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Determination of Total Selenium and Seleno-Amino Acids in Yeast and Aquatic Organisms by Liquid Chromatography and Inductively Coupled Plasma Mass Spectrometry

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Determination of Total Selenium and Seleno-Amino Acids in Yeast and Aquatic Organisms by Liquid Chromatography and Inductively Coupled Plasma Mass Spectrometry DETERMINATION OF TOTAL SELENIUM AND SELENO-AMINO ACIDS IN YEAST AND AQUATIC ORGANISMS BY LIQUID CHROMATOGRAPHY AND INDUCTIVELY COUPLED PLASMA MASS SPECTROMETRY A Dissertation Presented to the Faculty of the Graduate School University of Missouri-Columbia In Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy by LILI WAN Dr. C. Michael Greenlief, Dissertation Supervisor DECEMBER 2007 The undersigned, appointed by the dean of the Graduate School, have examined the dissertation entitled DETERMINATION OF TOTAL SELENIUM AND SELENO-AMINO ACIDS IN YEAST AND AQUATIC ORGANISMS BY LIQUID CHROMATOGRAPHY AND INDUCTIVELY COUPLED PLASMA MASS SPECTROMETRY presented by Lili Wan a candidate for the degree of doctor of philosophy and hereby certify that, in their opinion, it is worthy of acceptance Professor C. Michael Greenlief Dr. William G. Brumbaugh Professor Silvia S. Jurisson Professor Susan Z. Lever Professor Sheryl A. Tucker Professor John C. Walker ii ACKNOWLEDGMENTS I would first like to personally express my sincere thanks to my mentor, Dr. C. Michael Greenlief, for the research opportunity, great advice, guidance and kind help throughout these years. I would like to thank Dr. William (Bill) G. Brumbaugh from Columbia Environmental Research Center (CERC). I really appreciate your encouragement and insightful comments for this research project. I would like to thank the staff in the branch of environmental chemistry at CERC, especially the assistance of Mike Walther, Jim Zajicek and Diane Nicks. I would like to thank my dear parents for their moral support and sweet love all these years. A special thanks goes to my dear brother for his kind comfort and taking care of our parents while I was not around. I would like to thank all my friends I have made here for their support, comfort and encouragement. I would like to thank Dr. Nathan D. Leigh and Dr. Zoltan Mester for their advice and help. I would like to thank the University of Missouri-Columbia for granting me the opportunity to study chemistry in the graduate department and for their funding though teaching assistantships. The funding for the work in this dissertation was provided in part by the U.S. Geological Survey, the U.S. Fish & Wildlife Service, and the Imperial Irrigation District, California. ii TABLE OF CONTENTS ACKNOWLEDGMENTS…………………………………………………………….......ii LIST OF TABLES…………………………………………………………………….….ix LIST OF FIGURES………………………………………………………………………xi ABSTRACT…………………………………………………………………………….xiii CHAPTER 1: SELENIUM 1.1 INTRODUCTION TO SELENIUM……………………………………………..…1 1.1.1 BACKGROUND OF RESEARCH………………………………………………1 1.2 CHEMISTRY AND BIOCHEMISTRY OF SELENIUM………..…..……………4 1.2.1 CHEMISTRY OF SELENIUM………………………………………...…...4 1.2.2 BIOCHEMISTRY OF SELENIUM………………………………………...5 1.3 SELENIUM IN THE ENVIRONMENT…………………………………………...5 1.3.1 SELENIUM IN WATERS…………………………………………………..6 1.3.2 SELENIUM IN SOILS……………………………………………………...7 1.3.3 SELENIUM IN PLANTS…………………………………………………...9 1.3.4 SELENIUM IN ANIMALS………………………………………………..11 1.4 SELENIUM IN HUMAN HEALTH……………………………………………...13 1.4.1 SELENIUM RETENTION AND METABOLISM………………………..13 1.4.2 SELENIUM TOXICITY AND DEFICIENCY……………………………15 1.5 SELENIUM IN FOOD ………………………………………………………..….16 1.5.1 INTRODUCTION..………………………………………………………..16 iii 1.5.2 SELENIUM IN BREAD AND CEREALS………………………………..17 1.5.3 SELENIUM IN BRAZIL NUTS…………………………………………..18 1.5.4 MEAT, EGGS, MILK AND FISH………………………………………...19 1.5.5 SELENIUM IN SELENIUM SUPPLEMENTS…………………………...20 1.6 REFERENCES……………………………………………………………………23 CHAPTER 2: ANALYTICAL TECHNIQUES FOR SELENIUM 2.1 INTRODUCTION TO ANALYTICAL TECHNIQUES FOR SELENIUM………………………………………………………………....31 2.2 SEPARATION TECHNIQUES…………………………………………………...31 2.2.1 INTRODUCTION…………………………………………………………31 2.2.2 HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)….…31 2.2.2.1 REVERSED-PHASE LIQUID CHROMATOGRAPHY………….….32 2.2.2.2 ION-PAIRING LIQUID CHROMATOGRAPHY……………………33 2.2.2.3 ION-EXCHANGE CHROMATOGRAPHY………………………….33 2.2.2.3.1 CATION-EXCHANGE CHROMATOGRAPHY………………..34 2.2.2.3.2 ANION-EXCHANGE CHROMATOGRAPHY…………………34 2.2.2.4 SIZE-EXCLUSION CHROMATOGRAPHY………………………...35 2.2.3 GAS CHROMATOGRAPHY……………………………………………..35 2.3 MASS SPECTROMETRY………………………………………………………..36 2.3.1 INTRODUCTION………………………………….……………………...36 2.3.2 IONIZATION SOURCES…………………………………………………38 2.3.2.1 INDUCTIVELY COUPLED PLASMA-MASS SPECTROMETRY (ICP-MS)………………..……………………….38 iv 2.3.2.2 ELECTROSPRAY IONIZATION-MASS SPECTROMETRY (ESI-MS)……………………………………………………………….39 2.3.2.3 MATRIX-ASSISTED LASER DESORPTION IONIZATION- MASS SPECTROMETRY (MALDI-MS)……………………………40 2.3.2.4 ELECTRON IMPACT (EI)………….………………………………..41 2.3.2.5 CHEMICAL IONIZATION (CI)…….………………………………..42 2.3.3 MASS SPECTROMETERS……………………………………………….42 2.3.4 TANDEM MASS SPECTROMETRY…………………………………….43 2.4 COUPLED TECHNIQUES……………………………………………………….44 2.4.1 INTRODUCTION…………………………………………………………44 2.4.2 LIQUID CHROMATOGRAPHY-INDUCTIVELY COUPLED PLASMA MASS SPECTROMETRY (LC-ICP-MS)…………..………....45 2.4.3 LIQUID CHROMATOGRAPHY-ELECTROSPRAY IONIZATION MASS SPECTROMETRY (LC-ESI-MS)……...…………46 2.4.4 GAS CHROMATOGRAPHY-MASS SPECTROMETRY (GC-MS)…….47 2.5 HYDRIDE GENERATION ATOMIC ABSORPTION SPECTROPHOTOMETRY (HG-AAS)…………………………………..………48 2.6 HYDRIDE GENERATION ATOMIC FLUORESCENCE SPECTROMETRY (HG-AFS)…………………………………………..……………………………..49 2.7 NEUTRON ACTIVATION ANALYSIS (NAA)………………………….……...50 2.8 REFERENCES………………...………………………………………………….52 CHAPTER 3: SELENIUM SPECIATION 3.1 INTRODUCTION TO SELENIUM SPECIATION…………………….………..56 3.2 SELENIUM SPECIATION IN YEASTS…………….…………………………..56 v 3.3 SELENIUM SPECIATION IN FOODS AND OTHER BIOLOGICAL SAMPLES………………………………….…………….………59 3.4 REFERENCES………….…………………………………………………….…..65 CHAPTER 4: INVESTIGATION OF DRC-ICP-MS FOR SELENIUM DETECTION 4.1 INTRODUCTION……………………………………………………...…………68 4.2 DETECTION OF TOTAL SELENIUM BY DRC- AND STANDARD ICP-MS…………………………………………………………….68 4.2.1 INTRODUCTION TO DRC-ICP-MS……………………………………..68 4.3 EXPERIMENTAL………………………………………………….……………..72 4.3.1 INSTRUMENTATION……………………………………………………72 4.3.2 OPTIMIZATION OF THE INSTRUMENTAL PARAMETERS………...73 4.3.3 REAGENTS AND MATERIALS…………………………………………74 4.3.4 QUALITY ASSURANCE…………………………………………………74 4.4 RESULTS AND DISCUSSION……………………………………….………….75 4.4.1 ELIMINATION OF POLYATOMIC INTERFERENCES………….…….75 4.4.2 ACCURACY OF ISOTOPE RATIO MEASUREMENTS IN ICP-MS…..77 4.4.3 CRITICAL EVALUATION OF DRC-ICP-MS…………………………...83 4.5 REFERENCES………………………………………………………………..…..85 CHAPTER 5: TOTAL SE DETERMINATION IN YEASTS AND AQUATIC ORGANISMS 5.1 INTRODUCTION………………………………………………………………...86 5.2 EXPERIMENTAL………………………………………….……………………..87 vi 5.2.1 INSTRUMENTATION AND REAGENTS……………………………….87 5.2.2 SAMPLE PREPARATION………………………………………………..88 5.2.3 SELENIUM MEASUREMENT WITH ON-LINE SIDA-ICP-MS……….89 5.3 INTRODUCTION OF STABLE ISOTOPE DILUTION ANALYSIS (SIDA)…..89 5.3.1 ANALYTICAL PROCEDURES FOR SELENIUM DETERMINATION WITH ON-LINE SIDA-ICP-MS…………………....90 5.3.2 ANALYSIS OF OLIGOCHAETES AND YEAST SAMPLES……….….96 5.3.3 ANALYSIS OF WHOLE DESERT PUPFISH…………………………..101 5.4 REFERENCES………………………………………………………….……….103 CHAPTER 6: DETERMINATION OF ORGANIC SELENO-AMINO ACIDS IN YEASTS AND AQUATIC ORGANISMS 6.1 INTRODUCTION……………………………………………………………….104 6.2 DETECTION OF SELENIUM COMPOUNDS BY LC-ICP-MS……………....104 6.2.1 INTRODUCTION……………………………………..…………………104 6.2.2 INSTRUMENTATION AND CHEMICALS……………….……………104 6.2.3 SAMPLE PREPARATION………………………………………………107 6.2.3.1 TRIS-BUFFER AQUEOUS EXTRACTION FOR WATER-SOLUBLE SELENO-AMINO ACIDS………...….……….107 6.2.3.2 ENZYMATIC DIGESTION FOR PROTEIN-BOUND SELENO-AMINO ACIDS………..…………………………………107 6.2.3.3 METHANESULFONIC ACIDIC DIGESTION FOR TOTAL SELENO-AMINO ACIDS…………………………………………..108 6.2.4 METHOD OPTIMIZATION……………………………………………..108 6.3 CONFIRMATION OF SELENIUM COMPOUNDS BY LC-ESI-MS AND MS/MS……………………………………...………………..111 vii 6.3.1 INTRODUCTION………………………………………………………..111 6.3.2 INSTRUMENTATION…………………………………………………..112 6.4 RESULTS AND DISCUSSION…………………………………...…………….113 6.4.1 CHROMATOGRAPHIC EVALUATION………………….……………113 6.4.2 CONFIRMATION OF SELENIUM COMPOUNDS BY LC-ESI-MS AND MS/MS………………………………………..….114 6.4.3 ANALYTICAL RESULTS FOR YEASTS ……………………………..117 6.4.4 ANALYTICAL RESULTS FOR WHOLE PUPFISH…………………...120 6.5 REFERENCES…………………………………………………………………..125 CHPTER 7: SUMMARY AND CONCLUSION…………………………………....126 Appendix A: SIDA-ICP-MS Selenium Calculation Worksheet……………………….128 Appendix B: SIDA-ICP-MS RESULTS OF TOTAL SELENIUM CONCENTRATION FOR LAB-CULTURED OLIGOCHAETES AND SELENIZED YEASTS………………..……..129 Appendix C: SIDA-ICP-MS RESULTS OF TOTAL SELENIUM CONCENTRATION FOR LAB-CULTURED PUPFISH……………....130 VITA……………………………………………………………………………………131 viii LIST OF TABLES Table Page Table 4.1: Spectral interferences of Se isotopes by ICP-MS……………………….….69 Table 4.2: Instrumental settings for ELAN DRC-e ICP-MS…………………………..73 Table 4.3: Apparent isotopic ratios based on net intensity ratios of 78Se/77Se and 80Se/77Se, and fraction of Se present as SeH+ based on the comparison of signals at mass 82 and 83………………………………………………..78 Table 5.1: SIDA-ICP-MS Selenium Calculation Worksheet………………………….95 Table 5.2: SIDA-ICP-MS results of total Se concentration for lab-cultured oligochaetes and selenized yeasts………………………………….....…....96 Table 5.3: Selenium analysis of biological certified reference materials with SIDA-ICP-MS…………………………………………………………......97 Table 5.4: Spike recoveries in selenized yeasts added before digestion…………...….98 Table 5.5: Se determinations of oligochaetes duplicates………………………………98 Table 5.6: Typical instrumental blanks and calibration verification results using SIDA-ICP-MS………………………………………………………..…....99
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