Method Development and Validation for the Quantification of Abametapir in Biological Matrices by LC-ESI-MS/MS

Abstract

Aim A precise, linear and specific liquid chromatography-tandem mass spectrometry procedure was executed and subjected for validation for the quantification of plasma. Materials and Methods The chromatography elution was carried out using a C18-Discovery column of 15 cm in length and 2.1 mm in internal diameter, packed with stationary phase particles of 5 µm size, at a flow rate of 0.80 mL/min. An isocratic elution process was conducted with a mobile phase solution consisting of methanol and 0.10% V/V HCOOH in a ratio of 90:10. The separation of Abametapir and Metformin was achieved by a liquid-liquid extraction process using ethyl acetate as the solvent. Results A triple quadrupole mass spectrometer was used for the measurement of ions. Electrospray ionization is a technology that ionizes positively, and it was used in Multiple Reaction Monitoring (MRM) with parent/product ionic transitions m/z 185.1 →106.06 for Abametapir and 130.1 →60.20 for the Metformin internal standard. A calibration graph was constructed with values ranging from 2.05 to 82.00 ng/mL, resulting in the equation y=0.0149x+0.00221, with a r2 value above 0.99. The recovery values of Abametapir exceeded 94.27%, with its accuracy assessed in terms of relative error ranging from -4.04% to 6.52%. Conclusion The accuracy, recovery and sensitivity values of Abametapir in the plasma sample, as shown by the established approach, highlight its significance in pharmacokinetic and bioequivalence research.

Keywords: LC-MS/MS; Abametapir; Metalloproteinase Inhibitor; Validation; Accuracy; Linearity