DEVELOPMENT AND VALIDATION OF ANALYTICAL METHOD FOR SIMULTANEOUS ESTIMATION OF TENELIGLIPTIN HYDROBROMIDE HYDRATE, PIOGLITAZONE HYDROCHLORIDE, AND METFORMIN HYDROCHLORIDE IN PHARMACEUTICAL DOSAGE FORM

Abstract

A simple, precise, and accurate RP-HPLC method was developed and validated for the simultaneous estimation of TENE, PIO, and MET in pharmaceutical formulations as per ICH guidelines. The analytical wavelength utilized was 254.8 nm with linearity ranges of 1-6 μg/mL (TENE), 0.75-4.5 μg/mL (PIO), and 25-150 μg/mL (MET). Good linearity was demonstrated by the regression equations and correlation coefficients (r2 > 0.9985). Percentage recovery studies at 80%, 100%, and 120% levels showed mean recoveries within 99.5-102.3% (TENE), 99.4-102.2% (PIO), and 99.2-100.4% (MET) with relative standard deviation (RSD) < 1.4%, indicating accuracy. Precision was evaluated by intraday, interday, and repeatability studies, revealing RSD < 1.7% (TENE), < 0.9% (PIO), and < 1.1% (MET), suggesting good precision. Limits of detection were 0.242 μg/mL (TENE), 0.150 μg/mL (PIO), and 0.143 μg/mL (MET), while limits of quantitation were 0.732, 0.454, and0.434 μg/mL, respectively. The robustness of the RP-HPLC method was evaluated using a fractional factorial design by varying method parameters such as flow rate, mobile phase composition, and detection wavelength. The results demonstrated the method's resilience to minor parameter changes, indicating its robustness.