A rapid and specific extraction procedure for folates determination in rat liver and analysis by high-performance liquid chromatography with fluorometric detection.
The native fluorescence of the main biological derivatives of folic acid is maximal at acidic pH. The intensities obtained with 5-methyltetrahydrofolic acid, tetrahydrofolic acid, and formyltetrahydrofolic acid are in the ratio 10/5/1. After separating the metabolites by reversed phase high-performance liquid chromatography, the fluorometric detection limit varied between 0.4 and 20 pmol per injection. Rat liver extract (about 2 g/50 ml of 1% ascorbate solution) could be analyzed after incubation at 37 degrees C overnight, deproteinizing with acetone, and purification and concentration in an ion exchange column. The concentrations obtained varied from 1.27 to 7.96 nmol/g depending on the derivative. Recovery varied from 89 to 113%.
Gounelle
J C
JC
Faculté de Pharmacie, University Paris XI, France.
Ladjimi
H
H
Prognon
P
P
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005470:Fluorometry; D005492:Folic Acid / Q000032:analysis*; D005575:Formyltetrahydrofolates / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000032:analysis*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D013763:Tetrahydrofolates / Q000032:analysis
D005575:Formyltetrahydrofolates; D013763:Tetrahydrofolates; D005492:Folic Acid