Skip to main content
SpringerLink
Log in

A Critical N-Nitrosamine Impurity of Anticoagulant Drug, Rivaroxaban: Synthesis, Characterization, Development of LC–MS/MS Method for Nanogram Level Quantification

  • Original
  • Published:
Chromatographia Aims and scope Submit manuscript

Abstract

Background and Purpose

The product recall for detecting unacceptable levels of carcinogenic potential N-nitrosamine impurities has been a severe apprehension in recent years. The regulatory agencies issued guidelines for the pharmaceutical industries to monitor nitrosamine impurities in the drug products. In the present study, we have been developed a robust, simple and sensitive ultra-high-performance liquid chromatography (UHPLC) coupled with mass spectrometry (LC–MS/MS) method for the determination of possible N-nitrosamine impurity, N-(2-hydroxyethyl)-N-phenylnitrous amide in the drug molecule, Rivaroxaban at nanogram level.

Methods

Chromatographic experiments were optimized on VD-Spher100 C18 E (150 mm × 4.6 mm, 3 μm) column, mobile phase flow was operated in isocratic mode using 0.1% aqueous formic acid and methanol in the ratio of 1:1 (v/v) with a flow rate of 0.6 mL/min, and column oven temperature maintained at 40 °C. The developed method was validated by following Q2(R1) International Conference on Harmonization” (ICH) guidelines.

Results

The limit of detection (LOD) and quantification (LOQ) of this N-nitrosamine impurity was 0.045 ng mL−1 and 0.15 ng mL−1, respectively. The accuracy and precision of the method were found well within the specified range. In addition, the synthesis and characterization details of N-nitrosamine impurity, N-(2-hydroxyethyl)-N-phenylnitrous amide were provided.

Conclusions

The developed method was applied well to determine N-nitrosamine impurity in bulk-scale samples of Rivaroxaban, revealing that the method could be worthwhile to use in the analysis of pharmaceutical dosage forms.

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
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Gyorgy S, Miriam C, Amores D, Marco G, Frederico CF, William H (2015) Genotoxic impurities in pharmaceutical manufacturing: sources, regulations, and mitigation. Chem Rev 115:8182–8229. https://doi.org/10.1021/cr300095f

    Article  CAS  Google Scholar 

  2. Linda KD, Marvin MH, Brian WP, Todd JP, Steven WB (2013) The assessment of impurities for genotoxic potential and subsequent control in drug substance and drug product. J Pharm Sci 102:1404–1418. https://doi.org/10.1002/jps.23462

    Article  CAS  Google Scholar 

  3. Amit G, Hussain S, Tabrez S (2018) Genotoxic impurities and its risk assessment in drug compound. DDIPIJ. https://doi.org/10.32474/DDIPIJ.2018.02.000143

    Article  Google Scholar 

  4. Rene H, David PE (2016) Analytical advances in pharmaceutical impurity profiling. Eur J Pharm Sci 87:118–135. https://doi.org/10.1016/j.ejps.2015.12.007

    Article  CAS  Google Scholar 

  5. De la Monte SM, Tong M (2009) Mechanisms of nitrosamine-mediated neurodegeneration: potential relevance to sporadic Alzheimer’s disease. J Alzheimer’s Dis 17(4):817–825. https://doi.org/10.3233/jad-2009-1098

    Article  Google Scholar 

  6. McGovern T, Jacobson-Kram D (2006) Regulation of genotoxic and carcinogenic impurities in drug substances and products. TrAC Trends Anal Chem 25:790–795. https://doi.org/10.1016/j.trac.2006.06.004

    Article  CAS  Google Scholar 

  7. Sonali SB (2021) Critical analysis of drug product recalls due to nitrosamine impurities. J Med Chem 64:2923–2936. https://doi.org/10.1021/acs.jmedchem.0c02120

    Article  CAS  Google Scholar 

  8. Anonymous (2008) Guidance (Draft) for industry genotoxic and carcinogenic impurities in drug substances and products: Recommended approaches. Center for Drug Evaluation and Research, Food and Drug Administration

  9. Challis BC (1985) Nutrition and nitrosamine formation. Proc Nutr Soc 44:95–100. https://doi.org/10.1079/PNS19850015

    Article  CAS  PubMed  Google Scholar 

  10. FDA-CDER (2020) Control of nitrosamine impurities in human drugs guidance for industry. Docket Number: FDA-2020-D-1530. https://www.fda.gov/regulatory-information/search-fda-guidance-documents/control-nitrosamine-impurities-human-drugs

  11. Gushgari AJ, Halden RU (2018) Critical review of major sources of human exposure to N-nitrosamines. Chemosphere 210:1124–1136. https://doi.org/10.1016/j.chemosphere.2018.07.098

    Article  CAS  PubMed  Google Scholar 

  12. https://www.fda.gov/drugs/drug-safety-and-availability/fda-updates-and-pressannouncements-ndma-metformin. Accessed 10 May 2020

  13. European pharmacopeia 10.6 chapters (2.5.42). N-Nitrosamines in active substances and revised Sartan monographs. https://www.edqm.eu/en/news/general-chapter-2542-n-nitrosamines-active-substances-and-revised-sartan-monographs

  14. EMA, ICH guideline M7(R1) on assessment and control of DNA reactive (mutagenic) impurities in pharmaceuticals to limit potential carcinogenic risk, 25 August 2015: EMA/CHMP/ICH/83812/2013

  15. FDA, CDER, M7 (R1) (2018) Assessment and control of DNA reactive (mutagenic) impurities in pharmaceuticals to limit potential carcinogenic risk guidance for industry. U.S. Department of Health and Human Services

  16. International Conference on Harmonization (ICH) (2017). Guideline M7 (R1) Assessment and control of DNA reactive (mutagenic) impurities in pharmaceuticals to limit potential carcinogenic risk. March 2017

  17. EMA (2021) Questions and answers for marketing authorisation holders/applicants on the CHMP Opinion for the Article 5(3) of Regulation (EC) No 726/2004 referral on nitrosamine impurities in human medicinal products, EMA/409815/2020 Rev.6

  18. Becker R, Berkowitz SD, Breithardt G (2010) Rivaroxaban-once daily, oral, direct factor Xa inhibition compared with vitamin K antagonism for prevention of stroke and Embolism Trial in Atrial Fibrillation: rationale and design of the ROCKET AF study. Am Heart J 159:340–347. https://doi.org/10.1016/j.ahj.2009.11.025

    Article  CAS  Google Scholar 

  19. Roehrig S, Straub A, Pohlmann J, Lampe T, Pernerstorfer J, Schlemmer KH, Reinemer P, Perzborn E (2005) Discovery of the novel antithrombotic agent 5-chloro-N-({(5S)-2-oxo-3-[4-(3-oxomorpholin-4-yl)phenyl]-1,3-oxazolidin-5-yl}methyl)thiophene-2-carboxamide (BAY 59–7939): an oral, direct factor Xa inhibitor. J Med Chem 48:5900–5908. https://doi.org/10.1021/jm050101d

    Article  CAS  PubMed  Google Scholar 

  20. Mann KG, Brummel K, Butenas S (2003) What is all that thrombin for? J Thromb Haemost 1:1504–1514. https://doi.org/10.1046/j.1538-7836.2003.00298.x

    Article  CAS  PubMed  Google Scholar 

  21. 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. Thrombosis J. https://doi.org/10.1186/1477-9560-11-10

    Article  Google Scholar 

  22. Roehrig S, Straub A, Pohlmann J, Lampe T, Pernerstorfer J, Schlemmer KH (2005) Discovery of the novel antithrombotic agent 5-chloro-n-({(5s)-2- oxo-3- [4-(3-oxomorpholin-4-yl)phenyl]-1,3- oxazolidin-5-yl}methyl)thiophene- 2-carboxamide (bay 59–7939): an oral, direct factor xa inhibitor. J Med Chem 48:5900–5908. https://doi.org/10.1021/jm050101d

    Article  CAS  PubMed  Google Scholar 

  23. Biemond BJ, Perzborn E, Friederich PW, Levi M, Buetehorn U, Buller HR (2007) Prevention and treatment of experimental thrombosis in rabbits with rivaroxaban (BAY 597939)-an oral, direct factor Xa inhibitor. Thromb Haemost 97:471–477. https://doi.org/10.1160/TH06-11-0620

    Article  CAS  PubMed  Google Scholar 

  24. Gulseth MP, Michaud J, Nutescu EA (2008) Rivaroxaban: an oral direct inhibitor of factor Xa. Am J Health-Syst Ph 65:1520–1529. https://doi.org/10.2146/ajhp070624

    Article  CAS  Google Scholar 

  25. Global Rivaroxaban Market Growth at a CAGR of 8.2% during 2018–2025. https://express-press-release.net/news/2018/06/19/275877. Accessed 19 June 2018

  26. Tanzeela Abdul F, Aamer S (2017) A review on the synthetic approaches of rivaroxaban an anticoagulant drug. Tetrahedron Asymmetry 28:485–504. https://doi.org/10.1016/j.tetasy.2017.02.010

    Article  CAS  Google Scholar 

  27. USP General Chapter <1225> Validation of compendial procedures

  28. ICH Harmonised Tripartite Guideline, validation of analytical procedures. Text and methodology Q2(R1). In: International conference on harmonisation of technical requirements for registration of pharmaceuticals for human use. 2005

  29. Food and Drug Administration Office of Regulatory Affairs ORA Laboratory Manual Volume II (2020) Methods, method verification and validation, Document Number: ORA-LAB.5.4.5

  30. Zerong W (2010) Sandmeyer reaction. Comprehensive Organic name reactions and reagents. Wiley, Hoboken

    Google Scholar 

Download references

Acknowledgements

The authors are obliged to the management of Micro Labs Ltd., API Division Centre, ML-27, Bangalore, for providing all the test samples, reference standards, solvents, reagents, and lab facilities to carry out the present research work. We also wish to thank Dr. G. Madhusudana Rao, Bhattacharyya Gouri, and Surya Krishna Mohan for their valuable support and cooperation.

Author information

Authors and Affiliations

  1. Analytical Research and Development, Micro Labs Ltd, API R&D Centre, Plot No. 43-45, 4th Phase, KIADB, JB Link Road, Bengaluru, 560105, Karnataka, India

    Vijaya Kumar Baksam, Subba Rao Devineni, Mohit Jain, Prashanth Kumar, Sanjeev Shandilya & Pramod Kumar

  2. Department of Chemistry, JNTUA College of Engineering, Kalikiri, Chittoor, 576213, Andhra Pradesh, India

    Vijaya Kumar Baksam & Nimmakayala Saritha

Authors
  1. Vijaya Kumar Baksam
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nimmakayala Saritha
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Subba Rao Devineni
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mohit Jain
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Prashanth Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Sanjeev Shandilya
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Pramod Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Vijaya Kumar Baksam.

Ethics declarations

Conflict of interest

The authors declare no conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 506 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Baksam, V.K., Saritha, N., Devineni, S.R. et al. A Critical N-Nitrosamine Impurity of Anticoagulant Drug, Rivaroxaban: Synthesis, Characterization, Development of LC–MS/MS Method for Nanogram Level Quantification. Chromatographia 85, 73–82 (2022). https://doi.org/10.1007/s10337-021-04115-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10337-021-04115-x

Keywords

Search

Navigation