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Related Experiment Videos

Simple and sensitive high-performance liquid chromatographic.

K Na-Bangchang1, O Supasyndh, T Supaporn

  • 1Clinical Pharmacology Unit, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand. tmknb@mucc.mahidol.ac.th

Journal of Chromatography. B, Biomedical Sciences and Applications
|April 25, 2000
PubMed
Summary

This study presents a rapid, selective, and sensitive HPLC method for quantifying mycophenolic acid (MPA), a key metabolite of the immunosuppressant mycophenolate mofetil (MMF). The developed assay is suitable for clinical pharmacokinetic studies.

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Area of Science:

  • Analytical Chemistry
  • Pharmacokinetics
  • Clinical Chemistry

Background:

  • Mycophenolic acid (MPA) is the active metabolite of mycophenolate mofetil (MMF), an important immunosuppressant drug.
  • Accurate quantification of MPA in plasma is crucial for therapeutic drug monitoring and pharmacokinetic studies.

Purpose of the Study:

  • To develop and validate a rapid, selective, sensitive, and reproducible reversed-phase High-Performance Liquid Chromatography (HPLC) method for the quantitative determination of MPA in human plasma.
  • To assess the suitability of the developed method for clinical pharmacokinetic investigations.

Main Methods:

  • A one-step liquid-liquid extraction using dichloromethane-dichloroethane at acidic pH was employed for sample preparation.
  • Chromatographic separation was achieved using a C18 column with a mobile phase consisting of acetonitrile and a phosphate buffer.

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  • Detection of MPA was performed using UV detection at 254 nm.

    • Main Results:

    • The method demonstrated high mean recoveries for MPA (89-98%) and the internal standard (90-96%).
    • Excellent precision was observed with within-day coefficients of variation ranging from 0.3-7.8% and day-to-day variations of 1.1-2.0%.
    • The minimum detectable concentration for MPA in plasma was as low as 0.005 microg/ml.

    Conclusions:

    • The developed reversed-phase HPLC method is rapid, selective, sensitive, and reproducible for MPA quantification in plasma.
    • This validated method is suitable for application in clinical pharmacokinetic studies of mycophenolate mofetil.