A new RP-HPLC method for separation and determination of process related impurities in Pioglitazone Hydrochloride API

摘要

The objective of the study was to develop a validated, specific and stability-indicating gradient reverse phase liquid chromatographic method for determination of Pioglitazone Hydrochloride along with its impurities in bulk samples. Drug substance was subjected to stress conditions of hydrolysis (acid and base), oxidation, photolytic, humidity and thermal degradation as per International Conference on Harmonization (ICH) to show the stability-indicating power of the method. Significant degradation was observed with alkali and hydrogen peroxide. The impurities were characterized using spectral techniques like IR, ~1H NMR and MS. Successful separation of impurities was achieved on C18 ODS (150×4.6 mm) 3.5 μ column using mobile phase consisting of Solvent A: Ammonium acetate buffer and Acetonitrile in the ratio (57:43 v/v) for 0-7 min and Solvent B: Ammonium acetate buffer and Acetonitrile in the ratio (20:80 v/v) at a flow rate of 1.0 ml/min from 7-20 min followed by Solvent A from 20-21 min. The retention times of impurity A, impurity B, impurity C and Pioglitazone were 3.44, 10.65, 17.95 and 8.32 min respectively. The detection wavelength was set at 254 nm with column temperature at 45 ℃. Finally developed method was validated as per ICH guidelines for specificity, linearity, precision and accuracy.