Development and validation of a UPLC-HR-SIM method to quantify semaglutide for its pharmacokinetic study

To develop a highly sensitive and interference-resistant ultra performance liquid chromatography (UPLC) coupled with high-resolution selected ion monitoring (HR-SIM) mass spectrometry method for the quantification of semaglutide in biological samples, and to apply it for pharmacokinetic analysis. The UPLC-HR-SIM method was utilized to quantify semaglutide in beagle plasma, requiring minimal sample pretreatment. The method demonstrated a lower limit of quantification (LLOQ) of 5.0 ​ng/mL for semaglutide in beagle plasma. Following intravenous (iv) administration at a dose of 0.030 ​mg/kg, the plasma concentration of semaglutide in beagles exhibited a multi-exponential decay pattern, with an average elimination half-life (t 1/2 ) of 44.90 ​± ​11.45 ​h. The UPLC-HR-SIM method has proven to be a highly sensitive and robust approach for the quantification of peptide-based drugs. This method will enhance the understanding of the pharmacokinetics and pharmacodynamics of semaglutide and facilitate further research into the pharmacokinetics of other peptide therapeutics. • A highly sensitive and universal method for the quantitation of peptide drugs. • This method has been fully validated for semaglutide quantifying. • Pharmacokinetic parameters of somaglutide in the plasma of beagles.