Fabio Bonelli
IRBM Science Park, Italy
Title: Use of dilute-and-shoot liquid chromatography high resolution mass spectrometry in preclinical research: application to a DMPK study of perhexiline in mouse plasma
Biography
Biography: Fabio Bonelli
Abstract
This work describes a simple, sensitive and rapid liquid chromatography-high resolution mass spectrometry method for quantitation of perhexiline and the simultaneous detection of perhexiline metabolites in C57b1/6 mice plasma. Only 5 µL of plasma was used for analysis. Pretreatment was limited to a 100-fold dilution (‘dilute-and-shoot’). The analyte was detected by high resolution mass spectrometry (Orbitrap Technology). Three scan events were performed over the entire chromatogram. Targeted single ion monitoring with data dependent acquisition was employed for perhexilline quantitation and confirmation, while full scan mode was used to perform untargeted detection of perhexiline phase I and phase II circulating metabolites. The calibration curve was linear (r2 = 0.990) ranging from 0.305 ng/mL (LLOQ) TO 10000 ng/mL. Matrix effect was limited to 6.1%. The method was applied to a pharmacokinetic study of perhexiline in mouse plasma and the results obtained were compared to a standard sample preparation method based on protein precipitation and liquid chromatography-tandem mass spectrometry (MRM mode) detection. The new approach provided comparable results in terms of pharmacokinetics parameters estimate with a high sensitivity, additional information on perhexiline circulating metabolites and a low consumption of biological sample. The combination of the ‘dilute-and-shoot’ approach together with HRMS targeted and untargeted detection represents a suitable alternative to classic bioanalytical approaches in preclinical research.