Quantification Of Semaglutide In Human Plasma Using Xevo™ TQ-XS Mass Spectrometer: A Highly Sensitive And Reliable Analytical Method

By Tirumal Datar, Sushil Gawande, Dr. Padmakar Wagh, and Amy Bartlett, Waters Corporation

In this application note, we detail the development and validation of a highly sensitive and robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the precise quantification of Semaglutide in human plasma. The analytical method utilizes the Xevo TQ-XS tandem quadrupole mass spectrometer in conjunction with the ACQUITY™ Premier UPLC™ system, which offers enhanced performance in both detection and separation. Chromatographic separation was achieved using the ACQUITY Premier Peptide BEH™ C18 Column, which was carefully selected for its efficiency in handling peptide-based analytes. A key innovation in this method is the incorporation of difluoroacetic acid (DFA) as an ion-pairing reagent, which significantly improves peak shape and boosts ionization efficiency for charged analytes, resulting in sharper, more defined peaks and heightened sensitivity. The method underwent rigorous validation to ensure its reliability and applicability in clinical and research settings.

Find out how it demonstrated excellent selectivity and reproducibility, with a linear response observed across a clinically relevant concentration range of 0.1 ng/mL to 20 ng/mL.