Chromatographic Methods in Clinical Chemistry and Toxicology

Editors ROGER L BERTHOLF, Ph.D., DABCC, Department of Pathology, University of Florida Health Science Center, Jacksonville, FL, USA RUTH E WINECKER, Ph.D., DABFT, North Carolina Office of the Chief Medical Examiner, Chapel Hill, NC, USA

• Preface xiList of Contributors xiii1. Quality Assurance, Quality Control and Method Validation in Chromatographic Applications 1Michele L. Merves and Bruce A. Goldberger1.1 Introduction 11.2 History 11.3 Definition of Quality Assurance and Quality Control 31.4 Professional Organizations 41.5 Internal Quality Assurance and Control 51.5.1 Standard operating procedure manual 51.5.2 Method development 51.5.3 Method validation 61.5.4 Accuracy 71.5.5 Precision 71.5.6 Recovery 71.5.7 Lower limits of detection (sensitivity) and quantitation 81.5.8 Range of linearity 81.5.9 Specificity 91.5.10 Stability 91.5.11 Carryover 91.5.12 Ruggedness 91.5.13 Selection of a reference standard 101.5.14 Selection of an internal standard and standard addition 101.5.15 Selection of derivatization agent 101.5.16 Selection of ions for selected-ion monitoring or full-scan analysis 111.5.17 Chromatographic performance 111.5.18 Statistical evaluation of quality control 111.6 External Quality Assurance 13References 132. Liquid Chromatographic-Mass Spectrometric Measurement of Anabolic Steroids 15Don H. Catlin, Yu-Chen Chang, Borislav Starcevic and Caroline K. Hatton2.1 Introduction 152.2 LC-MS Analysis of Synthetic Steroids or Animal Samples 162.3 LC-MS Analysis of Natural Androgens in Human Samples 192.4 Conclusion 29References 293. High-performance Liquid Chromatography in the Analysis of Active Ingredients in Herbal Nutritional Supplements 33Amitava Dasgupta3.1 Introduction 333.2 St John’s Wort 353.2.1 Drug interactions with St John’s wort 353.2.2 Measurement of active ingredients of St John’s wort using HPLC 363.2.3 Analysis of St John’s wort extract with other analytical techniques 383.2.4 Measurement of hypericin and hyperforin in human plasma using HPLC 383.3 Herbal Supplements with Digoxin-like Immunoreactivity 393.3.1 Use of HPLC for the determination of chan su, danshen and ginsengs 403.4 Herbal Remedies and Abnormal Liver Function Tests 413.4.1 Use of GC-MS and HPLC for the measurement of active components 433.5 Ginkgo Biloba 433.5.1 Analysis of components of ginkgo biloba by HPLC 443.6 Echinacea 453.6.1 Analysis of active components of echinacea by HPLC 453.7 Valerian 463.7.1 Analysis of components of valerian by HPLC 463.8 Feverfew 463.8.1 Analysis of parthenolide by HPLC 473.9 Garlic 473.9.1 Measurement of components of garlic by HPLC 483.10 Ephedra (Ma Huang) and Related Drugs 483.10.1 Analysis of active components of ephedra-containing products 493.11 Conclusions 50References 504. Measurement of Plasma L-DOPA and L-Tyrosine by High-Performance Liquid Chromatography as a Tumor Marker in Melanoma 56Thierry Le Bricon, Sabine Letellier, Konstantin Stoitchkov and Jean-Pierre Garnier4.1 Introduction 564.2 Melanogenesis 574.2.1 Overview of the pathway 574.2.2 Potential tumor markers 584.3 L-DOPA Alone 594.3.1 Urine analysis 594.3.2 Blood (plasma or serum) analysis 594.4 L-DOPA/L-Tyrosine Ratio 604.4.1 Technical aspects 604.4.2 Clinical results 614.4.3 Future directions 634.5 Conclusion 64References 655. Hypersensitive Measurement of Proteins by Capillary Isoelectric Focusing and Liquid Chromatography-Mass Spectrometry 67Feng Zhou and Murray Johnston5.1 Introduction 675.2 A Robust CIEF-RPLC Interface 695.3 First-Generation CIEF-RPLC-MS System for Proteins 715.4 Second-Generation CIEF-RPLC-MS System 765.5 Future Improvements 83Acknowledgment 83References 836. Chromatographic Measurement of Transferrin Glycoforms for Detecting Alcohol Abuse and Congenital Disorders of Glycosylation 87Anders Helander6.1 Introduction 876.2 Transferrin Microheterogeneity 886.3 Carbohydrate-deficient Transferrin (CDT) 896.4 Congenital Disorders of Glycosylation (CDG) 896.5 Analytical Methods for Transferrin Microheterogeneity 906.6 Chromatographic Methods for CDT 916.6.1 HPLC conditions and potential interferences 916.6.2 Chromatographic separation of transferrin glycoforms 926.6.3 Genetic transferrin variants and glycoform types 946.6.4 Sensitivity and reproducibility 946.7 Chromatographic Methods for CDG 946.7.1 HPLC testing for CDG 956.7.2 LC-MS testing for CDG 956.8 Summary and Conclusions 96References 977. Chromatographic Measurements of Catecholamines and Metanephrines 101Eric C. Y. Chan and Paul C. L. Ho7.1 Background 1017.1.1 Total or individual assays 1047.2 Analytical Measurements of Catecholamines and Metanephrines 1057.3 Early Methods 1057.3.1 Catecholamines 1057.3.2 Metanephrines 1067.4 Current Chromatographic Methods 1067.4.1 Chemistry of catecholamines 1067.4.2 Specimen preparation 1077.4.3 Fluorescence detection 1097.4.4 Electrochemical detection 1107.4.5 Chemiluminescence detection 1127.4.6 Mass spectrometry 1157.5 Practical Considerations for the Stability of Urinary Catecholamines and Metanephrines During Storage 1177.6 Future Developments 118Dedication 119References 1198. Chromatographic Measurement of Volatile Organic Compounds (VOCs) 127Larry A. Broussard8.1 Introduction 1278.2 General Considerations 1278.3 Intended Use 1288.4 Volatility of Compounds 1288.5 Sample Collection, Handling and Storage 1298.6 Headspace Gas Chromatographic Methods 1298.7 Columns and Detectors 1308.8 Identification, Quantitation and Confirmation 1308.9 Ethanol and Other Volatile Alcohols 1318.10 Inhalants and Screening for Multiple VOCs 1328.11 Interpretation 1348.12 Conclusion 136References 1369. Chromatographic Techniques for Measuring Organophosphorus Pesticides 139H. Wollersen and F. Musshoff9.1 Introduction 1399.2 Organophosphorus Pesticides (OPs) 1419.2.1 Mechanism of action 1419.2.2 Intoxication 1419.2.3 Progression of intoxication and longer term risks 1459.2.4 Therapy 1469.2.5 Analytical procedures 1469.3 Conclusion 163References 16410. Chromatographic Analysis of Nerve Agents 170Jeri D. Ropero-Miller10.1 Introduction 17010.2 Neuromuscular Blockers 17010.2.1 Background and uses 17010.2.2 Classification, mechanism and duration of action 17110.2.3 Effects and toxicity 17310.2.4 Analysis 17310.3 Paralytic Shellfish Poisoning: Saxitoxin 18510.3.1 Background 18510.3.2 Toxicity 18710.3.3 Analysis 18810.4 Summary 191References 19511. History and Pharmacology of c-Hydroxybutyric Acid 197Laureen Marinetti11.1 Introduction 19711.2 History of Illicit Use of GHB 19811.3 Clinical Use of GHB in Humans 20011.4 History of Illicit Use of GBL and 1,4BD 20011.5 Distribution and Pharmacokinetics of GHB, GBL and 1,4BD 20211.6 GHB Interpretation Issues and Post-mortem Production 20411.7 Analysis for GHB, GBL and 1,4BD 208References 21312. Liquid Chromatography with Inductively Coupled Plasma Mass Spectrometric Detection for Element Speciation: Clinical and Toxicological Applications 217Katarzyna Wrobel, Kazimierz Wrobel and Joseph A. Caruso12.1 Introduction 21712.2 Liquid Chromatography with Inductively Coupled Plasma Mass Spectrometric Detection 21812.3 Analytical Applications of Clinical and Toxicological Relevance 21912.3.1 Arsenic 21912.3.2 Iodine 23412.3.3 Mercury 23412.3.4 Platinum 24012.3.5 Selenium 24512.4 Conclusions and Future Trends 26012.5 Abbreviations 260References 26213. Applications of Gas Chromatography-Mass Spectrometry to the Determination of Toxic Metals 274Suresh K. Aggarwal, Robert L. Fitzgerald and David A. Herold13.1 Introduction 27413.2 Instrumentation 27513.3 Experimental Procedure 27613.3.1 Preparation of internal standard solutions 27613.3.2 Digestion of biological sample 27613.3.3 Preparation of metal chelate 27713.4 GC-MS Studies 27813.4.1 Memory effect evaluation 27813.4.2 Precision and accuracy in measuring isotope ratios 28113.4.3 Results of concentration determination of toxic metals in biological samples 28313.5 Conclusions 284References 284Index 287