Green Assessment of Stability Indicating RP-HPLC Method for Simultaneous Estimation of Metformin, Empagliflozin, and Linagliptin in Bulk and Fixed Dosage Forms Using AGREE and GAPI Tools

Abstract

For the Simultaneous Estimation of Metformin, Empagliflozin, and Linagliptin in fixed dosage form and bulk samples, an RP-HPLC method was developed and validated in accordance with ICH guidelines. The chromatographic separation of the selected analytes was achieved using Sunfire C18 (4.6 mm x 250 mm, 5 µm) HPLC Column. The mobile phase was comprised of 0.01N Potassium dihydrogen phosphate buffer, with the pH-adjusted to 3.5 using 0.1% Ortho-phosphoric acid, and HPLC-grade acetonitrile in a ratio of 30:70 v/v. The flow rate was fixed at 1 mL/min and UV wavelength was set at 225 nm. The retention times for Metformin, Empagliflozin, and Linagliptin were 2.268 minutes, 2.822 minutes, and 3.483 minutes respectively, and the resolution between the two drugs Metformin & Empagliflozin was determined as 5.9 & for Empagliflozin, and Linagliptin it was determined as 8.2. Linearity and correlation coefficient were established for Metformin, Empagliflozin, and Linagliptin in the concentration range of 50-300 µg/mL with r2 =0.9995, 1.25-7.5 µg/mL with r2=0.9997, and 0.25-1.5 µg/mL with r2=0.993 respectively. Metformin, Empagliflozin, and Linagliptin exhibited the percentage recoveries of 100.02 %, 99.98 %, and 99.82 % correspondingly. The detection limit values obtained were 3.25 µg/mL, 0.08 µg/mL, and 0.01 µg/mL & the quantification limit values obtained were 9.86 µg/mL, 0.25 µg/mL, 0.04 µg/mL for Metformin, Empagliflozin and Linagliptin correspondingly. The developed HPLC approach was also applied for force degradation studies and the green assessment was also determined using two tools AGREE & GAPI. The optimized method was found to be both cost-effective, environment-safe and user-friendly due to its green profile, reduced retention times & stability, rendering it appropriate for routine quality control testing in Pharmaceutical industries.