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Comparative Stability of Phytonutrients In COMPARATIVE STABILITY OF PHYTONUTRIENTS IN FUNCTIONAL BEVERAGES STORED UNDER DIFFERENT ENVIRONMENTS THESIS Presented in Partial Fulfillment of the Requirements for the Degree Master of Science in the Graduate School of The Ohio State University By Shirley Yunita Witarsa, B.S. Graduate Program in Food Science and Technology The Ohio State University 2011 Thesis Committee: M. Monica Giusti, Advisor John Litchfield Michael Mangino Copyright by Shirley Yunita Witarsa 2011 Abstract The increasing consumer demand for foods that can enhance health, coupled to desire for lower sugar and calories intake has made functional beverages very popular. One major challenge in functional beverages development is that most nutrients and vitamins added are sensitive to the processing and storage conditions. A new system for delivery of phytonutrients into a beverage was evaluated which uses an enclosed chamber attached to the bottle (caps) to keep the ingredients in the dry form until released just prior to consumption. This experiment investigated the ability of these caps to deliver and preserve the chemical integrity of nutrients used in the beverage industry including ascorbic acid, hydroxycitric acid, riboflavin, pyridoxine, pyridoxal phosphate, niacinamide, and caffeine. Formulations containing known amounts of each nutrient were stored under four different conditions: in a cap attached to a bottle(1) or stored in a bag (2), hot-filled (3) and dissolved in water (4). Three different storage environments were applied: at 37˚C incubator without light exposure, at room temperature and exposed to light, at room temperature without light exposure. Samples were analyzed by HPLC-PDA-MS. Light exposure significantly reduced the stability of ascorbic acid, riboflavin, pyridoxine, and caffeine in commonly processed beverages. However, when ascorbic ii acid, riboflavin, pyridoxine, and caffeine stored in dry environment (inside a cap), significantly higher stability was obtained even under light exposure. Higher temperature storage significantly decreased the stability of ascorbic acid, hydroxycitric acid, and pyridoxal phosphate in commonly processed beverages. However, dry storage of those compounds resulted in higher stability under the same storage condition. No significant difference in niacinamide degradation rate among treatments in all storage conditions. There was also no significant difference of phytochemicals stability observed when stored in a cap off or on a bottled water. Light significantly affected the stability of betalains and anthocyanins when dissolved in water, resulting in clear or faded color solutions. Stability of these natural pigments increased when stored inside a cap. Our results indicate that the delivery of functional ingredients in the dry form inside a cap would improve stability, lower the manufacturing cost due to reduced losses, and provide functional beverages with more consistent nutritional value for customers. iii Acknowledgements The completion of this research reflects the supports and help of many people. I would like to thank Dr. M. Monica Giusti for the opportunity to join her lab, as well as for her direction, assistance, and guidance throughout the project. I would also like to acknowledge Liquid Health Labs, Inc. for their trust in allowing me to do this project and financial supports. I would like to thanks Dr. John Litchfield and Dr. Mike Mangino for their support and time in serving as my committee member. Additionally, I would like to thank my colleagues and friends: Scott Kottman, Kom Kamonpantana, Neda Ahmadiani, Steven Simmons, Allison Atnip, Abby Synder, Andrew Barry, Kumala Marthina, and Pauline Ie for their suggestions, help, and cooperation during our work in the lab. Lastly, I would like to thank my father Burhan Witarsa, mother Nieni Prajogo, and brother Darwin Witarsa, who trust me and support me in pursuing my Master’s Degree at The Ohio State University. iv VITA June 27, 1986 …………… Born in Semarang, Indonesia 2005-2009 ………………. B.S. in Food Science, Iowa State University, Ames, IA 2009-2011……………….. Graduate Research and Teaching Assistant The Ohio State University, Columbus, OH Field of Study Major Field: Food Science and Technology v Table of Contents Abstract ............................................................................................................................... ii Acknowledgements ............................................................................................................ iv VITA ................................................................................................................................... v List of Tables ..................................................................................................................... ix List of Figures ..................................................................................................................... x CHAPTER 1: INTRODUCTION ....................................................................................... 1 CHAPTER 2: LITERATURE REVIEW ............................................................................ 3 2.1 Functional Food .................................................................................................................. 3 2.2 Functional Beverages .......................................................................................................... 4 2.2.1 Energy drinks ................................................................................................................ 5 2.2.2 Enhanced water ............................................................................................................. 6 2.3 U.S functional beverages market ....................................................................................... 7 2.3.1 Market of functional beverages ..................................................................................... 7 2.3.2 Market for enhanced water and energy drink ................................................................ 8 2.4 Functionality of enhanced water and energy drink phytochemicals............................... 9 2.4.1 Vitamins ........................................................................................................................ 9 2.4.2 Energy Enhancing Ingredients .................................................................................... 10 2.4.3 Weight loss compounds ............................................................................................... 12 2.4.4 Natural food colorant .................................................................................................. 13 2.5 Stability of phytonutrients during Processing and Storage ........................................... 14 vi 2.5.1 Overview of phytonutrients stability ............................................................................ 14 2.5.2 Processing of functional beverage .............................................................................. 18 2.5.3 Packaging of Functional Beverage ............................................................................. 19 2.5.4 Nutrient Degradation .................................................................................................. 19 2.6 Optimizing the delivery of nutrients in functional beverages ........................................ 20 2.6.1 Vitamin Overage ......................................................................................................... 20 2.6.2 Active Packaging ......................................................................................................... 21 2.6.3 Dry storage ................................................................................................................. 22 CHAPTER 3: MATERIALS AND METHODOLOGY .................................................. 24 3.1 Materials............................................................................................................................ 24 3.2 Sample Preparation .......................................................................................................... 24 3.3 Accelerated Storage Study (30 days) ............................................................................... 25 3.4 Long term storage study ................................................................................................... 27 3.5 HPLC-MS analysis ........................................................................................................... 29 3.6 Color measurement .......................................................................................................... 32 3.7 Statistical analysis ............................................................................................................. 33 CHAPTER 4: RESULTS AND DISCUSSION................................................................ 34 4.1 Compounds separation and identification ...................................................................... 34 4.2 Composition of different enhanced water formulations ................................................ 37 4.3 Ascorbic acid stability ...................................................................................................... 42 4.4 Hydroxycitric acid stability .............................................................................................. 46 4.5 Riboflavin stability............................................................................................................ 48 vii 4.6 Pyridoxine and pyridoxal phosphate
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