Abstract
HPLC method was developed for the simultaneous estimation of Losartan Potassium, Ramipril and Hydrochlorothiazide in bulk as well as in pharmaceutical dosage form using Symmetry C18 column (4.6 x 150mm, 5mm, Make: Hypersil) in isocratic mode. SUMMARY: The mobile phase was consisting of Potassium dihydrogen phosphate (KH2PO4) and Acetonitrile [HPLC Grade] in the ratio of 68:32(%v/v). The detection was carried out at 210 nm. The % mean recoveries of Losartan Potassium, Ramipril and Hydrochlorothiazide were found to be 98.0 to 100.8%, 98.9-100.7%and 98.3 to 101.2% respectively. The method was linear over the concentration range for Losartan 50.0 ppm-110.0 ppm, Ramipril 1.255ppm-2.75ppm & Hydrochlorothiazide 12.5ppm-27.5 ppm. The Limit of Detection and Limit of Quantification of the method were calculated basing on standard deviation of the response and the slope (s) of the calibration curve at approximate level. The Limit of Detection for Losartan, Ramipril and Hydrochlorothiazide were found to be 0.07, 0.078 &.01μg/mL respectively and Limit of Quantification for Losartan, Ramipril and Hydrochlorothiazide were found to be 0.24, 0.27 and 0.05μg/mL respectively. The bulk active pharmaceutical ingredient was subjected to thermal, photolytic, hydrolytic (acidic and basic) and oxidative stress conditions and stressed samples were analyzed by the proposed method. The resultant stressed samples were analyzed by the proposed method and was established to provide high resolution among the degradation products and the analyte. All the peaks of degraded product were resolved from the active pharmaceutical ingredient with significantly different retention time and the peak purity of analyte peaks in the stressed samples was confirmed by photodiode array detector. CONCLUSION: The validation of method was carried out utilizing ICH-guidelines. The described HPLC method was successfully employed for the analysis of pharmaceutical formulations containing combined dosage form. The method was effectively separated the drug from its degradation product and it was employed as a stability- indicating one.
