Development and Validation of UHPLC‐MS/MS Method for the Determination of SAX and 5‐OH‐SAX in Human Plasma and Its Application to a Pharmacokinetic Study

A highly sensitive and reliable ultrahigh performance liquid chromatography-tandem mass spectrometry method (UHPLC-MS/MS) has been developed for the quantification of saxagliptin (SAX) and its metabolite, 5-hydroxy saxagliptin (5-OH-SAX), in human plasma. Utilizing an ACE Excel 3 Super C18 column with electrospray ionization multiple reaction monitoring, this method achieves excellent linearity across the concentration ranges of 0.1-50 ng/mL for SAX and 0.2-100 ng/mL for 5-OH-SAX. The mean extraction recoveries are reported at 99.87% for SAX and 97.68% for 5-OH-SAX, with matrix effects maintained at 0.85 and 0.86, respectively. Both intraday and interday relative standard deviations are maintained below 15%. Furthermore, the developed method confirms appropriate short-term stability, satisfying the requirements for pharmaceutical analysis without residual effects. Significantly, this validated method applies to pharmacokinetic studies, providing crucial insights into the drug and metabolite behaviors during varying ingestion states, thus enhancing therapeutic efficacy and patient compliance through informed SAX administration.