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A high performance liquid chromatographic method for the determination of an agent of benzodiazepine receptor inverse agonist (S-135) was developed. S-135 in monkey plasma, after its administration, was assayed using the method. S-135 is ionized in alkaline N,N-dimethylformamide (DMF), and fluoresces strongly at 490 nm on excitation at 290 nm. Extracted S-135 from a plasma sample was separated with an Asahipak GS-310H polyvinyl alcohol gel column and detected fluorometrically, with the eluate being mixed with a triethylamine-DMF mixture. S-135 in plasma is assayable with a coefficient of variation being less than 3.5% (n = 11). The lower limit of the determination is 2 ng/ml. This method was applied as a highly sensitive and simple method to pharmacokinetic studies of S-135 after oral administration to cynomolgus monkeys.

Fluorometric Determination of an Inverse Agonist to Benzodiazepine Receptors S-135 in Monkey Plasma by High Performance Liquid Chromatography.