Empagliflozin: Validation of Stability-Indicating LC Method and in silico Toxicity Studies.

Abstract

A new stability-indicating liquid chromatography method was developed for the quantification of empagliflozin and two synthetic impurities. The chromatographic conditions included Spherisorb® RP-18 column (150 × 4.6mm, 5μm) with a PDA detector, using acetonitrile and formic acid (pH4.0) as mobile phase in gradient elution and flow-rate of 1.2mL·min-1. The gradient increasing from 51 to 100% acetonitrile until 11.00min, followed by decreasing the solvent from 100% to the initial ratio from 11.01 to 15.00min. The method was validated according to International Council of Harmonization guidelines. The LOD and LOQ values for impurities A and B were 35 and 15ng·mL-1, respectively, (for LOD) and 115 and 35ng.mL-1, respectively (for LOQ). The method was linear in the range of 80-140, 115-1150 and 35-350ng·mL-1 for EMPA, impurities A and B, respectively, and the correlation coefficient were > 0.999 in all situations, indicating the method good linearity. The developed method showed a good recovery for empagliflozin and added impurities. The method has proven to be precise, demonstrated values less than 2.0% to empagliflozin and 5.0% to synthetic impurities, robust and selective with no interference from other products in the determination of analytes. The in silico toxicity prediction suggested that the impurities do not present any toxicity risk for the parameters evaluated.