Measurements of sub-nanomolar concentrations of unmetabolised folic acid in serum.

We describe a combined HPLC/microbiological assay procedure for the sub-nanomolar analysis of unmetabolised folic acid (pteroylglutamate) in human serum. This metabolically unaltered form of the vitamin arises following the consumption of folic acid either in supplemental form or in fortified foods. Following HPLC separation of folic acid from other folate derivatives the folic acid fraction was concentrated by C(18) Sep-Pak cartridges and assayed by Lactobacillus casei microbiological assay.