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  • 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.

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.

Journal of chromatography. B, Biomedical sciences and applications (2001-02-24)
P K Kunicki
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Supelco
SUPELCOSIL LC-8-DB Supelguard Cartridge, 5 μm particle size, L × I.D. 2 cm × 4 mm
Supelco
SUPELCOSIL LC-8-DB HPLC Column, 5 μm particle size, L × I.D. 25 cm × 4 mm
Supelco
SUPELCOSIL LC-8-DB HPLC Column, 5 μm particle size, L × I.D. 15 cm × 4.6 mm
Supelco
SUPELCOSIL LC-8-DB Supelguard Cartridge, 5 μm particle size, L × I.D. 2 cm × 4 mm
Supelco
SUPELCOSIL LC-8-DB HPLC Column, 5 μm particle size, L × I.D. 25 cm × 4.6 mm
Supelco
SUPELCOSIL LC-8-DB HPLC Column, 5 μm particle size, L × I.D. 5 cm × 4.6 mm
Supelco
SUPELCOSIL LC-8-DB Supelguard Cartridge, 5 μm particle size, L × I.D. 2 cm × 3 mm
Supelco
SUPELCOSIL LC-8-DB HPLC Column, 3 μm particle size, L × I.D. 3.3 cm × 2.1 mm
Supelco
SUPELCOSIL LC-8 HPLC Column, 5 μm particle size, L × I.D. 25 cm × 10 mm
Supelco
SUPELCOSIL LC-8 HPLC Column, 3 μm particle size, L × I.D. 3.3 cm × 4.6 mm
Supelco
SUPELCOSIL LC-8 HPLC Column, 5 μm particle size, L × I.D. 5 cm × 4.6 mm
Supelco
SUPELCOSIL LC-8 HPLC Column, 3 μm particle size, L × I.D. 15 cm × 4.6 mm
Supelco
SUPELCOSIL LC-8 HPLC Column, 5 μm particle size, L × I.D. 25 cm × 4.6 mm
Supelco
SUPELCOSIL LC-8 HPLC Column, 5 μm particle size, L × I.D. 15 cm × 4 mm
Supelco
SUPELCOSIL LC-NH2-NP HPLC Column, 5 μm particle size, L × I.D. 25 cm × 4.6 mm
Supelco
SUPELCOSIL LC-NH2-NP Supelguard Cartridge, 5 μm particle size, L × I.D. 2 cm × 4 mm
Supelco
SUPELCOSIL LC-8 Supelguard Cartridge, 5 μm particle size, L × I.D. 2 cm × 4 mm
Supelco
SUPELCOSIL LC-8 Supelguard Cartridge, 5 μm particle size, L × I.D. 2 cm × 4 mm
Supelco
SUPELCOSIL LC-8 HPLC Column, 5 μm particle size, L × I.D. 15 cm × 4.6 mm
Supelco
SUPELCOSIL LC-8-DB HPLC Column, 3 μm particle size, L × I.D. 3.3 cm × 4.6 mm
Supelco
SUPELCOSIL LC-8 Supelguard Cartridge, 5 μm particle size, L × I.D. 2 cm × 2.1 mm
Supelco
SUPELCOSIL LC-8-DB HPLC Column, 3 μm particle size, L × I.D. 15 cm × 4.6 mm
Supelco
SUPELCOSIL LC-8 HPLC Column, 3 μm particle size, L × I.D. 7.5 cm × 4.6 mm
Supelco
N-Desmethylclobazam solution, 100 μg/mL in acetonitrile, ampule of 1 mL, certified reference material, Cerilliant®