Abstract :
Introduction: Analytical method development and validation are crucial for ensuring drug quality and efficacy. As combination therapy is common in type 2 diabetes, a reliable method is needed for the simultaneous estimation of Metformin, Sitagliptin, and Imeglimin in pharmaceutical formulations. This study aims to develop and validate a simple, precise, and robust RP-HPLC-UV method for the simultaneous estimation of Metformin, Sitagliptin and Imeglimin in single combined dosage forms.
Materials & Methods: Metformin, Sitagliptin, and Imeglimin were analyzed using a Shimadzu RP-HPLC system with a C18 column and UV detection at 260 nm. Stock and working solutions were prepared in aqueous media, and calibration curves were constructed over relevant concentration ranges. The optimized mobile phase (Methanol: Cyanomethane: 15 mM Ammonium Formate, 10:40:50 v/v, pH 6.5) under isocratic flow (0.9 mL/min) provided well-resolved peaks with retention times of 3.7, 4.7, and 8.3 min. The method was validated as per ICH guidelines for specificity, linearity, accuracy, precision, LOD, LOQ, robustness, and ruggedness.
Results: The developed RP-HPLC-UV method enabled efficient and simultaneous estimation of Sitagliptin, Metformin, and Imeglimin with well-resolved peaks at retention times. The method exhibited high specificity, with no interference from diluents or other components. The LOD and LOQ were 85/265 ng/mL for Sitagliptin, 25/80 ng/mL for Metformin, and 55/175 ng/mL for Imeglimin. Accuracy, assessed via recovery studies, ranged from 98.6–99.9%, and precision studies showed low %RSD, confirming repeatability and reproducibility. System suitability parameters, including theoretical plates, tailing factor, and resolution, were within acceptable limits, indicating the method’s reliability for routine analysis of these antidiabetic drugs.
Conclusions: A validated RP-HPLC-UV method was developed for simultaneous estimation of Sitagliptin, Metformin, and Imeglimin. The method showed high specificity, accuracy, and precision, suitable for routine pharmaceutical analysis.

Keyword :Analytical method, Imeglimin, Metformin, RP-HPLC, Sitagliptin and Validation.