Bioanalytical Method Development and Validation of Bexagliflozin in Rat Plasma by LC-MS/MS Detection and its Application to A Pharmacokinetic Studies

The aim of this study is to create and verify a high throughput approach for the detection of bexagliflozin in rat plasma by LC-MS/MS detection, and to apply this approach to pharmacokinetic studies.

The Waters Alliance HPLC system was used to achieve the chromatographic separation of Bexagliflozin using D6 – Bexagliflozin as the internal standard. A Quaternary gradient pump of the e2695 series, equipped with a Q-TRAP 5500 injector with 100ng/ml, was used to inject and elute the mobile phase, which contained 0.1% Formic acid and Acetonitrile in a 50:50 v/v ratio and was pumped at a flow rate of 1 ml/min. At room temperature, absorbance at 271 nm was used as a photodiode array detector for detection.

Retention time of 4.308 minutes were used to elute the peak of bexagliflozin. Over the Bexagliflozin concentration range of 10-200 ng/ml, the standard curves were linear. The relative standard deviation of the peak areas for all measurements was determined to be less than 2.0, meeting the acceptance criteria. The mean correlation coefficient was found to be 0.999. In conclusion, the method was used for pharmacokinetic study and validation; it was shown to be stable under the analytical conditions used and to fall within acceptable bounds for linearity, accuracy, and precision.