Skip to main content
SpringerLink
Log in

UPLC-MS/MS Analysis of Methotrexate in Human Plasma and Comparison with the Fluorescence Polarization Immunoassay

  • Published:
Analytical Sciences Aims and scope Submit manuscript

Abstract

Methotrexate (MTX) plasma concentration is routinely monitored to guide the dosage regimen of rescue drugs. This study aims to develop and validate an ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for plasma MTX analysis, and to establish its agreement with the fluorescence polarization immunoassay (FPIA) in patients with high-dose MTX therapy. Separation was achieved by gradient elution with methanol and water (0.05% formic acid) at 40°C with a run time of 3 min. The intra- and inter-day inaccuracy and imprecision of the UPLC-MS/MS method were -4.25 to 3.1 and less than 7.63%, respectively. The IS-normalized recovery and matrix effect were 87.05 to 92.81 and 124.43 to 134.57%. The correlation coefficients between UPLC-MS/MS and FPIA were greater than 0.98. The UPLC-MS/MS method was in agreement with the FPIA at high levels of MTX (1.0 - 100 μmol/L), but not at low levels (0.01 - 1.0 μmol/L). Further studies are warranted to confirm these results.

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

Similar content being viewed by others

References

  1. Hospira, Label for Methotrexate Injection, 2011, http://www.accessdata.fda.gov/drugsatfda_docs/label/2011/011719s117lbl.pdf, Accessed May 19, 2012.

  2. M. A. Pesce and S. H. Bodourian, Ther. Drug Monit., 1986, 8, 115.

    Article  CAS  PubMed  Google Scholar 

  3. K. Michail and M. S. Moneeb, J. Pharm. Biomed. Anal., 2011, 55, 317.

    Article  CAS  PubMed  Google Scholar 

  4. D. Wu, Y. Wang, Y. Sun, N. Ouyang, and J. Qian, Biomed. Chromatogr., 2015, 29, 1197.

    Article  CAS  PubMed  Google Scholar 

  5. R. Wang, L. Guo, H. Xie, J. Zhang, X. Li, W. Li, J. Wang, X. Wu, and Z. Jia, Cell Biochem. Biophys., 2013, 67, 1343.

    Article  PubMed  Google Scholar 

  6. I. Rodin, A. Braun, A. Stavrianidi, and O. Shpigun, J. Chromatogr. B, 2013, 937, 1.

    Article  CAS  Google Scholar 

  7. S. R. Thappali, K. V. Varanasi, S. Veeraraghavan, S. K. Vakkalanka, and M. Khagga, Arzneim. Forsch., 2012, 62, 624.

    Article  CAS  Google Scholar 

  8. E. Sonemoto, N. Kono, R. Ikeda, M. Wada, Y. Ueki, and K. Nakashima, Biomed. Chromatogr., 2012, 26, 1297.

    Article  CAS  PubMed  Google Scholar 

  9. X. Mo, Y. Wen, B. Ren, J. Chen, C. Xu, X. Wang, M. Huang, and X. Chen, J. Chromatogr. B, 2012, 907, 41.

    Article  CAS  Google Scholar 

  10. G. Chen, J. P. Fawcett, M. Mikov, and I. G. Tucker, J. Pharm. Biomed. Anal., 2009, 50, 262.

    Article  CAS  PubMed  Google Scholar 

  11. P. Guo, X. Wang, L. Liu, M. G. Belinsky, G. D. Kruh, and J. M. Gallo, J. Pharm. Biomed. Anal., 2007, 43, 1789.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. A. Barbieri, L. Sabatini, P. Indiveri, R. Bonfiglioli, V. Lodi, and F. S. Violante, Rapid Commun. Mass Spectrom., 2006, 20, 1889.

    Article  CAS  PubMed  Google Scholar 

  13. Y. Suzuki, H. Arakawa, and M. Maeda, Anal. Sci., 2003, 19, 111.

    Article  CAS  PubMed  Google Scholar 

  14. R. Bouquie, M. Gregoire, H. Hernando, C. Azoulay, E. Dailly, C. Monteil-Ganiere, A. Pineau, G. Deslandes, and P. Jolliet, Am. J. Clin. Pathol., 2016, 146, 119.

    Article  CAS  PubMed  Google Scholar 

  15. F. J. Primus, E. A. Kelley, H. J. Hansen, and D. M. Goldenberg, Clin. Chem., 1988, 34, 261.

    Article  CAS  PubMed  Google Scholar 

  16. S. Buchen, D. Ngampolo, R. G. Melton, C. Hasan, A. Zoubek, G. Henze, U. Bode, and G. Fleischhack, Br. J. Cancer, 2005, 92, 480.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. E. den Boer, B. C. Koch, R. Huisman, and R. de Jonge, Ther. Drug Monit., 2014, 36, 819.

    Article  Google Scholar 

  18. M. R. Al-Turkmani, T. Law, A. Narla, and M. D. Kellogg, Clin. Chem., 2010, 56, 1792.

    Article  CAS  PubMed  Google Scholar 

  19. E. den Boer, S. G. Heil, B. D. van Zelst, R. J. Meesters, B. C. Koch, M. L. Te Winkel, M. M. van den Heuvel-Eibrink, T. M. Luider, and R. de Jonge, Ther. Drug Monit., 2012, 34, 432.

    Article  Google Scholar 

  20. J. M. Bland and D. G. Altman, Stat. Methods Med. Res., 1999, 8, 135.

    Article  CAS  PubMed  Google Scholar 

  21. J. M. Bland and D. G. Altman, Lancet, 1986, 1, 307.

    Article  CAS  PubMed  Google Scholar 

  22. CLSI, Mass Spectrometry in the Clinical Laboratory: General Principles and Guidance; Approved Guideline, 2007, http://shop.clsi.org/c.1253739/site/Sample_pdf/C50A_sample.pdf, Accessed Oct. 06, 2011.

    Google Scholar 

  23. FDA, “Guidance for Industry: Bioanalytical Method Validation”, 2013, http://www.fda.gov/ucm/groups/fdagov-public/@fdagov-drugs-gen/documents/document/ucm368107.pdf, Accessed July 21, 2014.

    Google Scholar 

  24. H. Passing and Bablok, J. Clin. Chem. Clin. Biochem., 1983, 21, 709.

    CAS  PubMed  Google Scholar 

  25. J. M. Bland, “How Can I Decide the Sample Size for a Study of Agreement between Two Methods of Measurement?”, 2004, http://www-users.york.ac.uk/~mb55/meas/sizemeth.htm, Accessed Dec. 15, 2013.

    Google Scholar 

Download references

Acknowledgments

Thanks are given to our patients and our whole team. The authors would like to acknowledge Drs. Jiawang Liu, Ming Zhao and Shiqi Peng, College of Pharmaceutical Science, Capital Medical University, Beijing, China, for their contribution in revising the manuscript. This work was supported by Capital Medical University (No. 16JL13).

Author information

Authors and Affiliations

  1. Department of Pharmacy, Beijing Tiantan Hospital, Capital Medical University, 6 Tiantan Xili, Dongcheng District, Beijing, 100050, P. R. China

    Shenghui Mei, Leting Zhu, Xingang Li, Jiaqing Wang, Jiping Huo, Li Yang & Zhigang Zhao

  2. Department of Clinical Pharmacology, College of Pharmaceutical Sciences, Capital Medical University, Beijing, 100045, P. R. China

    Shenghui Mei, Xingang Li, Xueyun Jiang, Haiyan Chen, Li Yang & Zhigang Zhao

  3. Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, 6 Tiantan Xili, Dongcheng District, Beijing, 100050, P. R. China

    Song Lin

Authors
  1. Shenghui Mei
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Leting Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xingang Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jiaqing Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Xueyun Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Haiyan Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jiping Huo
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Li Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Song Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Zhigang Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Song Lin or Zhigang Zhao.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Mei, S., Zhu, L., Li, X. et al. UPLC-MS/MS Analysis of Methotrexate in Human Plasma and Comparison with the Fluorescence Polarization Immunoassay. ANAL. SCI. 33, 665–670 (2017). https://doi.org/10.2116/analsci.33.665

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.2116/analsci.33.665

Keywords

Search

Navigation