High-performance liquid chromatographic measurement of 5,10-methylenetetrahydrofolate in liver.
The folate coenzyme 5,10-methylenetetrahydrofolate is an important folate metabolite which cannot be determined directly by HPLC near neutral pH because it dissociates to formaldehyde and tetrahydrofolate. A method for the determination of 5,10-methylenetetrahydrofolate in liver is described. This method involves (1) determination of liver 5-methyltetrahydrofolate; (2) chemical reduction of liver 5,10-methylenetetrahydrofolate (stabilized at pH 10) to 5-methyltetrahydrofolate; and (3) determination of total liver 5-methyltetrahydrofolate. Subtraction of (1) from (3) gives the concentration of 5,10-methylenetetrahydrofolate in liver.
Horne
D W
DW
Department of Veterans Affairs Medical Center, Nashville, Tennessee 37212, USA. donald.w.horne@vanderbilt.edu
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008099:Liver / Q000737:chemistry*; D008297:Male; D051379:Mice; D010084:Oxidation-Reduction; D051381:Rats; D017207:Rats, Sprague-Dawley; D013763:Tetrahydrofolates / Q000032:analysis*
D013763:Tetrahydrofolates; C013123:5,10-methylenetetrahydrofolic acid; C005984:5-methyltetrahydrofolate