Development and Validation of RP-UPLC Method for Simultaneous Determination of Amiloride and Hydrochlorothiazide Using Green Assessment in Bulk and Pharmaceutical Dosage Form

The present article provides the development and validation of a RP-UPLC approach along with the GAC principles for the quantification of AML and HCTZ in bulk and in pharmaceutical dosage forms. Chromatography was performed on a 150x4.6mm Hypersil gold ODS endcapped reverse phase column with 0.1% formic acid and ethanol as mobile phase in an isocratic mode at a flow rate of 0.5 mL/min. PDA detection was done at 265 nm; AML and HCTZ were eluted with retention times of 2.98 and 5.65 min, respectively. The method was validated in accordance with ICH guidelines. The validation confirmed that the method possesses specificity, linearity, ruggedness, robustness and accuracy. Calibration plots were linear over the concentration ranges 2.5–25 μg/mL for Amiloride and 25-150 μg/mL for Hydrochlorothiazide. Limits of detection were 0.69 and 0.61 μg/mL and limits of quantification were 2.10 and 1.84 μg/mL for Amiloride and Hydrochlorothiazide, respectively. The high recovery and low coefficients of variation confirm the suitability of the method for the simultaneous determination of these two drugs in bulk and pharmaceutical dosage form. Green analytical chemistry tools are used to assess the greenness of the method and calculated using GAPI, AGREE and Eco-scale and found excellent green of >75%.