[ 2 H 4 ]Folic acid was synthesized by deuterating p -aminobenzoic acid, which was then coupled to glutamic acid and 6-formylpterin. Using [ 2 H 4 ]folic acid as starting component enabled the preparation of labeled vitamers tetrahydrofolate, 5-formyltetrahydrofolate, 5-methyltetrahydrofolate, and 10-formylfolate which were characterized by electrospray mass spectrometry and collision-induced dissociation. The mass spectrometric studies confirmed that the compounds could be used as internal standards in stable isotope dilution assays.
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Freisleben et al. (2002) studied this question.
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