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RP-HPLC Method Development and Validation for Estimation of Glibenclamide in Tablet Dosage Form
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The prime aim of the current work is to develop and validate a novel, sensitive, reverse phase High Performance Liquid Chromatography (RP-HPLC) technique for the estimation of Glibenclamide in dosage form. Chromatographic separation was achieved on a Chromosil column, (150mm×4.6mm x5μ) using an isocratic method with mobile phase composed of Potassium di-hydrogen phosphate buffer (pH 4.5): Acetonitrile in the ratio 60:40 v/v. The flow rate was 1 ml/min, temperature of the column was maintained at ambient and detection was made at 233 nm. The run time was 12 min. The developed method was validated according to the International Conference on Harmonization (ICH) guidelines with respect to linearity, accuracy, precision, specificity and robustness. The developed method was linear for Glibenclamide from 10 - 50 μg/ml and the linear regression obtained was > 0.999. Precision, evaluated by intraand inter-day assays had relative standard deviation (R.S.D) values within 1.5%. Recovery data were in the range 98.2% to 100.9% with R.S.D. values < 1.5 %. The method is precise, accurate, linear, robust and fast. The short retention time allows the analysis of a large number of samples in a short period of time and, therefore, should be costeffective for routine Quality Control in the pharmaceutical industry.
Keywords
HPLC, Method Development, Validation, Reverse Phase and Glibenclamide.
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