Abstract
Publisher Summary Preparation of tetrahydrofolic acid by catalytic hydrogenation of folic acid and by enzymatic reduction of dihydrofolic acid is described in the chapter. A new method of isolation and purification of tetrahydrofolic acid from the crude reaction mixture is also described. The method is based on the observation that tetrahydrofolic acid can be eluted from a DEAE-cellulose (diethylaminoethyl-C) acetate column with diluted acetic acid in the presence of mercaptoethanol. After evaporation of the effluent, a product is obtained that is characterized as a complex between tetrahydrofolic acid and mercaptoethanol of approximate composition— C 19 H 23 N 7 O 6 HS––CH 2 ––CH 2 ––OH . H 2 O with an estimated molecular weight of 541. This compound is slightly more stable than free tetrahydrofolic acid. The method of chromatography is applicable to the synthetic as well as to the enzymatic preparation of tetrahydrofolic acid. The product of DEAE-cellulose chromatography might be contaminated with protein. The crude incubation mixture is recommended to be passed through a Sephadex G-25 column prior to the purification on DEAE-cellulose. The enzymatic synthesis of tetrahydrofolic acid is also discussed.
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