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Profiling Lipids for Authentication of High Value Ingredients by Mid-Infrared Spectroscopy Combined with Multivariate Analysis

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Profiling Lipids for Authentication of High Value Ingredients by Mid-Infrared Spectroscopy Combined with Multivariate Analysis Profiling Lipids for Authentication of High Value Ingredients by Mid-Infrared Spectroscopy Combined with Multivariate Analysis THESIS Presented in Partial Fulfillment of the Requirements for the Degree Master of Science in the Graduate School of The Ohio State University By Natalie E. Maurer Graduate Program in Food Science and Technology The Ohio State University 2012 Master's Examination Committee: Dr. Luis Rodriguez-Saona, Advisor Dr. Sheryl Barringer Dr. John Litchfield Copyrighted by Natalie E. Maurer 2012 Abstract Over the past decade, there has been an increase in demand for rapid techniques for authentication and detection of adulterants in food ingredients. Economic adulteration and counterfeiting of global food and consumer products is expected to cost the industry $10 to $15 billion per year. Verifying authenticity and detection of adulterants is something that can happen anywhere in the food supply chain, from the farmers to consumers and the industry to the manufacturer. The development of sensitive and robust screening tool(s) for assuring the quality of incoming raw materials would supplement the assurances provided by food manufacturer vendor auditing programs. The objective of this study was to develop a rapid and accurate method for the characterization and authentication of high valued ingredients. A temperature-controlled ZnSe ATR mid- infrared benchtop and diamond ATR mid-infrared portable handheld spectrometers were used to characterize sacha inchi oil and evaluate its oxidative stability compared to commercial oils. Soft independent model of class analogy (SIMCA) and partial least squares regression (PLSR) analyzed the spectral data. Fatty acid profiles showed that sacha inchi oil (44% linolenic acid) had similar levels of PUFA as flax oils. PLSR showed good correlation coefficients (R2>0.9) between reference tests and spectra from infrared devices, allowing for rapid determination of fatty acid composition and predicting oxidative stability. Oils formed distinct clusters allowing the evaluation of commercial sacha inchi oils from Peruvian markets and showed some prevalence of ii adulteration. Determining oil adulteration and quality parameters using the ATR-MIR portable handheld spectrometer allowed for portability and ease-of-use, making it a great alternative to traditional testing methods. Forty different cocoa butter samples encompassing an acceptable range of compositional variability for the chocolate industry were included. Cocoa butters were characterized for their melt characteristics (DSC Hardness), triacylglycerol content and fatty acid composition (GC-FAME). SIMCA and PLSR were used for classification and quantification analysis of cocoa butters. SIMCA classified all cocoa butters in distinct clusters in a 3-dimensional space but no sample clustering patterns were associated with melt characteristics. Spectral differences responsible for the separation of classes were attributed to stretching vibrations of the ester (–C=O) linkage (1660-1720 cm-1). PLSR models showed correlation coefficients > 0.93 and prediction errors (SECV) of 1.5 units for melt characteristics, 0.2-0.3% and 0.4- 0.8% for major fatty acids and triacylglycerols, respectively. ATR-IR spectroscopy combined with pattern recognition analysis provides robust models for characterization and determination of cocoa butter composition. Overall, FT-IR has proven to be a fast, reliable and highly reproducible method for the characterization and authentication of high valued ingredients. iii Dedicated to Joel. Thank you for always supporting me in following my dreams. iv Acknowledgments I would like to thank my advisor, Dr. Rodriquez-Saona for allowing me to work with him and for giving me the confidence I need in my abilities as a scientist. Thank you to Dr. Barringer and Dr. Litchfield for serving as committee members. Thank you to the members of my lab group, especially Emily Birkel and Ting Wang, who have always shown me great support and encouragement. v Vita June 2006 .......................................................Oak Hills High School May 2010 .......................................................B.S. Chemistry, Miami University 2010 to present ..............................................Graduate Research Associate, Department of Food Science and Technology, The Ohio State University Publications Characterization and Authentication of a Novel Vegetable Source of Omega-3 Fatty Acids, Sacha Inchi (Plukenetia volubilis L.) Oil Rapid Assessment of Quality Parameters in Cocoa Butter using FT-IR Spectroscopy and Multivariate Analysis Fields of Study Major Field: Food Science and Technology vi Table of Contents Abstract ............................................................................................................................... ii Dedication...........................................................................................................................iv Acknowledgments............................................................................................................... v Vita ..................................................................................................................................... vi List of Tables ................................................................................................................... ixx List of Figures ..................................................................................................................... x Chapter 1: Literature Review ............................................................................................. 1 1.1 Infrared Spectroscopy ............................................................................................... 1 1.2 Risk of Chemical Hazards ....................................................................................... 11 1.3 Vibrational Spectroscopy for Detection of Chemical Contaminants ...................... 17 1.4 References ............................................................................................................... 23 Chapter 2: Characterization and Authentication of a Novel Vegetable Source of Omega-3 Fatty Acids, Sacha Inchi (Plukenetia volubilis L.) Oil ..................................................... 28 2.1 Abstract ................................................................................................................... 29 2.2 Introduction ............................................................................................................. 30 vii 2.3 Materials and Methods ............................................................................................ 32 2.4 Results and Discussion ............................................................................................ 37 2.5 Conclusion ............................................................................................................... 53 2.6 Acknowledgement ................................................................................................... 53 2.7 References ............................................................................................................... 54 Chapter 3: Rapid Assessment of Quality Parameters in Cocoa Butter using ATR-IR Spectroscopy and Multivariate Analysis .......................................................................... 57 3.1 Abstract ................................................................................................................... 58 3.2 Introduction ............................................................................................................. 59 3.3 Materials and Methods ............................................................................................ 62 3.4 Results and Discussion ............................................................................................ 67 3.5 Conclusion ............................................................................................................... 76 3.6 References ............................................................................................................... 78 List of References ............................................................................................................. 80 viii List of Tables Table 2.1. Fatty acid composition for commercial vegetable oils (olive, canola, cottonseed, corn, high oleic sunflower, sunflower, flax and sacha inchi) and pure authentic sacha inchi oils using fatty acid methyl ester (FAME) procedure............................................................................................................................39 Table 2.2. PLSR model statistical analysis for determining fatty acid composition in oils (corn, high oleic sunflower, flax and sacha inchi) using a benchtop and hendheld infrered system ................................................................................................................................45 Table 2.3. Peroxide value (PV), free fatty acid (FFA), and fatty acid composition for the different oil samples studied during a 20 day oxidative stability test................................49 Table 2.4. Statistical analyses for the PLSR models developed to determine the PV and FFA of oils (corn, high oleic sunflower, flax and sacha inchi) during their oxidation process using a mid-IR ATR benchtop and mid-IR ATR portable handheld spectrometers.....................................................................................................................51 Table 3.1. Statistical data for
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