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

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. © 2019 Elsevier B.V.

Authors
Kozlov A.V. 1 , Ramenskaya G.V.1 , Sychev D.A.2 , Vlasov A.M.1 , Makarenkova L.M. 3 , Stepanova E.S. 3 , Gegechkori V.I.1 , Agatonovic-Kustrin S.1 , Chistyakov V.V.I. 3
Publisher
Elsevier B.V.
Language
English
Status
Published
Number
121808
Volume
1130-1131
Year
2019
Organizations
  • 1 Department of Pharmaceutical and Toxicological Chemistry, I. M. Sechenov First Moscow State Medical University (Sechenov University), Moscow, 119991, Russian Federation
  • 2 FGBOU DPO RMAPO Ministry of Health of Russia, Department of Clinical Pharmacology and Therapy, Educational and Laboratory Building of RMAPO, Moscow, 125993, Russian Federation
  • 3 Federal State Autonomous Educational Institution of Higher Professional Education Peoples Friendship University of Russia “The Center for Collective Use” (Scientific-Educational Center), Moscow, 117198, Russian Federation
Keywords
Anticoagulant; Dabigatran; Extraction protocol; HPLC-MS/MS; Pharmacokinetics
Date of creation
24.12.2019
Date of change
24.12.2019
Short link
https://repository.rudn.ru/en/records/article/record/54925/
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