Liquid chromatography-mass spectrometry for quantification of the anti-chemotherapeutic antibody in rat plasma / 国际药学研究杂志

ABSTRACT

Objective To establish a sensitive, accurate and reliable LC-MS method to quantify the monoclonal antibody in rat blood and preclinical pharmacokinetics study. Methods The method of selective reaction monitoring (SRM) was established by using on-the-fly orthogonal array optimization (OAO) with anti-chemotherapeutic antibody-06(ACA-06)as a model drug, and the pharmacokinetics of ACA-06 in rats was studied. The optimal signature peptide selection and mass spectrometry conditions were optimized by single LC-MS run. Two highly specific and sensitive peptides, FSGVPDR and VNSAAFPAPIEK, were identified as the signature peptides of quantification based on the OAO optimization result, which were located in the ACA-06 light chain and heavy chain complementary determining region (CDR) respectively. Results The method had good linearity in the range of 12.9~32 250 ng/ml (r20.9819-0.9946). The lower limit of quantification was 12.9 ng/ml. The intra-day precision (RSD) was less than 11.5%, the inter-day RSD was less than 6%, and the accuracy was between -4.0% and 4.3%. Conclusion In this study, LC/SRM-MS method is established for the determination of ACA-06 in rat plasma. The method is specific, accurate and sensitive and enables an accurate pharmacokinetics investigation of ACA-06 in rat plasma.