Abstract
A method for the determination of amphetamine and related compounds in urine based on on-line derivatization with 9-fluorenylmethyl chloroformate (FMOC) and high-performance liquid chromatography is described. Derivatization is performed in a 20×2.1 mm I.D. column packed with a Hypersil ODS C 18, 30 μm stationary phase, which is also used for sample clean-up and enrichment of the analytes. Next, the derivatized analytes are transferred to a LiChrospher 100 RP-C 18 (5 μm, 125×4 mm I.D.) analytical column for their separation and quantification, using reversed-phase conditions and fluoresccence detection. The described assay was applied to the determination of norephedrine, ephedrine, pseudoephedrine, amphetamine, phenylpropylamine and methamphetamine at concentrations of 0.5–10.0 μg/ml. Analyte conversions were about 55–96% of those obtained by the off-line derivatization mode under similar conditions, resulting in limits of detection in the 5–25 ng/ml range.
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