Simple and sensitive high-performance liquid chromatographic method for the determination of 1,5-benzodiazepine clobazam and its active metabolite N-desmethylclobazam in human serum and urine with application to 1,4-benzodiazepines analysis.
A HPLC-UV determination of clobazam and N-desmethylclobazam in human serum and urine is presented. After simple liquid-liquid extraction with dichloromethane the compounds and an internal standard diazepam were separated on a Supelcosil LC-8-DB column at ambient temperature under isocratic conditions using the mobile phase: CH3CN-water-0.5 M KH2PO4-H3PO4 (440:540:20:0.4, v/v and 360:580:60:0.4, v/v for serum and urine, respectively). The detection was performed at 228 nm with limits of quantification of 2 ng/ml for serum and 1 ng/ml for urine. Relative standard deviations for intra- and inter-assay precision were found below 8% for both compounds for all the tested concentrations. The described procedure may be easily adapted for several 1,4-benzodiazepines.
Kunicki
P K
PK
Clinical Pharmacology Unit, Department of Clinical Biochemistry, National Institute of Cardiology, Warsaw, Poland. biblnauk@ikard.waw.pl
D014151:Anti-Anxiety Agents / Q000097:blood / Q000493:pharmacokinetics* / Q000652:urine; D001569:Benzodiazepines / Q000097:blood / Q000493:pharmacokinetics* / Q000652:urine; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D000078306:Clobazam; D006801:Humans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D014151:Anti-Anxiety Agents; D001569:Benzodiazepines; D000078306:Clobazam; C024581:N-desmethylclobazam