Bioanalytical Method Development Of Tirzepatide In Rat Plasma Using Lixisenatide As Internal Standard Using Liquid Chromatography Coupled With Tandem Mass Spectroscopy And Application To Pharmacokinetic Studies

Abstract

A rapid, sensitive and reproducible LC-MS/MS was developed and validated for the quantification of Tirzepatide in Rat plasma using Lixisenatide as internal standard. The reconstituted samples were chromatographed on a Phenyl, 150mm x 4.6mm, 3.5µm column using a Methanol: 0.1% Formic acid 50:50 v/v as the mobile phase at a flow rate of 1.0 mL/min. Good response was found in positive ion mode using MRM technique. The total chromatographic run time was set at 5 min. The calibration curve was found to be linear over the concentration range of 2.50–50.00 ng/mL for Tirzepatide with a coefficient of correlation of >0.99. No significant matrix effect was observed in all the six batches of rat plasma for the analyte at LQC and HQC concentrations. Accuracy, precision, recovery, matrix effect and stability results were found to be within the suitable limits. Simple and efficient method was developed and utilized in pharmacokinetic studies to see the investigated analyte in body fluids. The application denotes all the parameters of system suitability, specificity, linearity and accuracy are in good agreement with USFDA guidelines and applied effectively for the investigation of pharmacokinetic studies in rat