Achiral and chiral quantification of methamphetamine and amphetamine in human urine by semi-micro column high-performance liquid chromatography and fluorescence detection.
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 microL 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).
Al-Dirbashi
O Y
OY
Department of Analytical Research for Pharmacoinformatics, Graduate School of Pharmaceutical Sciences, Nagasaki University, Japan.
Wada
M
M
Kuroda
N
N
Takahashi
M
M
Nakashima
K
K
D000661:Amphetamine / Q000652:urine*; D000697:Central Nervous System Stimulants / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000592:standards; D005453:Fluorescence; D005554:Forensic Medicine / Q000379:methods; D006801:Humans; D008694:Methamphetamine / Q000652:urine*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000697:Central Nervous System Stimulants; D008694:Methamphetamine; D000661:Amphetamine