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DEVELOPMENT AND VALIDATION OF A RP-HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF PROPRANOLOL HCl AND CLONAZEPAM IN BULK AND PHARMACEUTICAL DOSAGE FORM

Keywords: Propranolol HCl , Clonazepam , RP-HPLC , Simultaneous determination , Validation.

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Abstract:

The present work describes a reverse phase high performance liquid chromatographic method (RP-HPLC) for the simultaneous estimation of Propranolol HCl (PRH) and Clonazepam (CNZ) in bulk and in pharmaceutical dosage form. Chromatographic separation was performed on Agilent Eclipse xdb C18 (150 mm 4.6 mm i.d., 5 m) column, with a mobile phase comprising of a mixture of methanol, acetonitrile and 20 mM potassium dihydrogen phosphate buffer in the ratio of 27.5:27.5:45 v/v. The pH of buffer was adjusted to 3.0 with orthophosphoric acid. The flow rate was 1.0 ml/min with detection at 266 nm. Retention times of Propranolol HCl and Clonazepam were found to be 2.400 and 4.492 min respectively. As per International Conference on Harmonisation (ICH) guidelines the method was validated for linearity, accuracy, precision, limit of quantitation, limit of detection, and robustness. Linearity of PRH was found to be in the range of 20-120 μg/mL and that for CNZ was found to be 1-6 μg/mL. The correlation coefficients were 0.9994 and 0.9995 for PRH and CNZ respectively. The mean recoveries obtained for PRH and CNZ were 100.6% and 100.1%. This demonstrates that the developed method is simple, precise, accurate, reproducible and rapid for simultaneous estimation of these drugs in bulk and in tablet dosage forms.

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