Skip to main content
SpringerLink
Log in

A validated high-throughput UHPLC-MS/MS assay for accurate determination of rivaroxaban in plasma sample

  • Published:
Journal of Thrombosis and Thrombolysis Aims and scope Submit manuscript

Abstract

Rivaroxaban is a novel, selective and potent oral direct factor Xa inhibitor, therapeutically indicated in the treatment of thromboembolic diseases. Like traditional anticoagulants, routine coagulation monitoring of rivaroxaban is not necessary, but important in some clinical circumstances. In this study, a sensitive UHPLC-MS/MS assay for rapid determination of rivaroxaban in human plasma was developed and validated. Rivaroxaban and its internal standard (IS) were extracted from plasma using acetonitrile as protein precipitating agent. An isocratic mobile phase of acetonitrile: 10 mM ammonium acetate (80:20, v/v) at a flow rate of 0.3 mL/min was used for the separation of rivaroxaban and IS. Both rivaroxaban and IS was eluted within 1 min with a total run time of 1.5 min only. Electrospray ionization source in positive mode was used for the detections of rivaroxaban and IS. Precursor to product ion transition of m/z 436.00 > 144.87 for rivaroxaban and m/z 411.18 > 191.07 for IS were used in multiple reaction monitoring mode. Developed assay was fully validated in terms of selectivity, linearity, accuracy, precision, recovery, matrix effects and stability using official guideline on bioanalytical method.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Mann KG, Brummel K, Butenas S (2003) What is all that thrombin for? J Thromb Haemost 1:1504–1514

    Article  CAS  PubMed  Google Scholar 

  2. Turpie AG (2007) Oral, direct factor Xa inhibitors in development for the prevention and treatment of thromboembolic diseases. Arterioscler Thromb Vasc Biol 27:1238–1247

    Article  CAS  PubMed  Google Scholar 

  3. Kreutz R (2012) Pharmacodynamic and pharmacokinetic basics of rivaroxaban. Fund Clin Pharmacol 26:27–32

    Article  CAS  Google Scholar 

  4. Lindhoff-Last E, Samama MM, Ortel TL, Weitz JI, Spiro TE (2010) Assays for measuring rivaroxaban: their suitability and limitations. Ther Drug Monit 32:673–679

    Article  CAS  PubMed  Google Scholar 

  5. Sarich TC, Peters G, Berkowitz SD, Misselwitz F, Nessel CC, Burton P et al (2013) Rivaroxaban: a novel oral anticoagulant for the prevention and treatment of several thrombosis-mediated conditions. Ann N Y Acad Sci 1291:42–55

    Article  CAS  PubMed  Google Scholar 

  6. Kubitza D, Becka M, Voith B, Zuehlsdorf M, Wensing G (2005) Safety, pharmacodynamics, and pharmacokinetics of single doses of BAY 59-7939, an oral, direct factor Xa inhibitor. Clin Pharmacol Ther 78:412–421

    Article  CAS  PubMed  Google Scholar 

  7. Kubitza D, Becka M, Wensing G, Voith B, Zuehlsdorf M (2005) Safety, pharmacodynamics, and pharmacokinetics of BAY 59-7939–an oral, direct Factor Xa inhibitor–after multiple dosing in healthy male subjects. Eur J Clin Pharmacol 61:873–880

    Article  CAS  PubMed  Google Scholar 

  8. Mueck W, Stampfuss J, Kubitza D, Becka M (2014) Clinical pharmacokinetic and pharmacodynamic profile of rivaroxaban. Clin Pharmacokinet 53:1–16

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  9. Kreutz R (2014) Pharmacokinetics and pharmacodynamics of rivaroxaban–an oral, direct factor Xa inhibitor. Curr Clin Pharmacol 9:75–83

    Article  CAS  PubMed  Google Scholar 

  10. Mueck W, Schwers S, Stampfuss J (2013) Rivaroxaban and other novel oral anticoagulants: pharmacokinetics in healthy subjects, specific patient populations and relevance of coagulation monitoring. Thromb J 11:10

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  11. Risselada AJ, Visser MJ, van Roon E (2013) Pulmonary embolism due to interaction between rivaroxaban and carbamazepine. Ned tijdschrift voor geneeskunde 157:A6568

    Google Scholar 

  12. Douxfils J, Tamigniau A, Chatelain B, Chatelain C, Wallemacq P, Dogne JM et al (2013) Comparison of calibrated chromogenic anti-Xa assay and PT tests with LC-MS/MS for the therapeutic monitoring of patients treated with rivaroxaban. Thromb Haemost 110:723–731

    Article  CAS  PubMed  Google Scholar 

  13. Harenberg J, Erdle S, Marx S, Kramer R (2012) Determination of rivaroxaban in human plasma samples. Semin Thromb Hemost 38:178–184

    Article  CAS  PubMed  Google Scholar 

  14. Rohde G (2008) Determination of rivaroxaban–a novel, oral, direct Factor Xa inhibitor–in human plasma by high-performance liquid chromatography-tandem mass spectrometry. J Chromatog B 872:43–50

    Article  CAS  Google Scholar 

  15. Gous T, Couchman L, Patel JP, Paradzai C, Arya R, Flanagan RJ (2014) Measurement of the Direct Oral Anticoagulants Apixaban, Dabigatran, Edoxaban, and Rivaroxaban in Human Plasma Using Turbulent Flow Liquid Chromatography With High-Resolution Mass Spectrometry. Ther Drug Monit. doi:10.1097/FTD.0000000000000059.

  16. Magiera S (2013) Fast, simultaneous quantification of three novel cardiac drugs in human urine by MEPS-UHPLC-MS/MS for therapeutic drug monitoring. J Chromatog B 938:86–95

    Article  CAS  Google Scholar 

  17. US Food and Drug Administration (2001) Guidance for Industry on Bioanalytical Method Validation. Center for Drug Evaluation and Research (CDER). US Food and Drug Administration, Rockville

    Google Scholar 

  18. European Medicines Agency (2011) Guideline on bioanalytical method validation. CHMP. European Medicines Agency, London

    Google Scholar 

  19. Samama MM, Contant G, Spiro TE, Perzborn E, Flem LL, Guinet C et al (2012) Evaluation of the prothrombin time for measuring rivaroxaban plasma concentrations using calibrators and controls: results of a multicenter field trial. Clin Appl Thromb Hemost 18:150–158

    Article  CAS  PubMed  Google Scholar 

  20. Samama MM, Contant G, Spiro TE, Perzborn E, Guinet C, Gourmelin Y et al (2012) Evaluation of the anti-factor Xa chromogenic assay for the measurement of rivaroxaban plasma concentrations using calibrators and controls. Thromb Haemost 107:379–387

    Article  CAS  PubMed  Google Scholar 

  21. Mani H, Rohde G, Stratmann G, Hesse C, Herth N, Schwers S et al (2012) Accurate determination of rivaroxaban levels requires different calibrator sets but not addition of antithrombin. Thromb Haemost 108:191–198

    Article  CAS  PubMed  Google Scholar 

  22. Lippi G, Ardissino D, Quintavalla R, Cervellin G (2014) Urgent monitoring of direct oral anticoagulants in patients with atrial fibrillation: a tentative approach based on routine laboratory tests. J Thromb Thrombolyis 38(2):269–274

    Article  CAS  Google Scholar 

  23. Liu G, Snapp HM, Ji QC, Arnold ME (2009) Strategy of accelerated method development for high-throughput bioanalytical assays using ultra high-performance liquid chromatography coupled with mass spectrometry. Anal Chem 81:9225–9232

    Article  CAS  PubMed  Google Scholar 

  24. Kumar A, Saini G, Nair A, Sharma R (2012) UPLC: a preeminent technique in pharmaceutical analysis. Acta Poly Pharm 69:371–380

    CAS  Google Scholar 

  25. Zheng N, Zeng J, Akinsanya B, Buzescu A, Xia YQ, Ly V et al (2013) A rapid, accurate and robust UHPLC-MS/MS method for quantitative determination of BMS-927711, a CGRP receptor antagonist, in plasma in support of non-clinical toxicokinetic studies. J Pharm Biomed Anal 283:237–248

    Article  Google Scholar 

Download references

Acknowledgments

The authors extend their appreciation to the Deanship of Scientific Research at King Saud University for funding the work through the research group project No. RGP-VPP-203.

Author information

Authors and Affiliations

  1. Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, PO BOX 2457, Riyadh, 11451, Saudi Arabia

    Muzaffar Iqbal & Nasr Y. Khalil

  2. Bioavailability Laboratory, College of Pharmacy, King Saud University, Riyadh, 11451, Saudi Arabia

    Muzaffar Iqbal

  3. Department of Pharmacology and Toxicology, College of Pharmacy, King Saud University, Riyadh, 11451, Saudi Arabia

    Faisal Imam

  4. Department of Pharmaceutics, College of Pharmacy, Salman bin Abdulaziz University, Al-Kharj, Saudi Arabia

    Md. Khalid Anwer

Authors
  1. Muzaffar Iqbal
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nasr Y. Khalil
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Faisal Imam
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Md. Khalid Anwer
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Muzaffar Iqbal.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Iqbal, M., Khalil, N.Y., Imam, F. et al. A validated high-throughput UHPLC-MS/MS assay for accurate determination of rivaroxaban in plasma sample. J Thromb Thrombolysis 39, 79–88 (2015). https://doi.org/10.1007/s11239-014-1121-2

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11239-014-1121-2

Keywords

Search

Navigation