Bioanalytical Method Development and Validation of Doxorubicin and Lapatinib in Rat Plasma Using UHPLC-HESI-MSn

Abstract

A simple, selective, and robust method was developed to simultaneously detect doxorubicin and lapatinib in rat plasma using UHPLC-HESI-MSn based on a protein precipitation technique for extracting analytes from rat plasma. Chromatographic separation was accomplished on Waters symmetry® C18 3.5µm (4.6×75mm) column using 0.1% acetic acid in water and 0.1% acetic acid in acetonitrile as eluents followed by HESI-MSn detection. Quantifying doxorubicin and lapatinib using curcumin as an internal standard was carried out using a single reaction monitoring mode of the tandem mass spectrometer. The method is valid over the concentration range of 0.75 ng-1200 ng. This UHPLC-HESI-MSn method was fully validated based on other parameters like selectivity, matrix effect, recovery and stability. This validated method is straightforward, robust and can be clinically applicable.