DEVELOPMENT OF LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY (LC-MS/MS) METHODS FOR EXPLORATION OF CURRENCY CONTAMINATION WITH CONTROLLED SUBSTANCES By ALLISON M. VEITENHEIMER Bachelor of Science in Forensic Science Baylor University Waco, Texas 2008 Submitted to the Faculty of the Graduate College of the Oklahoma State University in partial fulfillment of the requirements for the Degree of MASTER OF SCIENCE July 2010 DEVELOPMENT OF LIQUID CHROMATOGRAPHY- TANDEM MASS SPECTROMETRY (LC-MS/MS) METHODS FOR EXPLORATION OF CURRENCY CONTAMINATION WITH CONTROLLED SUBSTANCES Thesis Approved: Dr. Jarrad R. Wagner Thesis Adviser Dr. Robert W. Allen Committee Member Dr. David R. Wallace Committee Member Dr. Mark E. Payton Dean of the Graduate College ii ACKNOWLEDGMENTS First and foremost, I would like to thank my advisor Dr. Jarrad Wagner. His encouragement, guidance, and support throughout the entirety of this project helped me tremendously. I would also like to thank my committee members, Dr. Robert Allen and Dr. David Wallace. Their comments and suggestions were greatly appreciated. The faculty and staff in the Forensic Sciences Department have also provided great support throughout this process. A special thanks to Dr. Thomas Jourdan for allowing me to assist him with this project. This entire experience, up to and including a trip to the 62nd Annual Scientific Meeting of the American Academy of Forensic Sciences to present this data, was made possible due to his ongoing research. I am deeply grateful to my family and friends for their continued support over the years. Their belief in me has meant more to me than they know. I would like to dedicate this thesis to my parents, James and Lisa Veitenheimer, without whom I would not be where I am today. Their unconditional love and support has allowed me to achieve things I never thought possible and for that I thank them. iii TABLE OF CONTENTS Topic Page Chapter I. Introduction.........................................................................................................1 Chapter II. Review of Literature..........................................................................................4 2.1. Currency Contamination ...........................................................................4 2.1.1. Extraction Methods..........................................................................7 2.1.1.1. Vacuum Sampling..................................................................8 2.1.1.2. Thermal Desorption................................................................8 2.1.1.3. Solvent Extraction..................................................................9 2.1.1.3.1. Solid Phase Extraction ................................................10 2.1.2. Analytical Methods........................................................................10 2.1.2.1. Immunoassay........................................................................11 2.1.2.2. Thermal Desorption Tandem Mass Spectrometry ...............11 2.1.2.3. Ion Mobility Spectrometry...................................................11 2.1.2.4. Gas Chromatography with Mass Spectrometry ...................12 2.1.2.5. Liquid Chromatography with Mass Spectrometry ...............13 2.1.2.6. Capillary Electrophoresis .....................................................13 2.1.2.7. Desorption Electrospray Ionization (DESI) with Mass Spectrometry ...........................................................................................13 2.1.2.8. Raman Spectroscopy ............................................................14 2.1.3. Cocaine Contamination Studies.....................................................14 2.1.4. Illicit Drug Contamination Studies................................................16 iv 2.2. Specific Drugs.........................................................................................19 2.2.1. Cocaine ..........................................................................................20 2.2.2. Codeine..........................................................................................22 2.2.3. Heroin ............................................................................................22 2.2.4. MDMA ..........................................................................................23 2.2.5. Methamphetamine .........................................................................24 2.2.6. Morphine........................................................................................25 2.2.7. THC ...............................................................................................25 2.3. Liquid Chromatography with Tandem Mass Spectrometry....................26 2.3.1. Liquid Chromatography.................................................................26 2.3.2. Mass Spectrometry ........................................................................27 2.3.2.1. Ionization..............................................................................28 2.3.2.2. Ion Separation ......................................................................31 2.3.2.3. Ion Detection ........................................................................33 2.4. Method Development..............................................................................34 2.5. Method Validation...................................................................................36 2.5.1. Accuracy........................................................................................36 2.5.2. Precision ........................................................................................37 2.5.3. Selectivity ......................................................................................37 2.5.3.1. Matrix Effects.......................................................................38 2.5.4. Sensitivity ......................................................................................41 2.5.5. Reproducibility and Stability.........................................................41 v 2.5.6. Linearity.........................................................................................42 2.5.7. Carryover .......................................................................................42 Chapter III. Methodology ..................................................................................................44 3.1. Instrumentation........................................................................................44 3.2. Materials..................................................................................................45 3.3. Preparation of Standards .........................................................................46 3.4. Method Development..............................................................................47 3.4.1. Cocaine Method.............................................................................47 3.4.1.1. LC Parameters ......................................................................47 3.4.1.2. Source Parameters ................................................................48 3.4.1.3. MS Parameters .....................................................................48 3.4.2. Multi Drug Method........................................................................49 3.4.2.1. LC Parameters ......................................................................50 3.4.2.2. Source Parameters ................................................................50 3.4.2.3. MS Parameters .....................................................................50 3.5. Sample Collection ...................................................................................52 3.6. Preparation of Samples............................................................................53 3.7. Analytical Procedure ...............................................................................53 3.8. Method Validation...................................................................................54 3.8.1. Precision ........................................................................................54 3.8.2. Accuracy........................................................................................55 3.8.3. Sensitivity ......................................................................................55 vi 3.8.4. Selectivity ......................................................................................55 3.9. Quantitation of Currency Contamination................................................56 3.10. Statistical Analysis ..................................................................................57 Chapter IV. Results............................................................................................................58 4.1. Precision..................................................................................................58 4.1.1. Cocaine Method.............................................................................58 4.1.2. Multi Drug Method........................................................................58 4.2. Accuracy..................................................................................................59 4.2.1. Cocaine Method.............................................................................59 4.2.2. Multi Drug Method........................................................................59 4.3. Sensitivity................................................................................................60 4.3.1. Cocaine Method.............................................................................60