Simultaneous, Development and Validation Measurement of Empagliflozin and Linagliptin for Glyxambi® Tablets using TLC and RP-HPLC

This aims of study to develop and validate two chromatographic methods that provide a fast, sensitive, and reliable stability-indicating assay for the simultaneous determination of Empagliflozin and Linagliptin in their pure forms, as well as Glyxambi® tablets. The first technique employed was thin-layer chromatography (TLC) utilizing a developing solution composed of ethyl acetate, methanol, and hexane in a ratio of 90:5:5 (v/v/v). The separation of Empagliflozin and Linagliptin was achieved on silica gel plates, with a scanning wavelength at 248 nm. Retention factor (RF) values for Linagliptin was 0.16 and Empagliflozin was 0.51. Mean percentage recoveries were 101.32% (SD 0.942) for Linagliptin and 99.84% (SD 1.426) for Empagliflozin, the linearity across concentration ranges of 5-50µg/band and 10-150µg/band, respectively. The second technique, known as RP-HPLC, separated the samples using a GL Sciences reverse-phase C18 column (250mm x 4.6mm x 5µm). Water, acetonitrile, and 0.05 M phosphate buffer with a pH of 3.8 (45:50:5, v/v/v) made up the mobile phase, which was used for 10min. We carried out UV detection at 248nm. The chromatogram showed the presence of Linagliptin and Empagliflozin, with retention periods of Rt = 2.242min and 4.065min, respectively. Mean percentage recoveries for Linagliptin were 100.81% (SD 1.631) and for Empagliflozin was 101.44% (SD 1.381), A satisfactory resolution was attained, and linearity was established within the range of 0.5-10 and 2-35 µg/ml, respectively. We successfully determined Linagliptin and Empagliflozin, whether in bulk powder or combination dose form (Glyxambi® tablets), quantitatively for the pharmaceutical market using the TLC and RP-HPLC techniques.