SOFT 2004 Meeting Abstracts - Society of Forensic Toxicologists

M16
VALIDA TION OF THE COZAR~
AMPHETAMINES IN HAIR
MICROPLATE ELISA FOR THE DETECTION OF
Gail Cooper* and Chris Hand: Cozart Bioscience Ltd, Abingdon, Oxfordshire, OXI4 4RU
The purpose of this study was to determine the performance characteristics of the Cozart® Amphetamine
Microplate ELISA assay for the detection of amphetamines in hair samples. Hair samples (N=50) were
collected from volunteers with a history of drug use and from drug related deaths. The hair samples were
sonicated in methanol and then extracted overnight at 60°C. The methanol extract was evaporated to
dryness and reconstituted in ELISA negative calibrator. 251lL of calibrator (0, 50, 100,200,300,500 and
1000pg/mg), control and sample were assayed in duplicate according to manufacturers instructions for the
Cozart@ Amphetamine ELISA - Forensic Application Kit.
For GC-MS analysis, deuterated internal standards (amphetamine-d5, methamphetamine-d5, MDA-d5,
MDMA-d5, MDEA-d6 and MBDB-d5) mixture, p-glucuronidase and O.lM, pH 6.0 phosphate buffer were
added to approximately 20 mg of sample, or spiked blank hair and sonicated for I hour. Isolation of
amphetamine, methamphetamine, MDA, MDMA, MDEA and MBDB was achieved through simple solidphase
extraction using Bond Elut certifY cartridges.
The true positives (N=20), true negatives (N=30), false positives (N=O) and false negatives (N=O) for
different cut-offs with the ELISA were determined by comparison of the ELISA response (normalised to
weight of hair extracted) to the GC-MS results as the reference method. Hair concentrations ranged from O.
- 10.1 ng/mg (amphetamine), 0 - 0.8 ng/mg (MDA) and 0 17.4 ng/mg (MDMA).
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The optimum cut-off for the Cozart® Amphetamine Microplate ELISA was determined to be 500pg
amphetamine equivalents/mg hair using a 20 mg hair sample with an overall precision of 100% vs GC-MS.
Keywords: Amphetamines, Hair, ELISA
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