TM M E T H O D S I N MO L E C U L A R B I O L O G Y Series Editor John M. Walker School of Life Sciences University of Hertfordshire Hatfield, Hertfordshire, AL10 9AB, UK For other titles published in this series, go to www.springer.com/series/7651 Clinical Applications of Mass Spectrometry Methods and Protocols Edited by Uttam Garg Children’s Mercy Hospitals and Clinics, Kansas City, MO, USA Catherine A. Hammett-Stabler University of North Carolina, Chapel Hill, NC, USA Editors Uttam Garg Catherine A. Hammett-Stabler Department of Pathology & Laboratory Department of Pathology & Laboratory Medicine Medicine Children’s Mercy Hospitals and Clinics University of North Carolina 2401 Gillham Road Chapel Hill, NC 27599-7525 Kansas City, MO 64108 USA USA Dedicated to Jyotsna Garg and Thomas Stabler for their patience and understanding throughout this process. Preface No longer considered an esoteric technique, mass spectrometry has entered the clinical laboratory where it is being used for a wide range of applications. Methods developed using this technique offer a level of specificity and sensitivity unrealized by spectro- photometric- and immunoassay-based methods. In fact, the technique is considered essential coupled to either gas chromatography or liquid chromatography for the measurement of many clinical analytes. This volume, Clinical Applications of Mass Spectrometry, is intended to provide detailed step-by-step procedures for the analysis of number of analytes of clinical importance. Each chapter provides the brief introduction of the analyte followed by a detailed analytical protocol. The procedures are easy to follow and should be easily reproducible in the laboratory already using mass spectrometry and wanting to intro- duce new analyses as well as those laboratories considering adding the technology. We do not offer a detailed overview of mass spectrometry since there are many excellent volumes and tutorials on MS theory and principles, but we have included a chapter that discusses some of the basic laboratory practices to remember when introducing the technique into routine use. The analytes for which mass spectrometry methods are described include drugs, hormones, and metabolic compounds spanning the disciplines of toxicology, therapeutic drug monitoring, endocrinology, and pediatric metabolism. Demonstrating the versatility of the technique, we have been able to provide multiple applications for some analytes so that the reader has his/her choice of an LC-MS- or a GC-MS-based method. We thank all of our colleagues who contributed the methods. We realize that the development of these represent thousands of hours of work. And we hope that those who use this book find it useful. Uttam Garg Catherine A. Hammett-Stabler vii Contents Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii Contributors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xiii 1. The Evolution of Mass Spectrometry in the Clinical Laboratory. . . . . . . . . . . . . . . 1 Catherine A. Hammett-Stabler and Uttam Garg 2. Measurement of Plasma/Serum Acylcarnitines Using Tandem Mass Spectrometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 Nathalie Lepage and Susan Aucoin 3. Comprehensive Determination of Amino Acids for Diagnosis of Inborn Errors of Metabolism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27 Dennis J. Dietzen and Annette L. Weindel 4. Identification and Quantitation of Amphetamine, Methamphetamine, MDMA, Pseudoephedrine, and Ephedrine in Blood, Plasma, and Serum Using Gas Chromatography-Mass Spectrometry (GC/MS) . . . . . . . . . . . . . . . . . . . . . . . . . . 37 Josh Gunn, Scott Kriger, and Andrea R. Terrell 5. Quantification of Antidepressants Using Gas Chromatography-Mass Spectrometry. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45 Ruth E. Winecker 6. Quantitation of Argatroban in Plasma Using Liquid Chromatography Electrospray Tandem Mass Spectrometry (UPLC-ESI-MS/MS) . . . . . . . . . . . . . . 57 Ross J. Molinaro 7. Quantitation of Amobarbital, Butalbital, Pentobarbital, Phenobarbital, and Secobarbital in Urine, Serum, and Plasma Using Gas Chromatography-Mass Spectrometry (GC-MS) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65 Leonard L. Johnson and Uttam Garg 8. Quantitation of Benzodiazepines in Blood and Urine Using Gas Chromatography-Mass Spectrometry (GC-MS) . . . . . . . . . . . . . . . . . . . . . . . . . . . 75 Bruce A. Goldberger, Chris W. Chronister, and Michele L. Merves 9. LC-MS/MS Analysis of 13 Benzodiazepines and Metabolites in Urine, Serum, Plasma, and Meconium. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89 Stephanie J. Marin and Gwendolyn A. McMillin 10. Simultaneous Determination and Quantification of 12 Benzodiazepines in Serum or Whole Blood Using UPLC/MS/MS . . . . . . . . . . . . . . . . . . . . . . . . . 107 Josh Gunn, Scott Kriger, and Andrea R. Terrell 11. Determination of Benzoic Acid in Serum or Plasma by Gas Chromatography-Mass Spectrometry (GC/MS) . . . . . . . . . . . . . . . . . . . . . . . . . . 121 Jeff M. Knoblauch, David K. Scott, Laurie D. Smith, and Uttam Garg 12. Quantification of Busulfan in Plasma Using Liquid Chromatography Electrospray Tandem Mass Spectrometry (HPLC-ESI-MS/MS) . . . . . . . . . . . . . . 129 Marion L. Snyder and James C. Ritchie ix x Contents 13. Quantitation of Total 11-Nor-9-Carboxy-Delta 9-Tetrahydrocannabinol in Urine and Blood Using Gas Chromatography-Mass Spectrometry (GC-MS). . . 137 C. Clinton Frazee III, Michael Kiscoan, and Uttam Garg 14. Quantitation of Cocaine, Benzoylecgonine, Ecgonine Methyl Ester, and Cocaethylene in Urine and Blood Using Gas Chromatography-Mass Spectrometry (GC-MS) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 145 Steven W. Fleming, Amitava Dasgupta, and Uttam Garg 15. Identification and Quantitation of Cocaine, Benzoylecgonine, and Cocaethylene in Blood, Serum, and Plasma Using Ultra-Performance Liquid Chromatography Coupled to Tandem Mass Spectrometry (UPLC-MS/MS) . . . . . . . . . . . . . . . . . . 157 Scott Kriger, Josh Gunn, and Andrea R. Terrell 16. Detection and Quantification of Cocaine and Benzoylecgonine in Meconium Using Solid Phase Extraction and UPLC/MS/MS . . . . . . . . . . . . . . . . . . . . . . . . 165 Josh Gunn, Scott Kriger, and Andrea R. Terrell 17. Diagnosis of Creatine Metabolism Disorders by Determining Creatine and Guanidinoacetate in Plasma and Urine. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 175 Qin Sun and William E. O’Brien 18. Broad Spectrum Drug Screening Using Electron-Ionization Gas Chromatography-Mass Spectrometry (EI-GCMS) . . . . . . . . . . . . . . . . . . . . . . . . . 187 Judy Stone 19. Broad Spectrum Drug Screening Using Liquid Chromatography-Hybrid Triple Quadrupole Linear Ion Trap Mass Spectrometry . . . . . . . . . . . . . . . . . . . . . 203 Judy Stone 20. High Sensitivity Measurement of Estrone and Estradiol in Serum and Plasma Using LC-MS/MS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 219 Mark M. Kushnir, Alan L. Rockwood, Bingfang Yue, and A. Wayne Meikle 21. 3-Hydroxy-Fatty Acid Analysis by Gas Chromatography-Mass Spectrometry. . . . . 229 Patricia M. Jones and Michael J. Bennett 22. Quantitation of Fentanyl in Blood and Urine Using Gas Chromatography-Mass Spectrometry (GC-MS) . . . . . . . . . . . . . . . . . . . . . . . . . . . 245 Bruce A. Goldberger, Chris W. Chronister, and Michele L. Merves 23. Determination of Total Homocysteine in Plasma Using Liquid Chromatography Coupled to Tandem Mass Spectrometry (LC/MS/MS) . . . . . . 253 Bruno Casetta 24. Quantitation of Homovanillic Acid (HVA) and Vanillylmandelic Acid (VMA) in Urine Using Gas Chromatography-Mass Spectrometry (GC/MS). . . . . . . . . . . 261 Ryan Allenbrand and Uttam Garg 25. Quantitation of 17-OH-Progesterone (OHPG) for Diagnosis of Congenital Adrenal Hyperplasia (CAH) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 271 Ravinder J. Singh 26. Determination of Hydroxyurea in Serum or Plasma Using Gas Chromatography-Mass Spectrometry (GC-MS) . . . . . . . . . . . . . . . . . . . . . . . . . . . 279 David K. Scott, Kathleen Neville, and Uttam Garg 27. Quantitation of Ibuprofen in Blood Using Gas Chromatography-Mass Spectrometry (GC-MS) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 289 Gerry Huber and Uttam Garg Contents xi 28. Quantitation of Indomethacin in Serum and Plasma Using Gas Chromatography-Mass Spectrometry (GC-MS) . . . . . . . . . . . . . . . . . . . . . . . . . . . 297 Sarah Thomas, Amanda Sutton, and Uttam Garg 29. Quantitation of Iothalamate in Urine and Plasma Using Liquid Chromatography Electrospray Tandem Mass Spectrometry (HPLC-ESI-MS/MS) . . . . . . . . . . . . . . 307 Ross J. Molinaro and James C. Ritchie 30. Identification and Quantitation of Ketamine in Biological Matrices Using Gas Chromatography-Mass Spectrometry (GC-MS) . . . . . . . . . . . . . . . . . . 317 Timothy P. Rohrig, Megan Gamble, and Kim Cox 31. Measurement of Filter Paper Bloodspot Lead by Inductively Coupled Plasma-Mass Spectrometry (ICP-MS). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 327 Denise M. Timko and Douglas F. Stickle 32. Multiplex Lysosomal Enzyme Activity Assay on Dried Blood Spots Using Tandem Mass Spectrometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 339 X. Kate Zhang, Carole S. Elbin, Frantisek Turecek, Ronald Scott, Wei-Lien Chuang, Joan M. Keutzer, and Michael Gelb 33. Gas Chromatography-Mass Spectrometry Method for the Determination of Methadone and 2-Ethylidene-1,5-Dimethyl-3, 3-Diphenylpyrrolidine (EDDP) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 351 Christine L.H. Snozek, Matthew W. Bjergum, and Loralie J. Langman 34. Liquid Chromatography-Tandem Mass Spectrometry (LC-MS-MS) Method for Monitoring Methotrexate in the Setting of Carboxypeptidase-G2 Therapy. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 359 Vikram S. Kumar, Terence Law, and Mark Kellogg 35. Quantitation of Methylmalonic Acid in Serum or Plasma Using Isotope Dilution-Selected Ion Gas Chromatography-Mass Spectrometry . . . . . . . . . . . . . . 365 Jie Chen and Michael J. Bennett 36. Quantitation of Methylmalonic Acid in Plasma Using Liquid Chromatography – Tandem Mass Spectrometry . . . . . . . . . . . . . . . . . . . . . . . . . . . 371 Claudine Fasching and Jasbir Singh 37. Monitoring of Mycophenolate Acid in Serum or Plasma Using LC Tandem Mass Spectrometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 379 Catherine A. Hammett-Stabler, Diane Ciuffetti Geis, James C. Ritchie, and Christine Papadea 38. Quantitation of Nicotine, Its Metabolites, and Other Related Alkaloids in Urine, Serum, and Plasma Using LC-MS-MS. . . . . . . . . . . . . . . . . . . . . . . . . . . 389 Bingfang Yue, Mark M. Kushnir, Francis M. Urry, and Alan L. Rockwood 39. Quantitation of Opioids in Blood and Urine Using Gas Chromatography-Mass Spectrometry (GC-MS) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 399 Bruce A. Goldberger, Chris W. Chronister, and Michele L. Merves 40. Quantitation of Morphine, Codeine, Hydrocodone, Hydromorphone, Oxycodone, Oxymorphone, and 6-Monoacetylmorphine (6-MAM) in Urine, Blood, Serum, or Plasma Using Liquid Chromatography with Tandem Mass Spectrometry Detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 411 Tim Dahn, Josh Gunn, Scott Kriger, and Andrea R. Terrell xii Contents 41. Urine Organic Acid Analysis for Inherited Metabolic Disease Using Gas Chromatography-Mass Spectrometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 423 Patricia M. Jones and Michael J. Bennett 42. Identification of Urine Organic Acids for the Detection of Inborn Errors of Metabolism Using Urease and Gas Chromatography-Mass Spectrometry (GC-MS) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 433 Stanley F. Lo, Velta Young, and William J. Rhead 43. Quantitation of Orotic Acid in Urine Using Isotope Dilution-Selected Ion Gas Chromatography-Mass Spectrometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . 445 Jie Chen and Michael J. Bennett 44. Quantitation of Oxycodone in Blood and Urine Using Gas Chromatography-Mass Spectrometry (GC-MS) . . . . . . . . . . . . . . . . . . . . . . . . . . . 453 Bruce A. Goldberger, Chris W. Chronister, and Michele L. Merves 45. Quantitation of Phencyclidine (PCP) in Urine and Blood Using Gas Chromatography-Mass Spectrometry (GC-MS) . . . . . . . . . . . . . . . . . . . . . . . . . . . 461 Angela M. Ferguson and Uttam Garg 46. Quantitation of Sirolimus Using Liquid Chromatography-Tandem Mass Spectrometry (LC-MS-MS) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 469 Magdalena Korecka and Leslie M. Shaw 47. Quantitation of Tacrolimus in Whole Blood Using High Performance Liquid Chromatography-Tandem Mass Spectrometry (HPLC-MS-MS) . . . . . . . . . . . . . . 479 Keri J. Donaldson and Leslie M. Shaw 48. Analysis of Testosterone in Serum Using Mass Spectrometry . . . . . . . . . . . . . . . . . 489 Robert L. Fitzgerald, Terrance L. Griffin, and David A. Herold 49. Determination of Tetrahydrozoline in Urine and Blood Using Gas Chromatography-Mass Spectrometry (GC-MS) . . . . . . . . . . . . . . . . . . . . . . . . . . . 501 Judy Peat and Uttam Garg 50. Quantitation of 25-OH-Vitamin D (25OHD) Using Liquid Tandem Mass Spectrometry (LC-MS-MS) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 509 Ravinder J. Singh 51. Identification and Quantitation of Zolpidem in Biological Matrices Using Gas Chromatography-Mass Spectrometry (GC-MS) . . . . . . . . . . . . . . . . . . 519 Timothy P. Rohrig, Lydia A. Harryman, and Melissa C. Norton 52. Identification and Quantitation of Zopiclone in Biological Matrices Using Gas Chromatography-Mass Spectrometry (GC-MS) . . . . . . . . . . . . . . . . . . 527 Timothy P. Rohrig, Melissa C. Norton, and Lydia A. Harryman Subject Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 535 Contributors RYAN ALLENBRAND • Department of Pathology and Laboratory Medicine, Children’s Mercy Hospitals and Clinics, Kansas City, MO, USA SUSAN AUCOIN • Biochemical Genetics Laboratory, Department of Genetics, Children’s Hospital of Eastern Ontario, Ottawa, ON, Canada MICHAEL J. BENNETT • Department of Pathology and Laboratory Medicine, University of Pennsylvania and Children’s Hospital of Philadelphia, Philadelphia, PA, USA MATTHEW W. BJERGUM • Toxicology and Drug Monitoring Laboratory, Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, USA BRUNO CASETTA • Applied Biosystems, Monza, Italy JIE CHEN • Department of Pathology and Laboratory Medicine, The Children’s Hospital of Philadelphia, Philadelphia, PA, USA CHRIS W. CHRONISTER • Department of Pathology, Immunology, and Laboratory Med- icine, University of Florida College of Medicine, Gainesville, FL, USA WEI-LIEN CHUANG • Genzyme Corporation, Framingham, MA, USA KIM COX • Regional Forensic Science Center, Wichita, KS, USA TIM DAHN • AIT Laboratories, Indianapolis, IN, USA AMITAVA DASGUPTA • Department of Pathology and Laboratory Medicine, University of Texas-Houston Medical School, Houston, TX, USA DENNIS J. DIETZEN • Departments of Pediatrics and Pathology and Immunology, Washington University School of Medicine, St. Louis, MO, USA; St. Louis Children’s Hospital, St. Louis, MO, USA KERI J. DONALDSON • Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA, USA CAROLE S. ELBIN • Genzyme Corporation, Framingham, MA, USA CLAUDINE FASCHING • Pathology and Laboratory Medicine Service, Veterans Affairs Medical Center, Minneapolis, MN, USA ROBERT L. FITZGERALD • Veterans Affairs, San Diego Healthcare System, San Diego, CA, USA; University of California-San Diego, La Jolla, CA, USA STEVEN W. FLEMING • Department of Pathology and Laboratory Medicine, Children’s Mercy Hospitals and Clinics, Kansas City, MO, USA C. CLINTON FRAZEE III • Department of Pathology and Laboratory Medicine, Children’s Mercy Hospitals and Clinics, Kansas City, MO, USA ANGELA M. FERGUSON • Department of Pathology and Laboratory Medicine, Children’s Mercy Hospitals and Clinics, Kansas City, MO, USA MEGAN GAMBLE • Regional Forensic Science Center, Wichita, KS, USA UTTAM GARG • Department of Pathology and Laboratory Medicine, Children’s Mercy Hospitals and Clinics, Kansas City, MO, USA DIANE CIUFFETTI GEIS • Core Laboratory, McLendon Clinical Laboratories, UNC Hospitals, Chapel Hill, NC, USA xiii