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Related Experiment Videos

Separation methods for methotrexate, its structural analogues and metabolites.

F M Rubino1

  • 1Department of Medicine, Surgery and Odontology, University of Milano at Ospedale S. Paolo, Milan, Italy. federico.rubino@unimi.it

Journal of Chromatography. B, Biomedical Sciences and Applications
|January 31, 2002
PubMed
Summary

This review details chromatographic methods for measuring methotrexate (MTX) and its metabolites in biological samples. It covers various techniques for therapeutic drug monitoring and research, aiding antifolate drug discovery.

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Area of Science:

  • Analytical Chemistry
  • Pharmacology
  • Biochemistry

Background:

  • Methotrexate (MTX) is a crucial antifolate drug used in cancer, immunosuppression, and autoimmune disease treatment.
  • Accurate measurement of MTX and its metabolites is vital for therapeutic drug monitoring and fundamental research.
  • Chromatographic methods are established techniques for quantifying MTX in various biological matrices.

Purpose of the Study:

  • To review and summarize published chromatographic methods for measuring MTX, its metabolites, and analogues.
  • To assess the instrumental conditions and performance of these chromatographic assays.
  • To highlight the importance of sensitive and efficient methods for clinical and research applications.

Main Methods:

  • Review of over 70 published papers on chromatographic assays for MTX and metabolites (1975-2000).

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  • Analysis of diverse sample preparation, analyte separation (various stationary and mobile phases), and detection techniques (UV-Vis, fluorimetry, electrochemistry, MS).
  • Discussion of methods for analyzing MTX and metabolites in plasma, serum, saliva, urine, CSF, and tissues.

    • Main Results:

    • A wide array of chromatographic conditions have been employed for MTX and metabolite analysis.
    • Optimized methods enable rapid, simultaneous measurement of MTX and main metabolites in plasma at low-nM concentrations.
    • Chromatographic analysis is crucial for workplace contamination monitoring and basic research on folate metabolism.

    Conclusions:

    • Chromatographic techniques offer versatile and effective means for quantifying MTX and its metabolites.
    • The development of sensitive, fast, and cost-effective methods is essential for clinical practice and research.
    • Chromatography aids in identifying metabolites and monitoring environmental and biological contamination.