Simultaneous Characterization and Determination of Warfarin and Its Hydroxylation Metabolites in Rat Plasma by Chiral Liquid Chromatography-Tandem Mass Spectrometry
Warfarin is extensively used for venous thromboembolism and other coagulopathies. In clinical settings, warfarin is administered as a mixture of S- and R-warfarin, and both enantiomers are metabolized by multiple cytochrome P450 enzymes into many hydroxylation metabolites. Due to the high degree of...
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MDPI AG
2022-05-01
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| Series: | Pharmaceutics |
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| Online Access: | https://www.mdpi.com/1999-4923/14/6/1141 |
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| author | Shasha Jin Zhihong Li Qing Yang Boyu Fang Xiaoqiang Xiang Chao Peng Weimin Cai |
| author_facet | Shasha Jin Zhihong Li Qing Yang Boyu Fang Xiaoqiang Xiang Chao Peng Weimin Cai |
| author_sort | Shasha Jin |
| collection | DOAJ |
| description | Warfarin is extensively used for venous thromboembolism and other coagulopathies. In clinical settings, warfarin is administered as a mixture of S- and R-warfarin, and both enantiomers are metabolized by multiple cytochrome P450 enzymes into many hydroxylation metabolites. Due to the high degree of structural similarity of hydroxylation metabolites, their profile possesses significant challenges. The previous methods generally suffer from lacking baseline resolution and/or involving complex analysis processes. To overcome this limitation, a sensitive and specific chiral liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed to simultaneously identify warfarin and hydroxywarfarins enantiomers. Chromatographic separation was achieved on a HYPERSIL CHIRAL-OT column. The mass spectrometric detection was carried out in negative ion MRM mode with electrospray ionization source. The optimized method exhibited satisfactory within-run and between-run accuracy and precision with lower limit of quantification (LLOQ) of 10.0 ng/mL and 1.0 ng/mL for warfarin and 7-, 10(R)-OH-warfarin enantiomers, respectively. Linear responses of warfarin enantiomers and 7-, and 10(R)-OH-warfarin enantiomers in rat plasma were observed over the range of 10.0–8000 ng/mL, and 1.00–800 ng/mL, respectively. The analytes were shown to be stable in various experimental conditions in rat plasma. Protein precipitation was used in sample preparation without a matrix effect. This method was successfully applied to pharmacokinetic study for quantitating the concentrations of S/R-warfarin, S/R-7-OH-warfarin, and S/R-10(R)-OH-warfarin and relatively quantitating 3′-, 4-, 6-, and 8-OH warfarin enantiomers in rat plasma. |
| first_indexed | 2024-03-09T22:46:22Z |
| format | Article |
| id | doaj.art-538b014b43444bb18b1472b4fba1c059 |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-09T22:46:22Z |
| publishDate | 2022-05-01 |
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| series | Pharmaceutics |
| spelling | doaj.art-538b014b43444bb18b1472b4fba1c0592023-11-23T18:28:53ZengMDPI AGPharmaceutics1999-49232022-05-01146114110.3390/pharmaceutics14061141Simultaneous Characterization and Determination of Warfarin and Its Hydroxylation Metabolites in Rat Plasma by Chiral Liquid Chromatography-Tandem Mass SpectrometryShasha Jin0Zhihong Li1Qing Yang2Boyu Fang3Xiaoqiang Xiang4Chao Peng5Weimin Cai6Department of Clinical Pharmacy, School of Pharmacy, Fudan University, Shanghai 201203, ChinaNational Facility for Protein Science in Shanghai, Zhangjiang Lab, Shanghai Advanced Research Institute, Chinese Academy of Sciences, Shanghai 201210, ChinaDepartment of Clinical Pharmacy, School of Pharmacy, Fudan University, Shanghai 201203, ChinaDepartment of Clinical Pharmacy, School of Pharmacy, Fudan University, Shanghai 201203, ChinaDepartment of Clinical Pharmacy, School of Pharmacy, Fudan University, Shanghai 201203, ChinaNational Facility for Protein Science in Shanghai, Zhangjiang Lab, Shanghai Advanced Research Institute, Chinese Academy of Sciences, Shanghai 201210, ChinaDepartment of Clinical Pharmacy, School of Pharmacy, Fudan University, Shanghai 201203, ChinaWarfarin is extensively used for venous thromboembolism and other coagulopathies. In clinical settings, warfarin is administered as a mixture of S- and R-warfarin, and both enantiomers are metabolized by multiple cytochrome P450 enzymes into many hydroxylation metabolites. Due to the high degree of structural similarity of hydroxylation metabolites, their profile possesses significant challenges. The previous methods generally suffer from lacking baseline resolution and/or involving complex analysis processes. To overcome this limitation, a sensitive and specific chiral liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed to simultaneously identify warfarin and hydroxywarfarins enantiomers. Chromatographic separation was achieved on a HYPERSIL CHIRAL-OT column. The mass spectrometric detection was carried out in negative ion MRM mode with electrospray ionization source. The optimized method exhibited satisfactory within-run and between-run accuracy and precision with lower limit of quantification (LLOQ) of 10.0 ng/mL and 1.0 ng/mL for warfarin and 7-, 10(R)-OH-warfarin enantiomers, respectively. Linear responses of warfarin enantiomers and 7-, and 10(R)-OH-warfarin enantiomers in rat plasma were observed over the range of 10.0–8000 ng/mL, and 1.00–800 ng/mL, respectively. The analytes were shown to be stable in various experimental conditions in rat plasma. Protein precipitation was used in sample preparation without a matrix effect. This method was successfully applied to pharmacokinetic study for quantitating the concentrations of S/R-warfarin, S/R-7-OH-warfarin, and S/R-10(R)-OH-warfarin and relatively quantitating 3′-, 4-, 6-, and 8-OH warfarin enantiomers in rat plasma.https://www.mdpi.com/1999-4923/14/6/1141enantiomershydroxywarfarinLC-MS/MSmetaboliteswarfarin |
| spellingShingle | Shasha Jin Zhihong Li Qing Yang Boyu Fang Xiaoqiang Xiang Chao Peng Weimin Cai Simultaneous Characterization and Determination of Warfarin and Its Hydroxylation Metabolites in Rat Plasma by Chiral Liquid Chromatography-Tandem Mass Spectrometry Pharmaceutics enantiomers hydroxywarfarin LC-MS/MS metabolites warfarin |
| title | Simultaneous Characterization and Determination of Warfarin and Its Hydroxylation Metabolites in Rat Plasma by Chiral Liquid Chromatography-Tandem Mass Spectrometry |
| title_full | Simultaneous Characterization and Determination of Warfarin and Its Hydroxylation Metabolites in Rat Plasma by Chiral Liquid Chromatography-Tandem Mass Spectrometry |
| title_fullStr | Simultaneous Characterization and Determination of Warfarin and Its Hydroxylation Metabolites in Rat Plasma by Chiral Liquid Chromatography-Tandem Mass Spectrometry |
| title_full_unstemmed | Simultaneous Characterization and Determination of Warfarin and Its Hydroxylation Metabolites in Rat Plasma by Chiral Liquid Chromatography-Tandem Mass Spectrometry |
| title_short | Simultaneous Characterization and Determination of Warfarin and Its Hydroxylation Metabolites in Rat Plasma by Chiral Liquid Chromatography-Tandem Mass Spectrometry |
| title_sort | simultaneous characterization and determination of warfarin and its hydroxylation metabolites in rat plasma by chiral liquid chromatography tandem mass spectrometry |
| topic | enantiomers hydroxywarfarin LC-MS/MS metabolites warfarin |
| url | https://www.mdpi.com/1999-4923/14/6/1141 |
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