Fig. 1
Representative chromatograms of analytes: YPEIEEK (P1), GTVVVPTLDSVLYDNQEFPDPEK (P2) and internal standards for YPEIEEK (P1-IS) and GTVVVPTLDSVLYDNQEFPDPEK (P2-IS). a 30 fg µL−1 mixture of P1, P2, P1-IS and P2-IS were prepared in pooled plasma from control subjects and injected into the Agilent 1100 UHPLC coupled to the Agilent 6410 ESI-LCMS/MS system for CYP2E1 analytical method development and optimization. Stable and reproducible transition ions (m/z) were selected for MRM analysis: P1 = (precursor: 454.3; quantifier: 647.3; qualifier: 518.3); P1-IS = (precursor: 476.7; quantifier: 518.3; qualifier: 647.3); P2 = (precursor: 855.0; quantifier: 456.3; qualifier: 585.3); P2-IS = (precursor: 807.1; quantifier: 501.3; qualifier: 585.3). MRM multiple reaction monitoring; P1 peptide 1 (YPEIEEK); P1-IS internal standard for peptide 1; P2 peptide 2 (GTVVVPTLDSVLYDNQEFPDPEK); P2-IS internal standard for peptide 2. b Representative chromatogram showing good resolution of each analyte of interest from a mixture containing 30 fg µL−1 of each analyte spiked pooled plasma from control subjects. The specific retention time for each analyte was P1: 1.041 min; IS-P1: 1.811 min; P2: 2.808 min and P2-IS: 4.118 min. Total run time for each sample was 5 min