High-Pressure Liquid Chromatographic Analysis of Triflubazam and Its Metabolites in Human and Animal Blood and Urine

A high-pressure liquid chromatographic method is described for analyzing triflubazam [1-methyl-5-phenyl-7-trifluoromethyl-1H-1,5-benzodiazepine-2,4(3H,5H)-dione] and its primary metabolites in blood and urine. Adsorption chromatography, using pellicular silica gel as the stationary phase and dioxane-isooctane as the mobile phase, permitted rapid sample analysis. After extraction of blood and urine samples with toluene, quantitation is achieved using liquid chromatography with an internal standard. The method is sensitive above 50 ng/ml of triflubazam and its known metabolites. Recoveries for all compounds from blood or urine averaged above 95%. The specificity of the method was established by collecting samples separated by liquid chromatography and characterizing them by mass spectrometry. Human and animal data are presented to illustrate the utility of the method.