Simultaneous Detection and Quantification of Amphetamines, Diazepam and its Metabolites, Cocaine and its Metabolites, and Opiates in Hair by LC-ESI-MS-MS Using a Single Extraction Method

• Published in: Journal of analytical toxicology 2008-09, Vol.32 (7), p.457-469
• Main Authors: Miller, Eleanor I., Wylie, Fiona M., Oliver, John S.
• Format: Article
• Language: English
• Subjects: Amphetamines - analysis; Amphetamines - poisoning; Analgesics, Opioid - analysis; Analgesics, Opioid - poisoning; Analysis; Biological and medical sciences; Buffers; Chromatography, High Pressure Liquid; Cocaine - analysis; Cocaine - poisoning; Diazepam - analysis; Diazepam - poisoning; Drug addictions; Drug Overdose - diagnosis; Drug Overdose - mortality; General pharmacology; Hair - chemistry; Humans; Indicators and Reagents; Medical sciences; Methanol - chemistry; Morphine Derivatives - analysis; Pharmacology. Drug treatments; Phosphates - chemistry; Reference Standards; Reproducibility of Results; Solid Phase Extraction; Solvents; Spectrometry, Mass, Electrospray Ionization; Substance Abuse Detection - methods; Toxicology
• ISSN: 0146-4760; 1945-2403
• DOI: 10.1093/jat/32.7.457

A liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous identification and quantification of amphetamines, diazepam and its metabolites, cocaine and its metabolites, and opiates from hair using a single extraction method. As part of the method development, Gemini C18, Synergi Hydro RP, and Zorbax Stablebond-Phenyl LC columns were tested with three different mobile phases. Analyte recovery and limit of detection were evaluated for two different solid-phase extraction methods that used Bond Elut Certify™ and Clean Screen® cartridges. Phosphate buffer (pH 5.0) was chosen as the optimum hair incubation medium because of the high stability of cocaine and 6-monoacetylmorphine using this method and faster sample preparation. The optimized method was fully validated. Linearity was established over the concentration range 0.2–10 ng/mg hair, and the correlation coefficients were all greater than 0.99. Total extraction recoveries were greater than 76%, detection limits were between 0.02 and 0.09 ng/mg, and the intra- and interday imprecisions were generally less than 20% in spiked hair. The intra- and interbatch imprecision of the method for a pooled authentic hair sample ranged from 1.4 to 23.4% relative standard deviation (RSD) and 8.3 to 25.4% RSD, respectively, for representative analytes from the different drug groups. The percent matrix effect ranged from 63.5 to 135.6%, with most analytes demonstrating ion suppression. Sixteen postmortem samples collected from suspected drug-related deaths were analyzed for the 17 drugs of abuse and metabolites included in the method. The method was sufficiently sensitive and specific for the analysis of drugs and metabolites in postmortem hair samples. There is scope for the inclusion of other target drugs and metabolites in the method.