Abstract
UHPLC-MS/MS DETERMINATION OF GLP-1 ANALOGUE, LIRAGLUTIDE A BIOACTIVE PEPTIDE IN HUMAN PLASMA

Mubarak Hasan Shah*, Padmanabha Reddy Yeragam Reddy, Madhira Bhawanishankar, Mokkaisamy Jegadeesh Raja and Manoj Pillai

ABSTRACT

Liraglutide is a potent GLP-1 agonist that exhibits hypoglycemic effect for type 2 diabetes mellitus. In the present work, a quantitative method to analyze liraglutide in human plasma was developed and validated by UHPLC-MS/MS following international guidelines. The method uses an internal standard, leuprolide for quantification. Sample processing includes solid phase extraction to remove the most abundant plasma proteins and the recovery of liraglutide peptide was found be consistent across the tested concentrations. Quantification was achieved by positive electrospray ionization containing MRM transitions of m/z 1251.3→1245.4 for liraglutide and 605.6→249.1 for IS using API 6500 QTRAP mass spectrometer. The lower limit of quantification was 0.5 pg/mL and the assay exhibited a linear range of 0.5 to 100 pg/mL. The intra- and inter-day precisions were within 11.62% CV, while the accuracy was within ±7.6% of nominal values. The calibration curves showed R2 higher than 0.99 across different batches. The stability of the analyte was found to be within acceptable limits where the samples would experience in a real time analysis.

Keywords: Liraglutide, Ultra high performance liquid chromatography tandem mass spectrometry, human plasma.


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