Folic acid purification

Xanthopterin monohydrate (2-amino-4,6-dihydroxypteridine, 2-amino-pteridin-4,6(lff,5ff)-dione) [5979-01-1 (H2O), 119-48-8 (anhydr)] M 197.2, m <300", pK, 1.6 (basic), pKj 6.59 (acidic), PK3 9.31 (acidic)(anhydrous species), and pKj 1.6 (basic), pK2 8.65 (acidic), PK3 9.99 (acidic)(7,8-hydrated species). Purification as for isoxanthopterin. Crystd by acidifying an ammoniacal soln, and collecting by centrifugation followed by washing with EtOH, ether and drying at 100° in vacuo. Paper chromatography Rp 0.15 ( -PrOH, 1% aq NH3, 2 1), 0.36 ( -BuOH,AcOH, H2O, 4 1 1) and 0.47 (3% aq NH3). [Inoue and Perrin J Chem Soc 260 7962 Inoue Tetrahedron 20 243 I964 see also Blakley Biochemistry of Folic Acid and Related Pteridines North Holland Publ Co, Amsterdam 1969.]... 

Chapter 10 is devoted to the preparation and purification of hydrophilic vitamins (C, Bj, Bj, Bg, B[2, nicotinic acid and nicotinamide, pantothenic acid, biotin, and folic acid) in pharmaceutical preparations, food products, and biological samples. 

Walter, R. D., and Konigk, E. (1974a). Biosynthesis of folic acid compounds in plasmodia. Purification and properties of the 7,8-dihydropteroate-synthesizing enzyme from Plasmodium chabaudi. Hoppe Seylers. Z. Physiol. Chem. 355,431-437. 

Claradiastase and trypsin in phosphate buffer have been used for hydrolysis of sample matrix for the determination of supplemental folic acid (27). Jacoby and Henry (26) further modified the method of Hoppner and Lampi (41) for folic acid by HPLC, in which the folic acid added to infant formulas and liquid medical nutritionals is quantitatively extracted with the aid of bacterial protease and papain. One disadvantage of the enzymatic extraction method was the large number of UV-absorbing compounds that are formed during enzymatic hydrolysis. These can interfere in the quantitation of the folate peaks. Thus, addition of a-amylase and protease to enhance extraction has not been common practice for methods involving HPLC. However, Pfeiffer et al. (13) suceessfiilly used triple-enzyme treatment prior to HPLC analysis with a purification method based on affinity chromatography. 

Folic acid (17) Purification of low-mol.-wt. human folate-binding protein by affinity chromatography 101... 

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