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Khusmita Patel*, Minal Rohit and Snehal Patel


The present study was attempted for development and validation of methods for the estimation of Halobetasol propionate and Miconazole nitrate in marketed formulation. This study is aimed to established a simple, precise and accurate high-performance liquid chromatography(HPLC) method for estimation of combined dosage form. The chromatographic separation was achived on column C18 Shim pack xR ODS II (25cm×4.6mm,5μm) using a mobile phase comprising of 0.2% OPA in Water: Acetonitrile in a ratio of 40:60v/v. The flow rate was 1ml/min and eluent were detected by UV detector at 291nm. Retention time were found to be 2.185 min 7.053 min of Halobetasol propionate (HALO) and Miconazole nitrate (MICO) respectively. The calibration curve was linear over the range of 8-12μg/ml for HALO and 320-480μg/ml for MICO. % Assay was found to be 98.6±0.219 and 98.35±0.065 respectively. The results of all the validation parameters were within their acceptance values. The proposed method is suitable for routine analysis of combined dosage form. Further simple, sensitive, rapid and cost effective HPTLC method was developed. The chromatographic development was carried out on aluminum plates precoated with silica gel 60 F254, using a mixture of Toluene:Methanol:TEA (8:2:0.2 v/v) was employed as mobile phase with chamber saturation time 10min. UV detection was performed at 236nm. The linear response was observed in range of 40-60ng/spot for HALO 1600- 2400ng/spot for MICO with correlation coefficient of 0.999. The percentage recovery was found to be within the limits of acceptance criteria between the range of 99.2-99.5% for HALO and 92.33-99.90% for MICO. The Rf value of analyte was found to be 0.16±0.02 for HALO and 0.32±0.02 for MICO. The proposed HPLC and HPTLC methods have adequate specificity. System precision and method precision were found to be within limits. The results of assay showed good agreement with label claim.

Keywords: Halobetasol propionate, Miconazole nitrate, High Performance Liquid Chromatographic method, High Performance Thin layer Chromatography method.

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