Method Development and Validation of Simultaneous Estimation of Metolazone and Spiranolactone Using RP-HPLC Method (Research Article)

Author(s): G. Mounikha, M. Mamata*, Ch. Sampath, G. Sravanthi and P. Raj Kumar

Abstract: High performance liquid chromatography is at present one of the most sophisticated tools of the analysis. The estimation of Metolazone and Spironolactone was done by RP-HPLC. The Phosphate buffer was pH 3.0 and the mobile phase was optimized with consists of Phosphate buffer: Methanol PH 4.5(20:80 v/v). Kromosil C18 Column (250mm x 4.6mm)5μg or equivalent chemically bonded to porous silica particles was used as stationary phase. The detection was carried out using UV detector at 254 nm. The solutions were chromatographed at a constant flow rate of 0.8 ml/min. the linearity range of Metolazone and Spironolactone were found to be from 100-500 g/ml of Metolazone and 1-5g/ml of Spironolactone. Linear regression coefficient was not more than 0.999. The values of % RSD are less than 2% indicating accuracy and precision of the method. The percentage recovery varies from 98-102% of Metolazone and Spironolactone. LOD and LOQ were found to be within limit. The results obtained on the validation parameters met ICH and USP requirements .it inferred the method found to be simple, accurate, precise, and linear. The method was found to be having suitable application in routine laboratory analysis with high degree of accuracy and precision.

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