title: Chemical synthesis of deuterated folate monoglutamate and in vivo assessment of urinary excretion of deuterated folates in man.
abstract: the synthesis and in vivo application of stable-isotopically labeled folic acid was investigated to devise methods suitable for studies of folate metabolism in human subjects. Glutamate-labeled tetradeutero-pteroylglutamic acid (d4-folic acid) was prepared by mixed anhydride coupling of N10-trifluoroacetylpteroic acid and dimethyl L-[3,3,4,4-2H4]glutamic acid, saponification in sodium deuteroxide, and chromatographic purification. Retention of the isotopic label was verified by proton NMR and mass spectrometry of the para-aminobenzoylglutamic acid product of C9-N10 bond cleavage. A method was devised for determination of of isotopic enrichment of urinary d4-folates derived from orally administered d4-folic acid using affinity chromatographic purification, chemical cleavage of the C9-N10 bond, HPLC isolation of the p-[2H4]aminobenzoylglutamate product, followed by negative-ion chemical-ionization gas chromatography/mass spectrometry. Data concerning the urinary excretion of d4-folates derived from an oral dose of d4-folic acid in an adult human are presented.
authors.affiliation: Food Science and Human Nutrition Department, University of Florida, Gainesville 32611.
authors.forename: J F
authors.initials: JF
authors.lastname: Gregory
authors.forename: J P
authors.initials: JP
authors.lastname: Toth
mesh_terms: D000284:Administration, Oral; D000328:Adult; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D003903:Deuterium; D005492:Folic Acid / Q000652:urine*; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D009682:Magnetic Resonance Spectroscopy; D008297:Male
chemical_list: D005492:Folic Acid; D003903:Deuterium