Development and Validation of a RP-HPLC-PDA Method for Simultaneous Determination of Phenobarbital and Phenytoin in Pure Form and its Pharmaceutical Dosage Form
Abstract
Aim: A rapid and precise Reverse Phase High Performance Liquid chromatographic method has been developed and validated for Phenobarbital and Phenytoin, in its pure form as well as in combined tablet dosage form.
Study Design: Analytical method development and validation study.
Methodology: Chromatography was carried out on a Symmetry C18 (4.6 x 150mm, 5µm) column using a mixture of Methanol: Phosphate Buffer pH 3.5 (65:35) as the mobile phase at a flow rate of 1.0ml/min, the detection was carried out at 270 nm.
Results: The retention time of the Phenobarbital and Phenytoin was 2.459, 4.322 min respectively. The method produces linear responses in the concentration range of 5-25mg/ml of Phenobarbital and 2.5-12.5mg/ml of Phenytoin. The method’s precision for the determination of assay was below 2.0%RSD.
Conclusion: All the system suitability parameters and validation parameters also met the ICH validation acceptance criteria. Hence, the present developed method can be successfully applicable for routine quality control analysis of bulk and pharmaceutical formulations.
Keywords: Phenobarbital, Phenytoin, RP-HPLC, Method development and Validation.
Keywords:
Phenobarbital, Phenytoin, RP-HPLC, Method development and ValidationDOI
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