SIMULTANEOUS HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC ANALYSIS OF FLUNITRAZEPAM AND 4 METABOLITES IN SERUM

A high-performance liquid chromatographic method for the simultaneous determination of flunitrazepam and four metabolites, desmethylflunitra zepam (DMF), 7-aminodesmethylflunitrazepam (7-NH(2)DMF), 7-aminoflunit razepam (7-NH2F) and 3-hydroxyflunitrazepam (3-OHF), in serum is descr ibed. The method involves a simple extraction from alkalinized plasma (pH 9.5) into diethyl ether-chloroform (80:20, v/v). Prazepam was used as an internal standard for the quantification of the five compounds. Separation was achieved with a 10 mu m RSil CN column (300x3.9 mm I.D .). The detection wavelength was set at 242 nm. The limits of detectio n ranged from 2.5 to 5 mu g/l with a limit of quantification of 10 mu g/l for all analytes.