HPTLC METHOD DEVELOPMENT AND VALIDATION FOR THE IN-VITRO INTERACTION STUDY OF MEBENDAZOLE WITH ATORVASTATIN

By the HPTLC method, a mobile phase system comprising of Ethyl acetate: Toluene: Methanol (5:4:1v/v/v) was selected, and the detection was carried out at 280 nm. The R_f values were found to be 0.65 ± 0.03 and 0.27 ± 0.03 for Mebendazole and Atorvastatin, respectively, which showed a good separation. The method was validated as per ICH guidelines. The percentage RSD values of repeatability, intraday, and interday were found to be less than 2 prove the precision of the method. The correlation coefficient value from the calibration graph was found to be 0.9949 for Mebendazole and 0.9979 for Atorvastatin. The percentage protein binding of Mebendazole and Atorvastatin was estimated by the equilibrium dialysis method. The in-vitro displacement interaction study was carried out using standard solutions of a mixture containing 1.5×10-4 M Mebendazole and Atorvastatin along with 1.5×10-4 M BSA. The percentage protein binding of individual Mebendazole and Atorvastatin after 6hr was found to be 86.14% and 91.18%, respectively. While they were evaluated in the presence of each other, the percentage protein binding was found as 84.4% for Mebendazole and 94.8% for Atorvastatin at the end of the study. The percentage protein binding of Mebendazole decreased by 1.74% and increased for Atorvastatin by 3.62%.