A RAPID AND SENSITIVE LC-MS/MS ASSAY FOR THE DETERMINATION OF CLOBAZAM IN HUMAN PLASMA USING ELECTRO SPRAY IONIZATION TECHNOLOGY

A simple, highly sensitive, precise and accurate liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed for the rapid quantification of clobazam in human plasma samples. The chromatographic separation was achieved with a reverse phase column Reprosil Gold XBD C18 (100 x 3mm, 3 mu m) and the mobile phase consisted of 10 mM ammonium acetate buffer and 0.1% acetic acid in methanol (15:85, v/v) as eluent by running a linear gradient method. The effluence was ionized by positive electrospray ionization and measured by mass spectrometry. The method was validated with linear calibration curve over a wide dynamic concentration range of 0.501-499.995 ng/mL and the lower limit of quantification was 0.501 ng/mL with good accuracy and precision. The mean extraction recovery of the method was higher than 96.4% and 103.3% for clobazam and IS, respectively. The method was successfully demonstrated for evaluation of bioequivalence study of 10 mg clobazam tablet formulation in 28 healthy Indian male subjects under fasting condition.