Репозиторий Университета

An improved extraction protocol for therapeutic dabigatran monitoring using HPLC-MS/MS


  • Kozlov A.
  • Ramenskaya G.
  • Sychev D.
  • Vlasov A.
  • Makarenkova L.
  • Stepanova E.
  • Gegechkori V.
  • Agatonovic-Kustrin S.
  • Chistyakov V.
Дата публикации:01.11.2019
Журнал: Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
БД: SCOPUS
Ссылка: SCOPUS

Аннтотация

© 2019 Elsevier B.V. A new sample extraction protocol was developed for pharmacokinetic studies of dabigatran with high-performance liquid chromatography separation - electrospray ionization time-of-flight mass spectrometry analysis. After protein precipitation with acetonitrile, free dabigatran and its metabolites are separated into water phase by water-dichloromethane liquid-liquid extraction to purify the sample from proteins and endogenous lipophilic compounds. Chromatographic separation was achieved on an Agilent Zorbax SB-CN column (150 × 4.6 mm, 5 µm)) using 0.1% aqueous solution of formic acid and acetonitrile (80:20) as the mobile phase. Agilent Zorbax SB-CN column was selected to improve sample resolution and to avoided early elution of dabigatran previously seen when using a C18 column. The extended calibration curve was constructed from 5 to 1000 ng/L while precision and accuracy were assessed at four levels across the linear dynamic ranges. Within-run precision was <5.6% and the between-run precision was <3.9%. The method accuracy ranged from 89.8% to 104.4%. The developed method was successfully applied to 30 patient samples to evaluate antithrombotic efficacy and anticoagulant activity of dabigatran following knee endoprosthesis surgery.


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