Stability indicating Method Development and Validation for simultaneous estimation of Hydrochlorothiazide, Amlodipine and Olmesartan in tablet dosage form by using RP-HPLC

The present work was undertaken with the aim to develop and validate a rapid and consistent RP-HPLC method in which the peaks will be appear with short period of time as per ICH Guidelines. The proposed method was simple, fast, accurate and precise method for the Quantification of drug in the dosage form, bulk drug as well as for routine analysis in Quality control.

RP-HPLC method was developed and validated for simultaneous estimation of Hydrochlorothiazide, Amlodipine and Olmesartan in bulk drug and in combined dosage forms. The HPLC separation was achieved on a Symmetry C18 (4.6 X 150mm, 5□m, Make: XTerra) or equivalent in an Isocratic Mode. The mobile phase was composed of TEA Buffer (40%) whose pH was adjusted to 3.5 by using Ortho Phosphoric Acid and Acetonitrile (60%) [HPLC Grade] The flow rate was monitored at 0.8ml per min. The wavelength was selected for the detection was 230 nm. The run time was 9 min.

The retention time found for the drugs Hydrochlorothiazide, Amlodipine and Olmesartan were 3.034 min., 4.062 min. and 5.165 min. respectively. The % recovery was found to be 99.3% -101.7% for the drug Hydrochlorothiazide. The % recovery was found to be 98.3% -99.3% for the drug Amlodipine. The % recovery was found to be 98.3% -100.7% for the drug Olmesartan. The linearity was established in the range of 25 to 62.5ppm for the drug Hydrochlorothiazide and 10 to 25ppm for the drug Amlodipine and 10 to 100 ppm for the drug Olmesartan. The LOD for the drugs Hydrochlorothiazide, Amlodipine and Olmesartan were found to be 0.009μg/ml, 0.06μg/ml and 0.06μg/ml respectively. The LOQ for the drugs Hydrochlorothiazide, Amlodipine and Olmesartan were found to be 0.03μg/ml, 0.2μg/ml and 0.2μg/ml respectively.

The proposed method was adequate sensitive, reproducible, and specific for the determination of Hydrochlorothiazide, Amlodipine and Olmesartan in bulk as well as in Tablet dosage form. The validation of method was carried out utilizing ICH-guidelines. The described RP-HPLC method was successfully employed for the analysis of pharmaceutical formulations containing combined dosage form. Overall the proposed method was found to be suitable and accurate for the Quantitative determination of the drug in Tablet dosage form. The method was simple, precise, accurate and sensitive and applicable for the simultaneous determination of Hydrochlorothiazide, Amlodipine and Olmesartan in bulk drug and in combined dosage forms.