title: High-Throughput Chiral LC-MS/MS Method Using Overlapping Injection Mode for the Determination of Pantoprazole Enantiomers in Human Plasma with Application to Pharmacokinetic Study.
abstract: A sensitive and high-throughput chiral liquid chromatography-tandem mass spectrometry method was developed and validated for the quantification of R-pantoprazole and S-pantoprazole in human plasma. Sample extraction was carried out by using ethyl acetate liquid-liquid extraction in 96-well plate format. The separation of pantoprazole enantiomers was performed on a CHIRALCEL OJ-RH column and an overlapping injection mode was used to achieve a run time of 5.0 min/sample. The mobile phase consisted of 1) 10 mM ammonium acetate in methanol: acetonitrile (1:1, v/v) and 2) 20 mM ammonium acetate in water. Isocratic elution was used with flow rate at 500 μL/min. The enantiomers were quantified on a triple-quadrupole mass spectrometer under multiple reaction monitoring (MRM) mode with m/z 382.1/230.0 for pantoprazole and m/z 388.4/230.1 for pantoprazole-d7. Linearity from 20.0 to 5000 ng/mL was established for each enantiomer (r(2)  > 0.99). Extraction recovery ranged from 91.7% to 96.4% for R-pantoprazole and from 92.5% to 96.5% for S-pantoprazole and the IS-normalized matrix factor was 0.98 to 1.07 for R-pantoprazole and S-pantoprazole, respectively. The method was demonstrated with acceptable accuracy, precision, selectivity, and stability and the method was applied to support a pharmacokinetic study of a phase I clinical trial of racemic pantoprazole in healthy Chinese subjects. Chirality 28:569-575, 2016. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, People's Republic of China.|Bioanalytical Services, WuXi AppTec Co., Ltd., Waigaoqiao Free Trade Zone, Shanghai, People's Republic of China.
authors.forename: Shengni
authors.initials: S
authors.lastname: Li
authors.affiliation: Bioanalytical Services, WuXi AppTec Co., Ltd., Waigaoqiao Free Trade Zone, Shanghai, People's Republic of China.
authors.forename: Huafang
authors.initials: H
authors.lastname: Jiang
authors.affiliation: Bioanalytical Services, WuXi AppTec Co., Ltd., Waigaoqiao Free Trade Zone, Shanghai, People's Republic of China.
authors.forename: Yiya
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, People's Republic of China.
authors.forename: Yinli
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Bioanalytical Services, WuXi AppTec Co., Ltd., Waigaoqiao Free Trade Zone, Shanghai, People's Republic of China.
authors.forename: Xiaohang
authors.initials: X
authors.lastname: Shen
authors.affiliation: Bioanalytical Services, WuXi AppTec Co., Ltd., Waigaoqiao Free Trade Zone, Shanghai, People's Republic of China.
authors.forename: Wenzhong
authors.initials: W
authors.lastname: Liang
authors.affiliation: Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, People's Republic of China.
authors.forename: Zhanying
authors.initials: Z
authors.lastname: Hong
mesh_terms: D053799:2-Pyridinylmethylsulfinylbenzimidazoles / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics*; D000328:Adult; D002853:Chromatography, Liquid / Q000379:methods*; D004355:Drug Stability; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D059625:Liquid-Liquid Extraction; D000077402:Pantoprazole; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D053799:2-Pyridinylmethylsulfinylbenzimidazoles; D000077402:Pantoprazole
keywords: LC-MS/MS; chiral separation; human plasma; overlapping injection; pantoprazole enantiomers