Simultaneous quantification of EVT201, a novel partial positive allosteric GABAA receptor modulator, and its two metabolites in human plasma by UHPLC/MS/MS

Publication date: Available online 1 June 2018Source: Journal of Pharmaceutical and Biomedical AnalysisAuthor(s): Xianhua Zhang, Congya Zhou, Yuanyuan Zhang, Shuangpeng Zhu, Xiaofeng Cai, Chunmiao Pan, Chuanzhen Wang, Suodi Zhai, Li YangAbstractA sensitive, accurate and rapid UHPLC-MS/MS method was developed and validated for the simultaneous determination of EVT201 and its two metabolites, Ro46-1927 and Ro18-5528, in human plasma. D6-EVT201 was used as the internal standard (IS). Plasma samples were extracted using ethyl acetate after being alkalized with saturated sodium carbonate solution. Chromatographic separation was carried out on an Acquity BEH C18 column (2.1 × 50 mm, 1.7 μm) with a gradient mobile phase at a flow-rate of 0.50 mL/min. The analytical run time was 5.5 min. For mass spectrometric detection, multiple reaction monitoring (MRM) was used. The MRM ion transitions were m/z 373.2 → 58.0 for EVT201, m/z 359.2 → 316.1 for Ro46-1927, m/z 291.2 → 274.1 for Ro18-5528 and m/z 379.3 → 64.0 for the IS. The linear range was 0.2–200 ng/mL for each analyte, with a correlation coefficient (r) over 0.9900. The intra-/inter- precision was within 7.9% and 4.5% for EVT201, 13.2% and 6.3% for Ro46-1927, 3.7% and 4.1% for Ro18-5528. For the accuracy, the relative bias of intra-/inter- run was no more than 6.4% and 4.6% for EVT201, 9.4% and 7.9% for Ro46-1927, −6.9% and −7.5% for Ro18-5528. The validated method was successfully appli...
Source: Journal of Pharmaceutical and Biomedical Analysis - Category: Drugs & Pharmacology Source Type: research