Journal Published Online: 01 May 2000
Volume 45, Issue 3

Achiral and Chiral Quantification of Methamphetamine and Amphetamine in Human Urine by Semi-micro Column High-Performance Liquid Chromatography and Fluorescence Detection

CODEN: JFSCAS

Abstract

In this paper, miniaturized achiral and chiral high-performance liquid chromatographic procedures for the determination of methamphetamine and amphetamine in human urine are described. After a simple pretreatment of human urine (i.e., 10 μL of urine or diluted urine were acidified and dried-up under N2 at room temperature) and fluorescence derivatization with 4-(4,5-diphenyl-1H-imidazol-2-yl)-benzoyl chloride under mild conditions (pH 9.0, 10 min at room temperature), the derivatives were isocratically separated on a semi-micro ODS column with Tris-HCl buffer (0.1 M, pH 7.0): acetonitrile (45 + 55 v/v) at a flow rate of 0.2 mL/min or their enantiomers were separated on a semi-micro OD-RH column with sodium hexafluorophosphate (0.3 M aq.): acetonitrile (44 + 56 v/v) at a flow rate of 0.1 mL/min as the mobile phase. Wide-ranged calibration curves were obtained with detection limits for the achiral and chiral analyses in the atto and femtomol levels, respectively, per injected volume. Satisfactory within- and between-day reproducibility data were obtained with both the methods with the highest relative standard deviation being 9.6%. The methods were applied to the determination of methamphetamine and amphetamine in human urine samples and the concentrations determined by the two methods were well correlated (r = 0.994).

Author Information

Al-Dirbashi, OY
Graduate School of Pharmaceutical Sciences, Nagasaki University, Nagasaki, Japan
Wada, M
School of Pharmaceutical Sciences, Nagasaki University, Nagasaki, Japan
Kuroda, N
School of Pharmaceutical Sciences, Nagasaki University, Nagasaki, Japan
Takahashi, M
Graduate School of Pharmaceutical Sciences, Nagasaki University, Nagasaki, Japan
Nakashima, K
Graduate School of Pharmaceutical Sciences, Nagasaki University, Nagasaki, Japan
Pages: 7
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Stock #: JFS14755J
ISSN: 0022-1198
DOI: 10.1520/JFS14755J