Simultaneous Determination of Rivaroxaban and Enalapril in Rat Plasma by UPLC–MS/MS and Its Application to A Pharmacokinetic Interaction Study

  • Shu Zheng
  • Shun-Bin Luo
  • Yi-Bin Mei
  • Jing Guo
  • Li-Jun Tong
  • Qiang Zhang
  • Xia-Yan YeEmail author
Original Research Article


Background and Objectives

There have been no animal experiments and clinical studies on the pharmacokinetic interaction between rivaroxaban and enalapril. To investigate whether a potential pharmacokinetic interaction is present between rivaroxaban and enalapril, a rapid and sensitive Ultra performance liquid chromatography—tandem mass spectrometry (UPLC–MS/MS) method was developed and validated to determine the concentration of rivaroxaban and enalapril in rat plasma and was then applied to a pharmacokinetic interaction study.


The analytes were separated on an Acquity UPLC BEH C18 chromatography column (2.1 × 50 mm, 1.7 μm) with acetonitrile and 0.1% formic acid as the mobile phase with gradient elution. The mass spectrometer was operated in multiple reaction monitoring mode to monitor the precursor-to-product ion transitions of 436.1 → 145.1 m/z for rivaroxaban, 377.3 → 234.2 m/z for enalapril and 285.2 → 193.1 m/z for diazepam (IS).


The method was validated over the concentration range of 1.0–200 ng/mL for rivaroxaban and 0.5–100 ng/mL for enalapril. The intra- and inter-day precision and accuracy of the quality control (QC) samples exhibited relative standard deviations (RSD) < 9.4% and the accuracy values ranged from − 8.3 to 9.6%. After co-administration of rivaroxaban and enalapril, the maximum plasma concentration (Cmax) and area under the systemic drug concentration–time curve from time 0 to infinity (AUC0–∞) of rivaroxaban were significantly increased by 19.6% (p < 0.05) and 21.3% (p < 0.05), respectively. On the contrary, the plasma clearance rate (CL/F) of rivaroxaban and enalapril was significantly decreased by 17.8% (p < 0.05) and 23.8% (p < 0.05), respectively.


The UPLC–MS/MS method was successfully applied to simultaneous determination of rivaroxaban and enalapril in rat plasma and applied to study the pharmacokinetic interaction between rivaroxaban and enalapril. The co-administration of rivaroxaban and enalapril resulted in a significant drug interaction in rats.



The authors thank Guo-Xin Hu for providing DAS 2.0 software and his assistance and advice.

Compliance with Ethical Standards


No source of funding was used in this study.

Conflict of Interest

None of the authors has any potential conflicts of interest to declare.

Ethical Approval

All experiments were approved by the Wenzhou Medical University Committee, and all of the experiments were performed according to the National Institutes of Health Guide for the Care and Use of Laboratory Animals.


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Copyright information

© Springer Nature Switzerland AG 2018

Authors and Affiliations

  • Shu Zheng
    • 1
  • Shun-Bin Luo
    • 2
  • Yi-Bin Mei
    • 3
  • Jing Guo
    • 4
  • Li-Jun Tong
    • 3
  • Qiang Zhang
    • 5
  • Xia-Yan Ye
    • 6
    Email author
  1. 1.Department of Paediatric SurgeryThe People’s Hospital of LishuiLishuiChina
  2. 2.Department of Clinical PharmacyThe People’s Hospital of LishuiLishuiChina
  3. 3.Department of CardiologyThe People’s Hospital of LishuiLishuiChina
  4. 4.Department of Regional Medical UnionThe People’s Hospital of LishuiLishuiChina
  5. 5.Department of Clinical LaboratoryThe People’s Hospital of LishuiLishuiChina
  6. 6.Department of NeurosurgeryThe People’s Hospital of LishuiLishuiChina

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