title: Development and validation of a liquid chromatography/tandem mass spectrometry assay for the simultaneous determination of dabigatran etexilate, intermediate metabolite and dabigatran in 50μL rat plasma and its application to pharmacokinetic study.
abstract: A simple, rapid and specific high performance liquid chromatography-electrospray ionization tandem mass spectrometry method was developed for simultaneous determination of dabigatran etexilate (BIBR 1048 MS), the intermediate metabolite (BIBR 1087 SE) and dabigatran (BIBR 953 ZW). In this method, a stacked protein precipitation with methanol was performed in Sirocco 96-well filtration plates to extract analytes using only 50μL plasma. The analysis was performed on an Ultimate TM XB-C18 (4.6×50mm, 5μm) column using gradient elution with a mobile phase composed of methanol containing 0.01% formic acid and pure water at a flow rate of 0.3mL/min. The gradient was set to 90% methanol containing 0.01% formic acid for the first 1.0min, after which it dropped to 10%, and then was kept at 10% for the next 5min followed by an additional 1.0min at the initial composition of 90% methanol containing 0.01% formic acid for equilibration. Detection was performed on a triple-quadrupole mass spectrometer electrospray ionization interface in positive ion mode. Linear calibration curves were obtained over the concentration ranges of 1-500ng/mL for all analytes. The validated LC-MS/MS method for its selectivity, sensitivity, linearity, precision, accuracy, recovery, matrix effect and stability had been successfully applied to a pharmacokinetic study of analytes in rat plasma following a single oral administration of 15mg/kg dabigatran etexilate.
authors.affiliation: Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, No. 325 Guohe Road, Shanghai 200433, PR China; Shanghai Key Laboratory for Pharmaceutical Metabolite Research, No. 325 Guohe Road, Shanghai 200433, PR China; Xuhui Central Hospital of Shanghai, No. 966 Huaihai Road, Shanghai 200031, PR China.
authors.forename: Jinyin
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, No. 325 Guohe Road, Shanghai 200433, PR China; Shanghai Key Laboratory for Pharmaceutical Metabolite Research, No. 325 Guohe Road, Shanghai 200433, PR China; Heilongjiang Institute for Food and Drug Control, Haerbin 150001, PR China.
authors.forename: Jing
authors.initials: J
authors.lastname: Fang
authors.affiliation: College of Science, China Agricultural University, Beijing 100083, PR China.
authors.forename: Fanyi
authors.initials: F
authors.lastname: Zhong
authors.affiliation: Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, No. 325 Guohe Road, Shanghai 200433, PR China; Shanghai Key Laboratory for Pharmaceutical Metabolite Research, No. 325 Guohe Road, Shanghai 200433, PR China; School of Pharmacy, Fujian University of Traditional Chinese Medicine, Fujian 350108, PR China.
authors.forename: Wenxue
authors.initials: W
authors.lastname: Li
authors.affiliation: Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, No. 325 Guohe Road, Shanghai 200433, PR China; Shanghai Key Laboratory for Pharmaceutical Metabolite Research, No. 325 Guohe Road, Shanghai 200433, PR China.
authors.forename: Yuanjun
authors.initials: Y
authors.lastname: Tang
authors.affiliation: Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, No. 325 Guohe Road, Shanghai 200433, PR China.
authors.forename: Yungen
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Xuhui Central Hospital of Shanghai, No. 966 Huaihai Road, Shanghai 200031, PR China. Electronic address: shilongmao1965@126.com.
authors.forename: Shilong
authors.initials: S
authors.lastname: Mao
authors.affiliation: Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, No. 325 Guohe Road, Shanghai 200433, PR China; Shanghai Key Laboratory for Pharmaceutical Metabolite Research, No. 325 Guohe Road, Shanghai 200433, PR China. Electronic address: guorfan@outlook.com.
authors.forename: Guorong
authors.initials: G
authors.lastname: Fan
keywords: Dabigatran; LC–MS/MS; Method validation; Pharmacokinetic study; Sirocco 96-well filtration plates