 % 4 percent sugar and color was close to white, magenta, and yellow. The pH of pawpaw pulp was 4.85 which indicates slightly acidic. In conclusion, pawpaw pulp has considerable concentration of phenolic compounds which include flavonoids, and (+)-catechin and quercetin were major phenolic flavonoids in the pulp which increase their concentration during frozen storage before extraction. These results suggest that the two important flavonoids from pawpaw pulp can be essential sources for value-added functional food ingredients by optimization of storage condition of the pulp. Approved: _____________________________________________________________ Robert G. Brannan Assistant Professor of Applied Health Sciences and Wellness 5 To my late father, Professor Jong-Cheol Ahn 6 ACKNOWLEDGMENTS Foremost, I would like to express my sincere gratitude to my advisor Professor Robert G. Brannan, committee chairman for his continuous support for my master’s study and research. His guidance helped me during the entire time required to accomplish the research and write this thesis. Without his advice, this study would not have been successful. I also would like to acknowledge the members of my thesis committee, Professor Sharon Rana and Professor Michael Held, for their insightful comments and considerations. My special thanks go to Professor David H. Holben for all his encouragement, advice, and offering me a graduate assistantship. I also appreciate to Dr. Jennifer Horner, the Associate Dean of Research and Graduate Studies in the College of Health Sciences and Professions for proofreading and editing this thesis. In addition, I thank my fellow lab colleagues, other graduate students, and all the staff members in the School of Applied Health Sciences and Wellness and in the Food and Nutrition program. Last but not the least, I would like to thank my late father, mother, and brother for encouraging me to pursue this degree and supporting me throughout my life. 7 TABLE OF CONTENTS Page Abstract ............................................................................................................................... 3 Dedication ........................................................................................................................... 5 Acknowledgments .............................................................................................................. 6 List of Tables .................................................................................................................... 10 List of Figures ................................................................................................................... 11 CHAPTER 1: INTRODUCTION ..................................................................................... 12 Overview and Background ............................................................................................ 12 Statement of the Problem ............................................................................................... 14 Research Questions ........................................................................................................ 15 Significance of Research Study ..................................................................................... 15 Limitations ..................................................................................................................... 17 Glossary ......................................................................................................................... 17 CHAPTER 2: REVIEW OF CRITICAL LITERATURE ............................................... 19 Pawpaw .......................................................................................................................... 19 Background .............................................................................................................. 19 The Description and Horticulture of Pawpaw ......................................................... 20 Harvesting ................................................................................................................ 24 Postharvest Challenges ............................................................................................ 26 Characteristics of Pawpaw Fruit .............................................................................. 27 Sensory Characteristics ......................................................................................... 27 Nutritional Facts.................................................................................................... 28 Antioxidants .......................................................................................................... 29 Antioxidants in Fruits .................................................................................................... 30 Introduction .............................................................................................................. 30 Oxidation.................................................................................................................. 33 Oxidation in Biological Systems .......................................................................... 33 Oxidation in Foods ................................................................................................ 34 Types of Antioxidants .............................................................................................. 36 8 Flavonoids ...................................................................................................................... 38 Introduction .............................................................................................................. 38 Function ................................................................................................................... 39 Subgroups ................................................................................................................ 40 Daily Intake .............................................................................................................. 41 Flavonoid Subclasses ............................................................................................... 43 Flavonols ............................................................................................................... 43 Flavones ................................................................................................................ 44 Flavanones ............................................................................................................ 45 Flavan-3-ols .......................................................................................................... 46 Anthocyanidins ..................................................................................................... 47 Isoflavones ............................................................................................................ 48 Conclusion ..................................................................................................................... 49 CHAPTER 3: METHODOLOGY ...................................................................................