Published: Journal of Analytical Toxicology, ISSN 0146-4760, Volume 31, Number 4, May 2007, pp. 208-214

Matrix Effect and Cross-Reactivity of Select Amphetamine-Type Substances, Designer Analogues, and Putrefactive Amines using the Bio-Quant Direct ELISA Presumptive Assays for Amphetamine and Methamphetamine
Luigino G. Apollonio[1], Ian R. Whittall[2], Dennis J. Pianca[2], Jennelle M. Kyd[1], and William A. Maher[1]
[1]National Centre for Forensic Studies, University of Canberra, Bruce ACT 2601 Australia and
[2]Department of Toxicology and Forensic Chemistry, ACT Government Analytical Laboratory, Weston Creek ACT 2611 Australia

The aim of this study was to evaluate the Bio-Quant Direct ELISA assays for amphetamine and methamphetamine in the routine presumptive screening of biological fluids. Standard concentration curves of the target analytes were assayed to assess sensitivity, and known concentrations of common amphetamine-type substances (ephedrine, pseudoephedrine, phentermine), designer analogues (MDA, MDMA, MDEA, MBDB, PMA, 4-MTA, 2CB), and putrefactive amines (phenylethylamine, putrescine, tryptamine, tyramine) were analyzed to determine cross-reactivity. Results of the standard curve studies show the capacity of both Direct ELISA kits to confidently detect down to 3 ng/mL interday (PBS matrix; CVs 6.3–15.5%). Cross-reactivity relative to that of 50 ng/mL preparations of the target compounds demonstrated that the Direct ELISA kit for amphetamine also detected MDA (282%), PMA (265%), 4-MTA (280%), and phentermine (61%), and the Direct ELISA for methamphetamine also assayed positive for MDMA (73%), MDEA (18%), pseudoephedrine (19%), MBDB (8%), and ephedrine (9%). Matrix studies demonstrated that both ELISA kits could be applied to screening of blood, urine, and saliva to a concentration of 6 ng/mL or lower. In conclusion, the Bio-Quant Direct ELISA kits for amphetamine and methamphetamine are fast and accurate and have demonstrated themselves to be useful tools in routine toxicological testing.

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