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Adaptation and Validation of Food Product Specific Analytical Methods for Monitoring Prebiotics Present in Different Types of Processed Food Matrices

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Adaptation and Validation of Food Product Specific Analytical Methods for Monitoring Prebiotics Present in Different Types of Processed Food Matrices University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln Dissertations, Theses, & Student Research in Food Science and Technology Food Science and Technology Department 12-2011 Adaptation and Validation of Food Product Specific Analytical Methods for Monitoring Prebiotics Present in Different Types of Processed Food Matrices Rebbeca M. Duar University of Nebraska-Lincoln, [email protected] Follow this and additional works at: https://digitalcommons.unl.edu/foodscidiss Part of the Food Science Commons Duar, Rebbeca M., "Adaptation and Validation of Food Product Specific Analytical Methods for Monitoring Prebiotics Present in Different Types of Processed Food Matrices" (2011). Dissertations, Theses, & Student Research in Food Science and Technology. 21. https://digitalcommons.unl.edu/foodscidiss/21 This Article is brought to you for free and open access by the Food Science and Technology Department at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Dissertations, Theses, & Student Research in Food Science and Technology by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. ADAPTATION AND VALIDATION OF FOOD PRODUCT SPECIFIC ANALYTICAL METHODS FOR MONITORING PREBIOTICS PRESENT IN DIFFERENT TYPES OF PROCESSED FOOD MATRICES by Rebbeca M. Duar A THESIS Presented to the Faculty of The Graduate College at the University of Nebraska In Partial Fulfillment of Requirements For the Degree of Master of Science Major: Food Science and Technology Under the Supervision of Professor Vicki L. Schlegel Lincoln, Nebraska December, 2011 ADAPTATION AND VALIDATION OF FOOD PRODUCT SPECIFIC ANALYTICAL METHODS FOR MONITORING PREBIOTICS PRESENT IN DIFFERENT TYPES OF PROCESSED FOOD MATRICES Rebbeca M. Duar M.S University of Nebraska 2011 Adviser: Vicki L. Schlegel. Prebiotic carbohydrates are now added to a variety of processed foods to beneficially affect the gut microbial composition and activities. However, published data remain limited on the stability of prebiotics during food processes, such as baking, extrusion, pasteurization, high temperature heating, low pH condition, etc. As the complexity of the food matrix may also affect the ability to test for prebiotics, product specific analytical methods, including UV-vis spectroscopy, GC and HPLC, were developed and validated to monitor the stability of FOS, inulin and GOS in different types of processed food (breakfast cereal, cookie, muffin, sports drink and a nutritional bar). The results showed satisfactory linearity (r>0.9) for all the validated methods, however differences in the complexity of the ingredients were reflected in method accuracy and precision. Method precision was determined by calculating the percent relative standard deviation (% RSD) of spiked samples (n=5) below (0.5%) and above (2%) the target amount to be supplemented in the food products (1%) with results ranging from 1-39%. Methods for monitoring FOS and GOS resulted in low detection and quantitation limits allowing analysis of prebiotics of less than 1% in the presence of complex matrices. On the other hand the inulin method presented high detection and quantitation limits. Additionally, accuracy was affected by compounds present in the food matrix, which was accessed by determining the percent recoveries of spiked prebiotic in the control samples and applying corresponding correction factors to the supplemented processed foods when the accuracy was below 90% or above 110%. The chemical fate of the prebiotics was determined by applying the optimized and validated extraction and analytical method to prototype food products with 1% supplemented prebiotic. Recoveries ranged from 25-300% depending on method’s performance, complexity of the matrix and the severity of the processing effects. Finally the fate of the prebiotics in a cereal and sports drink that were prepared under various processing conditions indicated a high stability of GOS, while FOS and inulin were affected by low pH and high temperatures. iii ACKNOWLEDGMENTS First of all we thank USDA-NRI for funding this project. We also thank GTC nutrition, Beneo-Orafti, for providing us with prebiotics. This thesis wouldn’t be possible without the direction and dedication of my adviser, Dr Vicki Schlegel, to whom I extend my most sincere thanks for more than two years of mentoring, and support. My special thanks to Dr. Hutkins Dr. Wehling, Dr. Rose and Dr. Cuppett, for their help and advice. Also, thanks to Richard and all my lab mates for their help and good times in the lab. My deepest gratitude goes to my mom and aunt, without them I would have never made it this far in life. They have been there for me every step of the way, have always loved me unconditionally, and have supported me through all of my tough decisions. To my friends, my crowd, Felipe and Maria Isabel, thanks for your unconditional friendships and for company and support, especially late in the evenings, and when I needed help the most. Thanks guys for keeping me sane and happy through the journey of grad school. My special thanks to my “favorite” friend Michal, you never let me give up. Last but not least, thanks be to God for my life through all tests in the past two years. Nobody said it was easy, but at in the end you get what you deserve, the amount of effort you put determines the amount of joy that you receive. iv TABLE OF CONTENT LIST OF FIGURES ...................................................................................................................... vii LIST OF TABLES ......................................................................................................................... ix A. LITERATURE RIVIEW ............................................................................................................ 1 Background of Prebiotics ............................................................................................................. 1 A.1 Chemistry of Prebiotics ....................................................................................................... 1 A.1.1 Fructooligosaccharides ............................................................................................. 2 A.1.2 Inulin ......................................................................................................................... 4 A.1.3 Galactooligosaccharides ........................................................................................... 5 A.2 Health Promoting Properties of Prebiotics ......................................................................... 6 A.2.1 FOS Health Promoting Properties ............................................................................ 8 A.2.2 Inulin Health Promoting Properties ........................................................................ 10 A.2.3 GOS Health Promoting Properties ......................................................................... 12 A.3 Prebiotics in Foods ............................................................................................................ 15 A.3.1 FOS in Foods .......................................................................................................... 15 A.3.2 Inulin in Foods ........................................................................................................ 15 A.3.3 GOS in Foods .......................................................................................................... 19 A.4 Prebiotic Analysis ............................................................................................................. 21 A.4.1 FOS Analysis .......................................................................................................... 21 A.4.2 Inulin Analysis ....................................................................................................... 22 A.4.3 GOS Analysis ........................................................................................................ 22 B. OBJECTIVES AND SPECIFIC AIMS.................................................................................... 23 C. MATERIALS AND METHODS ............................................................................................. 24 C.1 Specific aim 1: Develop and/or adapt extraction and analytical procedures. ..................... 24 C.1.1. FOS Analysis ........................................................................................................... 27 C.1.2 Inulin Analysis .......................................................................................................... 29 C.1.3 GOS Analysis ........................................................................................................... 32 C.2 Specific Aim 2: Method Validation .................................................................................... 35 v C.2.1 Accuracy ................................................................................................................... 35 C.2.2 Precision ................................................................................................................... 35 C.2.3 Linearity of Calibration Curves ................................................................................ 35 C.2.4 Limit of Detection ..................................................................................................... 36 C.2.5 Limit of Quantitation ................................................................................................ 36 C.3 Specific Aim 3: Analysis of 1% prebiotic supplemented
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