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Journal of Pharmacology And Experimental Therapeutics, Vol. 137, Issue 2, 162-166, 1962
Copyright © 1962 by American Society for Pharmacology and Experimental Therapeutics

ASSAY FOR 4-AMINO FOLIC ACID ANALOGUES BY INHIBITION OF FOLIC ACID REDUCTASE

William C. Werkheiser 1, Sigmund F. Zakrzewski 1, and Charles A. Nichol 1

1 Department of Experimental Therapeutics, Roswell Park Memorial Institute, Buffalo, New York

A precise, specific assay for the 4-amino analogues of folic acid in urine, serum and tissues has been described. The method is based on the stoichiometric inhibition of the enzyme folic acid reductase by these drugs. A titration of drug with enzyme is measured by a colorimetric determination (Bratton-Marshall reaction) of the diazotizable p-aminobenzoylglutamate

derived from the enzymic product, tetrahydrofolate. This method is specific for the unaltered form of these drugs and is free from interference by the presence of growth factors such as folic acid, folinic acid or thymine. The quantitative determination of folic acid by the measurement of diazotizable amine following complete reduction of the substrate by the enzyme has also been demonstrated.

Submitted on March 15, 1962




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Copyright © 1962 by the American Society for Pharmacology and Experimental Therapeutics.