Journal of Bioequivalence & Bioavailability

Abstract

Novel LC-MS/MS Method for Quantification of Semaglutide in Human Plasma and its Application in Pharmacokinetic Studies

Arjun Arumugam, Geetha Lakshmi, Nageswara Rao Thalapaneni* and Srinivas Gopineedu

The objective of the study was to develop a bioanalytical method for the determination of semaglutide in human K₃ EDTA plasma. Separation was achieved by reverse phase chromatography, and quantification was done using the LC-MS/MS method. The chromatographic separation was achieved with an phenomenex aeris wide pore XB-C8 (100 × 4.6 mm) 3.6 μm column thermostated at 40°C with a mobile phase containing gradient program with Pump A: (Acetonitrile: Methanol (50:50) v/v) and Pump B: (0.2% Formic acid in Milli Q Water (v/v)). The flow rate was maintained at 0.800 ml/min, and the injection volume was found to be 5 μl. The detection was done by using high-performance liquid chromatography coupled with tandem mass spectrometry. The retention time was found to be 5.40 ± 0.80 min for the analyte and 5.40 ± 0.80 min for the internal standard. The calibration curves showed good linearity (r2>0.99) over concentration range of 2-120 ng/mL. The precision (2.42% to 7.78%) and accuracy (95.67% to 104.93%) fell within the range, all meeting acceptance criteria. The recovery of analyte 67.51% was well within the acceptance limits. The established analytical method was found to be selective, sensitive, precise, accurate, reproducible and validated according to the International Conference on Harmonization guidelines.

Published Date: 2024-12-16; Received Date: 2024-11-16