Simultaneous Determination of Clobazam, N-Desmethyl Clobazam and Clonazepam in Plasma by High Performance Liquid Chromatography

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Abstract
A method is described for the simultaneous determination of clobazam, its active metabolite /V-desmethylclobazam, and clonazepam in plasma by high performance liquid chromatography. The drugs are extracted with diethyl ether from 1.0 ml plasma adjusted to pH 9.0 and separated on a C18 reverse phase column using a mobile phase of 40% acetonitrile in phosphate buffer pH 3.0. The ultraviolet absorbance is monitored at a wavelength of 313 nm. The method has a coefficient of variation of < 5% and an overall recovery of > 85% and is sufficiently sensitive for the therapeutic monitoring of these drugs.