ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY ANALYSIS OF FLUNITRAZEPAM AND 7-AMINOFLUNITRAZEPAM IN HUMAN PLASMA

摘要

A high-throughput ultra-performance liquid chromatography (UPLC) method with UVdetection was elaborated and validated for the simultaneous quantification of Jlunitrazepam and its major metabolite, 7-aminojlunilrazepam, in human plasma. After a simple liquid-liquid extraction with ethyl acetate, chromatographic separation was performed on a BEH C18 chromatographic column with a binary mixture of acelonitrile/10 mM potassium monohydrogen phosphate (pH= 8.5) as the mobile phase, under gradient elution (flow rate= 0.275 mL/min). The total run time of the chromatographic analysis was 4.2 min. UV detection was performed at 314nm for Jlunitrazepam and at 242 nm for 7-aminojlunitrazepam. To our knowledge, this is the first reported UPLC-PDA method for the quantification of Jlunitrazepam and 7-aminojlunilrazepam in human plasma. The assay was applied successfully for the analysis of the selected two analytes in real human plasma samples obtained from a healthy volunteer, after ingeslion of a single dose of Jlunitrazepam.