East Tennessee State University Digital Commons @ East Tennessee State University Electronic Theses and Dissertations Student Works 8-2009 Comparison of Artificial Flavors in Commercial Products and Actual Natural Flavor via Gas Chromatography Mass Spectroscopy Data. Randi Jasmine Sluss East Tennessee State University Follow this and additional works at: https://dc.etsu.edu/etd Part of the Food Chemistry Commons Recommended Citation Sluss, Randi Jasmine, "Comparison of Artificial Flavors in Commercial Products and Actual Natural Flavor via Gas Chromatography Mass Spectroscopy Data." (2009). Electronic Theses and Dissertations. Paper 1804. https://dc.etsu.edu/etd/1804 This Thesis - Open Access is brought to you for free and open access by the Student Works at Digital Commons @ East Tennessee State University. It has been accepted for inclusion in Electronic Theses and Dissertations by an authorized administrator of Digital Commons @ East Tennessee State University. For more information, please contact [email protected]. Comparison of Artificial Flavors in Commercial Products and Actual Natural Flavor Via Gas Chromatography Mass Spectrometry Data A thesis presented to the faculty of the Department of Chemistry East Tennessee State University In partial fulfillment of the requirements for the degree Master of Science in Chemistry by Randi Jasmine Sluss August 2009 Dr. Chu-Ngi Ho, Chair Dr. Peng Sun Dr. Ningfeng Zhao Keywords: Strawberry, Gas Chromatography Mass Spectroscopy, Natural Flavor, Artificial Flavor ABSTRACT Comparison of Artificial Flavors in Commercial Products and Actual Natural Flavor Via Gas Chromatography Mass Spectroscopy Data by Randi Jasmine Sluss In this research project, real natural strawberries bought from different local sources were profiled by gas chromatography-mass spectroscopy (GCMS). These profiles were then used as a reference to compare GCMS profiles of commercial artificial strawberry flavor products such as strawberry flavored Cool SplashersTM, Gatorade®, and Aquacal® flavored water. The chromatograms obtained were patterned using simple visual observations, scatter plot designs, Mann-Whitney U Test, and correlation coefficients. The artificially flavored commercial products tend to have simpler chromatograms. The Burger King® milkshake, Gatorade®, and Hi- C® are the most similar to that of the natural strawberry flavor. Their correlation coefficients are 0.972, 0.870, and 0.984 respectively. The Mann-Whitney U Test results also support the conclusions from correlation coefficients. However, the natural products tend to have more constituents including the main flavoring compounds. Thus fresh produce have better flavor and are more nutritious for a good reason. 2 DEDICATION I would like to dedicate this thesis to everyone who believes in me and gives me the strength to accomplish all my goals. I dedicate this thesis to a family who believes in me and gives me the courage to push through struggles in my life. I dedicate this thesis to the entire faculty of the Chemistry Department at ETSU who believes in me and pushes me to do better. I dedicate this thesis to God who also gives me the strength and courage to accomplish all my goals. 3 ACKNOWLEDGMENTS I would like to acknowledge the entire ETSU faculty that aided me in the process of completing all the requirements for graduation and my thesis. I would like to thank ETSU for awarding me the graduate assistantship scholarship. 4 CONTENTS Page ABSTRACT…………………………………………………………………………………… 2 DEDICATION………………………………………………………………………………… 3 ACKNOWLEDGEMENTS…………………………………………………………………… 4 LIST OF TABLES……………………………………………………………………………. 7 LIST OF FIGURES…………………………………………………………………………… 8 Chapter 1. INTRODUCTION……………………………………………………………………. 9 Origins of Foods……………………………………………………………….. 9 Food Additives…………………………………………………....................... 11 Natural and Artificial Flavors…………………………………………………. 13 Strawberries…………………………………………………………………… 18 2. METHODS OF FLAVOR ANALYSIS……………………………………………… 21 GC/MS Methods………………………………………………………………. 21 Flavor Compounds Extraction Methods……………………………………… 23 Solvent Liquid/Liquid Extraction…………………………………….. 23 Static Headspace Sampling…………………………………………… 24 Dynamic Headspace Sampling……………………………………….. 25 Solid Phase Micro-Extraction(SPME)……………………………….. 25 Soxhlet Extraction……………………………………………………. 25 Supercritical Fluid Extraction……………………………………….. 26 Simultaneous Steam Distillation/Extraction…………………………... 26 5 Molecular Distillation…………………………………………………. 27 Flavor Analysis……………………………………………………………….. 28 Proposed Research……………………………………………………………. 34 3. EXPERIMENTAL PROCEDURES…………………………………………………. 36 Reagents………………………………………………………………………. 36 Samples Obtained…………………………………………………………….. 36 Preparation of Standard Solutions…………………………………………….. 37 Preparation of Natural Strawberry Samples…………………………………… 38 Preparation of Commercial Products Samples……………………………….... 38 Instrumentation……………………………………………………………….. 40 Data Analysis…………………………………………………………………. 43 4. RESULTS AND DISCUSSION……………………………………………………… 45 Visual Inspection of GCMS Chromatograms………………………………… 45 Scatter Plot Study……………………………………………………………. 54 Statistical Methods.…………………………………………………………… 57 5. CONCLUSION………………………………………………………………………. 62 REFERENCES………………………………………………………………………. 67 VITA…………………………………………………………………………………. 71 6 LIST OF TABLES Table Page 1. Flavor Compounds and Contributed Flavors…….………………………………… 16 2. Important Compounds Found in Strawberries……………………………………… 20 3. HP 5890 GC Parameters……………………………………………………………. 4q 4. HP 5971 Mass Selective Detector Parameters……………………………………… 42 5. Correlation Coefficients of the Different Natural Strawberry Samples Obtained From GCMS Data…………………………………………………………………............. 57 6. Correlations Coefficients Between the Natural Strawberry Samples and the Commercial Samples……………………………………………………………………………… 58 7. Mann-Whitney U Test for Comparison of Natural Strawberry vs. Commercial Products………………………………………………………………. 61 7 LIST OF FIGURES Figure Page 1. GCMS a Simple Schematic of a Gas Chromatograph Mass Spectrometer..................... 22 2. GCMS Spectrum of Standards, Ethyl Acetate, Ethyl Butyrate, Furfural, and DMHF of Concentrations of 1.02 x 10-4 , 7.57 x 10-5, 1.21 x 10-4, and 7.80 x 10-6 mol/L, Repectively…………………………………………………………...……… 45 3. GCMS Spectrum of California Strawberry Sample of Concentration of 0.1 g/mL….. 46 4. GCMS Spectrum of Driscoll’s Strawberry Sample of Concentration of 0.1 g/mL….. 47 5. GCMS Spectrum of Strawberry Fraises Sample of Concentration of 0.1 g/mL……... 48 6. GCMS Spectrum of Burger King® Milkshake Sample of Concentration 0.4 g/mL…. 49 7. GCMS Spectrum of Gatorade® Sample of Concentration 0.03 g/mL..………………. 49 8. GCMS Spectrum of Hi-C® Sample of Concentration 0.11 g/mL…………………….. 51 9. GCMS Spectrum of JELL-O® Sample of Concentration 0.03 g/mL…………………. 52 10. GCMS Spectrum of Aquacal® Sample of Concentration 0.05 g/mL…………………. 52 11. GCMS Spectrum of Cool SplashersTM Sample of Concentration 0.02 g/mL………… 53 12. Scatter Plot of Real Strawberry Averaged Retention Times Compared to Those of the Commercial Products………………………………………………………………... 55 13. Scatter Plot of Real Strawberry Averaged Retention Times Compared to Those of the Commercial Products………………………………………………………………… 56 8 CHAPTER 1 INTRODUCTION Origins of Foods Foods we consume are regulated by the Food and Drug Administration (FDA). Foods can contain carbohydrates, fats, proteins, sugars, fiber, salts, vitamins, minerals, antioxidants, phytonutrients, and food additives. Many of these components are used to develop the commercial foods we eat today (1). The origins of foods are quite different. It is found that most of our predecessors ate wild foods consisting of seeds, nuts, fruits, roots, insects, and honey. By eating the same plants and fruits as the surrounding animals, humans could deduce which foods were poisonous and which were safe to eat. Animals were also hunted and consumed as a primary source of protein. Further, it has also been discovered that humans were able to hunt larger animals as far back as nearly two million years ago. Although, the use of fire to cook the meat was dated back to about 20,000 years ago, the first recorded use of fire was 30,000 years earlier. However, its purpose was not to cook food. Rather, fire was used for warmth and often to scare animals away. This time was known as the Ice Age. As this era ended, nearly 10,000 years later, humans used fire extensively as a light source to hunt animals. The animals that were targeted were often overhunted; leading to a shortage in the food supply. This situation initiated the origins of agriculture (2). Nearly 10,000 years ago, these agricultural beginnings brought about the early stages of food organization. The cultivating of seeds and other plants began as humans discovered the most fertile times of the year in which to plant their crops. Some of these species of plants are still grown today. Among the few crop sources of this time, wheat was cultured in Persia and 9 Afghanistan, along with other grains in Ethiopia. Migration brought wheat into Europe as oats, barley, and rye grew more effectively in the northern lands. Further, there are recorded instances of beer and liquor made from these grains as early as 5000 years ago. Yeast, recorded as an early food additive, made this transition possible with the development of bread soon to follow. Rice and maize were next in line to be grown extensively in both the Far East and Central America. Turnips, radishes, almonds, peaches, apricots, beetroots, cabbages, onions,