editorial

The interaction vitamin C1

H. P. C. Hogenkamp

between

vitamin

B12 and

Several years ago, Herbert and Jacob (1) H, pKa = 7.8) E#{176} decreases by 60 mV per suggested that the ingestion of megadoses of pH unit. Indeed, at pH 12 the reduction of L-ascorbic acid may produce a vitamin B12 hydroxycobalamin occurs at a potential more deficiency by destroying the cobalamins dur- negative than that of the cob(II)alamin/ ing transport through the gastrointestinal cob(I)alamin couple (7, 8). These redox proptract and possibly during transport and stor- erties are probably the basis for the observations of Herbert et at. (2) that the destructive age in the tissues. More recently, Herbert et at. (2) demonstrated that ascorbate may cause effect of ascorbate is not present, when asspuriously low cobalamin levels in the serum sayed by a radioisotope dilution technique if the vitamin B12 assays are performed with- performed at pH 9.3 (even when KCN is not out the addition of sufficient cyanide. In con- added). The half wave potentials of adenotrast, Marcus et al. (3) concluded that the sylcobalamin in and methylcobalamin (-1.37 V vivo destruction of vitamin B12 by ascorbate and -1.39 V versus SCE) are more negative is highly unlikely because vitamin B12 in food than that of cyanocobalamin (-1.14 V versus and serum was completely unaffected by SCE) indicating that they are not readily ascorbate at room temperature. Furthermore, reduced by ascorbic acid. The polarograms Thenen (4) recently demonstrated that exces- of these three cobalamins show only one two sive intake of L-aSCOrbic acid does not sigelectron wave because at these high potentials nificantly affect the vitamin B12 levels in thethe cobalt atom is reduced to Co (8). The reduction of these cobalamins leads to the plasma and the liver of the rat. in the coordination $ posiThe principal cobalamins in biological ma- loss of the ligand terials are adenosylcobalamin, methylcobaltion. amm, aquocobalamin and its conjugate base, Like cyanocobalamin, adenosytcobalamin, hydroxycobalamin. Cyanocobalamin (viand methylcobalamin are stable at elevated heating aqueous solutions of tamin B12) can be detected in. only some of temperatures; both cobalamins at 94 C for 5 hr did not the biological samples (5). Frost et at. (6) showed in 1952, before the affect their 13C nmr spectra (9). coenzyme forms were discovered, that aquoOne of the most striking properties of adencobalamin is rapidly destroyed by ascorbic osylcobalamin and methylcobalamin is their acid, whereas cyanocobatamin is relatively instability to light. The photolytic decompostable under the same conditions. Aquoco- sition of these two coenzymes involves the homolysis of the carbon-cobalt bond to balamin is readily reduced to cob(II)alamin and organic radicals; in the even by the mildest reducing agents. At neu-cob(II)alamin tral pH the two redox potentials of the aquo- absence of oxygen cob(II)alamin is stable; in cobalamin/cob(II)alamin and the cob(II)the presence of air it slowly oxidizes to aquoalamin/cob(I)alamin couples are -0.04 V and cobalamin. Neither the reduction of aquoco-0.85 V versus SCE, respectively. Above pH 7.8 where hydroxycobatamin becomes the From the Department of Biochemistry, University predominant species (H2OCb1 HOCbt + of Minnesota, Minneapolis, Minnesota 55455.
The American Journal of Clinical Nutrition 33: JANUARY 1980, pp. 1-3. Printed in U.S.A.

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HOGENKAMP

determined that adenosylcobalamin is the batamin to cob(II)alamin and cob(I)alamin nor the phototysis of the coenzymes should major cobalamin in liver (55 to 85%) while lead to modification of the cobalamin moiety. the levels of methylcobalamin and aquocoHowever, most reactions of the cobalamins balamin are much lower (3 5to and 13 to respectively) and, consequently, only 13 that proceed with changes in the oxidation 40%, to of the cobalamin store in the liver state of the cobalt atom are accompanied by 40% would be destroyed by ascorbic acid. In conthe formation of biologically inactive byproducts. trast, in certain diseases involving metabolic In the presence of cyanide ion aquoco- errors in cobalamin metabolism, e.g., failure to synthesize either or both of the cobalamin balamin, hydroxycobalamin and adenosylcoenzymes, aquocobalamin may be the precobalamin are converted to cyanocobalamin; dominant cobalamin. In these rare diseases in contrast, methylcobalamin is stable in the the administration of megadoses of L-ascorbic presence of 0.1 M cyanide in the dark. The first step in the series of reactions that acid could be detrimental. lead to the destruction of the corrin ring by Adenosylcobalamin, methylcobalamin, cyanocobalamin are stable at elevated ascorbic acid is probably the reduction of theand cobalt atom to cob(II)alamin. Indeed, Frost temperatures, 95 to 99 C in neutral solutions. In presence of ascorbic acid aquocoet at. (6) reported that upon the addition of the ascorbic acid the color of the solution balamin of is destroyed almost completely when at 65 C for1/2 hr. Under the same aquocobalamin changed immediately from heated conditions, cyanocobalamin is only slightly red to brown, indicative of cob(II)alamin. The subsequent reactions, which are slower, affected (6); similarly adenosylcobalamin and yield yellow intermediates and eventually methylcobalamin should not be destroyed heated with L-ascorbic acid the dark. in lead to the release of cobalt. These reactions when probably involve hydroxylation and oxida- However, in the light both coenzymes are tion of the corrin ring and its substituents. Lphotolyzed to aquocobalamin and thus cyaAscorbic acid is readily oxidized to dehy- nide is needed to convert the latter cobalamin the cyano form. droascorbic acid when exposed to air, espe- into type addition of cyanide to ascially in the presence of traces of copper and A Kiliani in alkaline solution. This autoxidation gen-corbic acid as suggested by Marcus et al. (3) erates hydrogen peroxide, a strong oxidizing is unlikely because the carbonyl group of a agent (11). In addition, ascorbic acid is ablelactone is not susceptible to nucleophilic atto accelerate the oxidation of many com-tack by cyanide. be useful to routinely add iron or pounds by hydrogen peroxide (12). The re- It may action of L-ascorbic acid and hydrogen per-copper salts to the biological materials to be oxide generates an L-ascorbic acid radical (a extracted for vitamin B12 assays. The metal strong reducing agent) and a hydroxyl radical ions would not only catalyze the auto-oxida(a powerful oxidizing agent). Gossauer andtion of ascorbic acid but also the decomposiGr#{252}ning (13) have recently demonstrated tion of hydrogen peroxide, and thus neutralize the destructive action of vitamin C. that treatment of cyanocobalamin with L-ascorbic acid under aerobic conditions causes In conclusion, it is highly unlikely that hydroxylation at C-S and lactone formation megadoses of vitamin C will destroy all the at C-6 and C-7. cobalamins in the serum and the body stores Thus among the naturally occurring cobal- of healthy subjects. However, large doses of C may precipitate a vitamin B,2 deamins only aquocobalamin is readily reduced vitamin in a few patients suffering from one and subsequently destroyed by L-ascorbic ficiency errors in cobalamin metabolism. fl acid. The major cobalamins in biological ma-or more terials (food, plasma, erythrocytes, liver, etc.) are methylcobalamin and adenosylcoReferences balamin and consequently the destruction of 1. V., AND E. JACOBS. Destruction of vitamin all the cobalamins by L-ascorbate at 37 C is HERBERT, B,2 by ascorbic acid. J. Am. Med. Assoc. 230: 241, highly unlikely, even in the absence of cya1974. nide as long as the cobalamins are not ex-2. HERBERT, V., E. JACOB, K.-T. J. WONG AND R. D. posed to light. For instance, Linnell (5) has PFEFFER. Low serum vitamin B,2 levels in patients

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INTERACTION receiving
cerning

BETWEEN studies con- 8.

vi-

VITAMIN

B,2

AND

3.

4.

5.

6.

7.

LEXA, D., AND J. M. SAVEANT. Thermodynamics and kinetics of oxido-reduction. Third Eurotamin B,2 assay. Am. J. Clin. Nutr. 31: 253, 1978. pean Symposium on Vitamin B,2 and Intrinsic Factor Abstracts 39, 1979. MARCUS, M., M. PRACHUDESAI AND S. WASSEF. H. P. C., P. J. VERGAMINI, AND N. A. Stability of vitamin B,2 in the presence of ascorbic 9. HOGENKAMP, MATWIYOFF. The effect of temperature and light on acid in food and serum: Restoration by cyanide of the carbon13nuclear magnetic resonance spectra of apparent loss. Am. J. Clin. Nutr. In press. THENEN, S. W. High ascorbic acid intake and vialkylcorrinoids, selectively enriched with carbon-i 3. tamin B,2 status in the rat. Third European Sympo-. J. Chem. Soc. (Dalton): 2628, 1975. sium on Vitamin B,2 and Intrinsic Factor Abstracts 10. HOGENKAMP, H. P. C., AND S. HOLMES. Polarography of cobalamins and cobinamides. Biochemistry Ill, Zurich, 1979. 9: 1886, 1970. LINNELL, J. C. The fate of cobalamins in vivo. In: Cobalamins. Biochemistry and Pathophysiology, ed11. WEISSBERGER, A., AND J. E. LUVALLE. Oxidation ited by B. M. Babior. New York: John Wiley & processes XVII. The automation of ascorbic acid in the presence of copper. J. Am. Chem. Soc. 700, 66: Sons, 1975, pp. 287-333. 1944. FROST, D. V., M. LAPIDUS, K. A. PLANT, E. SCHERH. S., H. L. FRUSH. OXIDATION OF L-ascorbic FLING, AND H. H. FRICKE. Differential stability of 12. ISBELL, various analogs of cobalamin to vitamin C. Science acid by hydrogen peroxide: preparation of L-threonic 116: 119, 1952. acid. Carbohydrate Res. 72: 301, 1979. LEXA, D., J. M. SAVEANT, AND J. ZICKLER. Electro13. GOSSAUER, A., AND B. GRUNING. Structure and reactivity of the so-called stable yellow corrinoids. chemistry of vitamin B,2. II. Redox and acid-base Third European Symposium on Vitamin and B,2 equilibria in the B,2./B,2 system. J. Am. Chem. Soc. 99: 2786, 1977. Intrinsic Factor Abstracts 33, Zurich, 1979.

ascorbic

acid
of

in

megadoses:

the

effect

ascorbate

on

radioisotope

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