Analytical Chemistry in Medical Science and Technology. Determination of methamphethamine by HPLC with chemiluminescence detection.

A method is described for the determination of methamphetamine (MP) by a high-performance liquid-chromatographic method using a peroxyoxalate chemiluminescence reaction. After MP was derivatized with 4-(N, N-dimethylaminosulphony1)-7-fluoro-2, 1, 3 benzoxadiazole (DBD-F), it was detected by a chemiluminescence method. Bis(2, 4, 6 trichlorophenyl) oxalate (TCPO) and hydrogen peroxide were used as postcolumn chemiluminescence reagents. The separation of the DBD-F derivative of MP was carried out using a reversed-phase column (GL Science, Lichrospher RP 18-5, 4.6 i.d. × 250 mm) and an acetonitrile aqueous solution as the mobile phase(flow rate, 1.0 ml/min). The optimum pH, temperature and reaction time for fluorogenic reactions of DBD-F and MP were 8, 70°C, 30 min, respectively. The linear calibration graph was observed over the range of 5 × 10-95 ×10-6 M. The correlation coefficient for this line was 0.999. The detection limit (3σ) was 2× 10-9 M (40 fmol) and the relative standard deviation was 1.4% (n=6) for a concentration of 5 × 10-9 M. The effect of urine on the MP determination was investigated. The urine sample solutions were prepared by adding a 1 × 10-7 M or 1 × 10-8 M MP standard solution to a normal human urine. The relative chemiluminescence intensities were 98102% toward the chemiluminescence intensity of the MP standard solution.