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A rapid enzymatic assay for methotrexate in serum
Journal of Clinical Chemistry and Clinical Biochemistry. Zeitschrift Fur Klinische Chemie Und Klinische Biochemie
|April 1, 1979
Summary
A new kinetic assay accurately measures serum methotrexate levels within 1.5 hours. This sensitive method offers reliable results comparable to existing techniques for methotrexate monitoring.
Area of Science:
- Biochemistry
- Enzymology
- Clinical Chemistry
Background:
- Accurate quantification of serum methotrexate is crucial for optimizing patient treatment and minimizing toxicity.
- Existing methods for methotrexate measurement may have limitations in terms of speed, sensitivity, or day-to-day variability.
Purpose of the Study:
- To develop and validate a rapid kinetic enzymological assay for determining methotrexate concentrations in human serum.
- To assess the sensitivity, precision, and turnaround time of the novel assay.
Main Methods:
- A kinetic assay utilizing purified dihydrofolic acid reductase from bovine liver was employed.
- Instability of the enzyme was addressed through specific measures, including the use of a reaction rate analyzer.
- Serum samples were analyzed to determine methotrexate levels.
Main Results:
- The assay provides results within 1.5 hours, offering a significant time advantage.
- The lower limit of sensitivity was determined to be 50 nmol/L.
- Day-to-day variation coefficients were approximately 7% in the 200-600 nmol/L range, demonstrating good precision.
Conclusions:
- The described kinetic enzymological assay is a rapid, sensitive, and precise method for serum methotrexate determination.
- This assay offers a viable alternative to existing procedures, potentially improving therapeutic drug monitoring for methotrexate.