Simultaneous determination of clobazam and its major metabolite in human plasma by a rapid HPLC method

Rouini, M.; Ardakani, Y.H.; Hakemi, L.; Mokhberi, M.; Badri, G.

Journal of Chromatography. B Analytical Technologies in the Biomedical and Life Sciences 823(2): 167-171


ISSN/ISBN: 1570-0232
PMID: 16005690
DOI: 10.1016/j.jchromb.2005.06.031
Accession: 012559464

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A rapid and specific HPLC method has been developed and validated for simultaneous determination of clobazam, the anticonvulsant agent, and its major metabolite in human plasma. The sample preparation was a liquid-liquid extraction with tuloene yielding almost near 100% recoveries of two compounds. Chromatographic separation was achieved with a Chromolith Performance RP-18e 100 mm x 4.6mm column, using a mixture of a phosphate buffer (pH 3.5; 10mM)-acetonitrile (70:30, v/v), in isocratic mode at 2 ml/min at a detection wave-length of 228 nm. The calibration curves were linear (r(2)>0.998) in the concentration range of 5-450 ng ml(-1). The lower limit of quantification was 5 ng ml(-1) for two compounds studied. The within- and between-day precisions in the measurement of QC samples at four tested concentrations were in the range of 0.89-9.1% and 2.1-10.1% R.S.D., respectively. The developed procedure was applied to assess the pharmacokinetics of clobazam and its major metabolite following administration of a single 10mg oral dose of clobazam to healthy volunteers.