Simultaneous Determination of Phencyclidine and Its Metabolites in Rat Urine by High-Performance Liquid Chromatography

Abstract

A high-performance liquid chromatography with chemiluminescence detection using tris(2,2′-bipyridyl)ruthenium(II) [Ru(bpy)₃²⁺] for the simultaneous determination of phencyclidine and its major metabolites in rat urine was developed. The urine sample was applied onto a disposable cartridge column (Bond Elut Certify) of silica solid-phase extraction. The charged sample was then washed with an acidic solution and methanol prior to its elution with 2% ammonium hydroxide in methanol, and the eluate was chromatographed. In this method, a TSKgel ODS 80Ts column and a mobile phase containing 60% methanol, 50 mM phosphate buffer (pH 7.0) and 0.12 mM tris(2,2′- bipyridyl)chloride hexahydrate in 10 mM sulfuric acid as a reagent solution were used. The calibration curves for phencyclidine and its metabolites showed good linearity over the range 25 - 400 ng ml^-1 per 20 μl injection. As a result, the method was successfully applied to a simultaneous determination of phencyclidine and its three major metabolites in actual rat urine.