Hydrochloric Acid → Tetrahydrofolic acid

Participants

Reaction Steps

In the second step, the recovered solid is suspended in water (100 mL) and concentrated ammonium hydroxide (7.2 mL) is added to dissolve the solid and attain a pH of 8.6. The pH of this solution is adjusted to 7.5 by the addition of 6N hydrochloric acid (9 mL). To this solution is added glucose dehydrogenase (20 mg), dihydrofolate reductase (7 mg), NADP (280 mg), glucose (6 g) and water (40 mL). This solution is allowed to stir under N2 for 1 hour. After this time glucose dehydrogenase (16 mg), dihydrofolate reductase (8 mg) and water (20 mL) is added. This solution is allowed to stir for 3.5 hours. The pH of this solution is maintained at 7.5 by the intermittent addition of concentrated ammonium hydroxide. Concentrated hydrochloric acid (9.5 mL) is added to the solution in order to attain a pH of 2.9. The precipitate formed is isolated by centrifugation or other suitable means. The solid is suspended in water (100 mL) and isolated by vacuum filtration. The solid is dried in vacuo at 40° C. overnight to give 5.037 grams of crude tetrahydrofolic acid.

Provenance & Evidence