The absorption and metabolism of synthetic polyglutamates of folic acid have been compared with free pteroylglutamic acid in four subjects having chronic lymphatic leukemia and one with Hodgkin's granuloma. Pteroylpolyglutamates containing either three or seven glutamate residues were prepared by the solid-phase method permitting placement of carbon-14 labels in either the pteridine ring or in a selected glutamate unit of the gamma peptide chain. Complete dissociation was observed between biological folate activity and radioactivity of plasma after ingestion of pteroyltriglutamate labeled in the middle glutamate. This indicates cleavage to the monoglutamate form at the time of absorption from the intestine or very soon thereafter. A large portion of radioactivity liberated from the middle glutamate is recoverable as carbon dioxide in the exhaled air.

Fecal losses of folate tended to be greater with increasing length of the poly-γ-glutamyl chain. Higher blood levels and greater urinary losses of folate tended to occur after ingestion of mono- and triglutamates than with the heptaglutamate. Calculations based on radioactivity determinations in feces plus urinary folate losses, judged by either radioactivity or microbiological assays, indicated net retention of 37-67% of the dose irrespective of chain length ingested and major avenue of loss.

During the peak of absorption the folate circulating in plasma was active for both Streptococcus fecalis and Lactobacillus casei and carried specific radioactivity which was virtually identical with that of the administered dose. This suggests that neither methylation, conjugation, nor displacement of nonradioactive folate occurred to any significant extent during the 1st 2 hr. The specific radioactivity of 24-hr urine specimens as measured with L. casei corresponded closely with that of the administered dose. Evidence exists that methylation of the radioactive folate may occur, but significant displacement of nonradioactive methylfolate was not observed under the conditions of this study. Since 50-75% of administered heptaglutamate appears to be absorbable in man, estimates of dietary intake should include this fraction as well as the “free” folate.