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Related Experiment Videos

Competitive protein binding assay for piritrexim.

J L Woolley1, J L Ringstad, C W Sigel

  • 1Wellcome Research Laboratories, Research Triangle Park, NC 27709.

Journal of Pharmaceutical Sciences
|September 1, 1989
PubMed
Summary
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A new radioassay accurately measures piritrexim (PTX) in plasma by utilizing a methotrexate kit. This competitive protein binding assay offers a sensitive and reliable method for PTX quantification.

Area of Science:

  • Pharmacology
  • Analytical Chemistry
  • Biochemistry

Background:

  • Piritrexim (PTX) is an antifolate drug.
  • Accurate quantification of PTX in biological samples is crucial for therapeutic drug monitoring.
  • Existing analytical methods may have limitations in sensitivity or accessibility.

Purpose of the Study:

  • To develop and validate a competitive protein binding assay for the quantification of piritrexim (PTX) in human plasma.
  • To adapt a commercially available radioassay kit for methotrexate for PTX measurement.
  • To establish the sensitivity, range, and precision of the developed assay.

Main Methods:

  • A competitive protein binding assay was developed using a commercially available radioassay kit for methotrexate.
  • Piritrexim (PTX) was selectively extracted from plasma.

Related Experiment Videos

  • PTX competed with [125I]methotrexate for binding to dihydrofolate reductase from Lactobacillus casei.
  • Free drug was separated from bound drug using dextran-coated charcoal adsorption.

    • Main Results:

    • The assay accurately quantifies piritrexim (PTX) over a concentration range of 0.01 to 10.0 micrograms/mL in plasma.
    • The assay demonstrated a coefficient of variation of less than 15%.
    • The limit of sensitivity for piritrexim (PTX) was approximately 2 ng/mL, and it correlated well with a previously published HPLC method.

    Conclusions:

    • A sensitive and reliable radioassay method for piritrexim (PTX) in plasma has been successfully developed.
    • This assay utilizes readily available reagents and offers a viable alternative to existing methods.
    • The assay is suitable for therapeutic drug monitoring of piritrexim (PTX).