Residue from smoked methamphetamine hydrochloride contains pyrolytic products that are detectable by gas chromatography-mass spectrometry (GC-MS). A validated GC-MS method was developed for the determination of trans-phenylpropene, a pyrolytic product of methamphetamine HCl, in residue of smoked drug as well as in human urine. trans-Phenylpropene and an isomeric internal standard, 2-phenylpropene, were extracted from urine using n-hexane. The method was validated for linearity over a range of 0.1-10 microg/mL with a limit of detection of 0.05 microg/mL, limit of quantification of 0.1 microg/mL, interday accuracy within 10.5%, intraday accuracy better than 3.7%, interday precision of 15.4%, intraday precision of 14.4%, and recovery of 89.1%. The method was applied to the detection of trans-phenylpropene found in the residue of methamphetamine HCl heated beyond its melting temperature on aluminum foil under simulated smoking conditions. The method is applicable to the detection of trans-phenylpropene in urine as a potential marker for smoked methamphetamine HCl abuse.

Metallothioneins (MT) are cysteine-rich intracellular proteins that sequester epoxides, such as the genotoxic styrene metabolite, styrene oxide (SO). Covalent adduct formation by the ring-opening of SO and its homologue, 1-phenylpropylene oxide (trans), with MT forms stable complexes that potentially may serve as biomarkers for epoxide exposure. MT was reacted with SO and trans-phenylpropylene oxide (PPO) separately in water. Mass spectra of adducts were obtained using electrospray ionization with positive-ion detection in an ion trap mass spectrometer. MS/MS analysis established the covalent nature of the complexation. Further experiments were conducted to determine the site of adduct formation. Molecular modeling was used to predict the most likely site of adduct formation. Alkylation and digestion were used to locate the peptide fragment where adduct formation occurred and confirm modeling results. The likeliest cysteine residue was residue 48 in the alpha domain of MT. MT-SO adducts provide a basis for the development of biomarkers. iii ACKNOWLEDGMENTS I would first like to thank my advisor, Dr. Patrick Callery, for his advice and confidence in me. He has been tremendous help to me in this whole process. I would also like to thank Dr. Grazyna Szklarz and Dr. Fred King for agreeing to serve on my committee. I want to also thank Dr. Diaa Shakleya for all of his help and guidance throughout this project. I want to also thank the entire Pharmaceutical Sciences Department, faculty, staff, and students for all of their help and support. I wish to thank Mylan Pharmaceuticals and my colleagues in the Bioanalytical Department, especially Dr. Scott Chervenick, who gave me the time and encouragement to continue my education. Finally, I would also like to especially thank my mother, family, and friends for all of their love, encouragement, and support. iv TABLE OF CONTENTS Abstract ii Acknowledgments iii Table of Contents iv List of Figures v List of Tables vi List of abbreviations vii