Analytical Method Development and Validation of Simultaneous Estimation of Ivermectin and Albendazole Byusing RP-HPLC

A new method was established for simultaneous estimation of Ivermectin and Albendazole by RP-HPLC method. The chromatographic conditions were successfully developed for the separation of Ivermectin and Albendazole by using ACE C18 column (4.6×150 mm) 5µ, flow rate was 1.2 ml/min, mobile phase ratio was (70:30 v/v) methanol: Phosphate buffer pH 3 (pH was adjusted with ortho phosphoricacid), detection wavelength was 240 nm. The instrument used was WATERS HPLC Auto Sampler, Separation module 2690, photodiode array detector 996, Empower-software version-2. The retention times were found to be 2.344 mins and 3.284 mins. The % purity of Ivermectin and Albendazole was found to be 101.27% and 99.97% respectively. The system suitability parameters for Ivermectin and Albendazole such as theoretical plates and tailing factor were found to be 4668, 1.3 and 6089 and 1.2, the resolution was found to be 6.0. The analytical method was validated according to ICH guidelines (ICH, Q2 (R1)). The linearity study n Ivermectin and Albendazole was found in concentration range of 50 µg-250 µg and 5µg-50µg and correlation coefficient (r2) was found to be 0.999 and 0.999, % recovery was found to be 99.56% and 99.48%, % RSD for repeatability was0.2 and 0.2, % RSD for intermediate precision was 0.2 and 0.1respectively. The precision study was precise, robust, and repeatable. LOD value was 3.17 and 5.68, and LOQ value was 0.0172 and 0.2125 respectively. Hence the suggested RP-HPLC method can be used for routine analysis of Ivermectin and Albendazole in API and Pharmaceutical dosage form.