Oxidative and reductive cleavage of folates--a critical appraisal.

Abstract A critical analysis has been made of the oxidative and reductive techniques employedfor cleavage of the C 9 -N 10 bond of folic acid and its derivaatives. The assumption has previously been made that these cleavage reactions reduce folates to a common family of p -aminobenzoylglutamate derivatives varying only in the lengths of γ -polyglutamyl peptide side chains which are readily subjected to quantitative and qualitative analysis. This assumption is incorrect. Oxidation by potassium permanganate effectively cleaved folic acid, dihydrofolic acid, tetrahydrofolic acid, and 5-formyltetrahydrofolic acid to yield p -aminobenzoylglutamate. 5-Methyltetrahydrofolic acid was merely oxidized to 5-methyldihydrofolic acid while 5,10-methenyltetrahydrofolic acid and 10-formyltetrahydrofolic acid were oxidized to 10-formylfolate which was stable to further attack. Of all the folate derivatives tested only folic acid and dihydrofolic acid were cleaved to p -aminobenzoylglutamate by the zinc-hydrochloric acid reduction method. Both tetrahydrofolic acid and 5-methyltetrahydrofolic acid were stable under fully reducing conditions. 5,10-Methenyl-,10-formyl-, and 5-formyltetrahydrofolic acid yielded N -methyl- p -aminobenzoylglutamate. It is evident, therefore, that not only is the dominant mammalian tissue folate derivative, 5-methyltetrahydrofolate, resistant to cleavage by either method, but that a common family of p -aminobenzoylglutamate derivatives is not the end product of those folate compounds that are susceptible. While this may not invalidate the reports of the relative polyglutamate chain lengths of tissue folates such data should be regarded with some caution.