Determination of the anxiolytic agent 8-chloro-6-(2-chlorophenyl)-4H-imidazo-[1,5-alpha] [1,4]-benzodiazepine-3-carboxamide in whole blood, plasma or urine by high-performance liquid chromatography.
A rapid, sensitive and specific high-performance liquid chromatographic (HPLC) assay was developed for the determination of 8-chloro-6-(2-chlorophenyl)-4H-imidazo-[1,5-alpha]-[1,4]-benzodiazepine-3-carboxamide [I] and its 4-hydroxy metabolite, 8-chloro-6-(2-chlorophenyl)-4-hydroxy-4H-imidazo-[1,5-alpha] [1,4]-benzodiazepine-3-carboxamide [II] in whole blood, plasma or urine. The assay for both compounds involves extraction into diethyl ether-methylene chloride (70:30) from blood, plasma, or urine buffered to pH 9.0. The overall recoveries of [I] and [II] are 92.0 +/- 5.4% (S.D.) and 90.3 +/- 4.9% (S.D.), respectively. The sensitivity limit of detection is 50 ng/ml of blood, plasma, or urine using a UV detector at 254 nm. The HPLC assay was used to monitor the blood concentration-time fall-off profiles, and urinary excretion profiles in the dog following single 1 mg/kg intravenous and 5 mg/kg oral doses, and following multiple oral doses of 100 mg/kg/day of compound [I].
D000818:Animals; D014151:Anti-Anxiety Agents / Q000008:administration & dosage / Q000097:blood* / Q000652:urine; D001569:Benzodiazepines / Q000008:administration & dosage / Q000097:blood* / Q000652:urine; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004285:Dogs; D007275:Injections, Intravenous; D008874:Midazolam / Q000031:analogs & derivatives*; D012016:Reference Values; D013997:Time Factors
D014151:Anti-Anxiety Agents; D001569:Benzodiazepines; C033415:8-chloro-6-(2-chlorophenyl)-4H-imidazo(1,5-a)(1,4)-benzodiazepine-3-carboxamide; D008874:Midazolam