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A Note on the Stability of Vitamin B12 in the Presence of Thiochrome*
A Note on the Stability of Vitamin BIZ in the Presence of Thiochrome* By LOUIS J. RAVIN and ROBERT F. DOERGE
I
review article Scheindlin (1) discusses the chemical interactions of the water-soluble vitamins. H e points out that while several studies have been published (2-6) concerning the stability of vitamin BE in the presence of thiamine and nicotinamide, much remains t o be learned about the reaction between thiamine and nicotinamide, as well as the reaction between thiamine and vitamin BIZ. In addition to the references cited by Scheindlin ( l ) , additional information on this problem has been reported by Gambier and Rahn (7,8). Several of these studies have implicated the thiazole moiety of thiamine in the breakdown of vitamin BIZ. however, no mention has been made concerning the possible role of thiochrome. This note presents some preliminary data regarding the possible interaction of thiochrome, a n oxidation product of thiamine, with vitamin Biz in aqueous buffered solution. N A RECENT
TABLEI.-STABILITYOF CRYSTALLINE VITAMINBu IN AQUEOUSBUFFERED SOLUTION CONTAINING THIOCHROMR4
Storage Condition
Original 85'C. RT 45OC. 60°C. RT 45OC. 60°C. RT 45°C. 6OoC.
Storage Time
Solution Containing Thiochrome. pg./5 ml.
..... 8 hours 16 hours 24 hours 3 weeks 3 weeks 3 weeks 7 weeks 7 weeks 7 weeks 3 months 3 months 3 months
28 28 26 26 29 27 29 29 27 25 27 27 23
Control, ~g.i5 ml.
28 28 27 26 29 30 29 26 24 24
28 26 22
8.5 mg./5 ml.. equivalent to 11 mg./5 ml. thiamine hydrochlonde. (I
XXPERIMENTAL
Thiochrome obtained from Bios Laboratories Inc. and cyanocobalamin U. S. P. (crystalline vitamin BIZ, Merck and Co.) were used in these studies. Solutions containing 25 pg. of crystalline vitamin BIZand 8.5 mg. of thiochrome' in each 5 ml. were prepared and adjusted to pH 4.0. A control solution containing 25 pg. of crystalline vitamin B1z was also prepared. The finished solutions were filtered through a Selas* unglazed porcelain candle, 02 porosity, and packaged into 1-02. amber bottles. Samples were set down at RT. 45, 60, and 85" and assayed periodically for vitamin BIZ by the U. S. P. microbiological method (9). DISCUSSION
The assay data obtained in this study are summarized in Table I.
* Received September 20, 1958, from the Research and Development Division of Smith Kline and French Laboratories, Philadelphia 1 Pa. 1 Equivalent' to 11 mg./5 ml. thiamine hydrochloride. 2 Selas Corporation of America, Philadelphia, Pa.
The above data clearly indicate that thiochrome is not responsible for accelerating the decomposition of vitamin B12 in a n aqueous buffered solution under these experimental conditions. Additional studies concerning the interaction between thiamine and vitamin BIZ which also include the possible reaction with thiamine decomposition products are in progress. A complete report on the results of these studies will be forthcoming. REPBRENCES (1) Scheindlii, S., Drug & Cosmetic Ind., 83, 46(1958). ( 2 ) Blitz, M., Eigen, E., and Gunsberg, E.. THISJOURNAL.
45. 803(1956). (3) Blitz. M., Eigen, E., and Gunsberg. E., ibid., 43, 651 (1954). (4) Mukherjee. S. L., and Sen, S. P., J . Pharm. and Pharmocol. 9 759(1957). (5) Ma&k.'T. J., and Feller, B. A., THISJOURNAL, 44, 254(1955). ( 6 ) Feller, B. A.. and Macek, T. J., ibid., 44, 662(1955). (7) Gambier, A. S . . and Rahn, E. P. G., ibid.. 46, 134
\."".,. fiarrm
(8) Gambier, A. S.. and Rahn, E. P. G., ;bid., 47, 356 (1958). (9)"United States Pharmaeopeia." 15th rev.. Mack Publishing Co., Easton. Pa., 1955, p. 885.
Book Notices Handbook of Toxicology. Vol. IV. Tranquilizers. Edited by RUDOLPH M. GREBE. W. B. Saunders Co.,West Washington Square, Philadelphia, Pa., 1959. Viii 4-120 pp. Paperbound. Price $4. This handbook is the eighth of a continuing series of publications, each containing information, chiefly tabular, in one or more of t h e biological sciences. All of the volumes in the series have been prepared under the general direction of the a m mittee on the Handbook of Biological Data, Division of Biology and Agriculture, National Academy of Sciences National Research Council. This latest volume furnishes, in outline form, pertinent data on
26 tranquilizers which have been used as psychopharmacological agents. In general, the information about each of the drugs in this class includes the nonproprietary name, the trademark, the name of the manufacturer. the mdecular formula and weight, the structure, a brief description of the physical and chemical Properties, the Ph~macolotD', certain clinical data, the toxicity, and the mode and site of action. The book is documented with the more important references t o the Ori&d 1iteratUre. The book is well designed to furnish information in summary form on the principal tranquilizers now being used.
425
I
review article Scheindlin (1) discusses the chemical interactions of the water-soluble vitamins. H e points out that while several studies have been published (2-6) concerning the stability of vitamin BE in the presence of thiamine and nicotinamide, much remains t o be learned about the reaction between thiamine and nicotinamide, as well as the reaction between thiamine and vitamin BIZ. In addition to the references cited by Scheindlin ( l ) , additional information on this problem has been reported by Gambier and Rahn (7,8). Several of these studies have implicated the thiazole moiety of thiamine in the breakdown of vitamin BIZ. however, no mention has been made concerning the possible role of thiochrome. This note presents some preliminary data regarding the possible interaction of thiochrome, a n oxidation product of thiamine, with vitamin Biz in aqueous buffered solution. N A RECENT
TABLEI.-STABILITYOF CRYSTALLINE VITAMINBu IN AQUEOUSBUFFERED SOLUTION CONTAINING THIOCHROMR4
Storage Condition
Original 85'C. RT 45OC. 60°C. RT 45OC. 60°C. RT 45°C. 6OoC.
Storage Time
Solution Containing Thiochrome. pg./5 ml.
..... 8 hours 16 hours 24 hours 3 weeks 3 weeks 3 weeks 7 weeks 7 weeks 7 weeks 3 months 3 months 3 months
28 28 26 26 29 27 29 29 27 25 27 27 23
Control, ~g.i5 ml.
28 28 27 26 29 30 29 26 24 24
28 26 22
8.5 mg./5 ml.. equivalent to 11 mg./5 ml. thiamine hydrochlonde. (I
XXPERIMENTAL
Thiochrome obtained from Bios Laboratories Inc. and cyanocobalamin U. S. P. (crystalline vitamin BIZ, Merck and Co.) were used in these studies. Solutions containing 25 pg. of crystalline vitamin BIZand 8.5 mg. of thiochrome' in each 5 ml. were prepared and adjusted to pH 4.0. A control solution containing 25 pg. of crystalline vitamin B1z was also prepared. The finished solutions were filtered through a Selas* unglazed porcelain candle, 02 porosity, and packaged into 1-02. amber bottles. Samples were set down at RT. 45, 60, and 85" and assayed periodically for vitamin BIZ by the U. S. P. microbiological method (9). DISCUSSION
The assay data obtained in this study are summarized in Table I.
* Received September 20, 1958, from the Research and Development Division of Smith Kline and French Laboratories, Philadelphia 1 Pa. 1 Equivalent' to 11 mg./5 ml. thiamine hydrochloride. 2 Selas Corporation of America, Philadelphia, Pa.
The above data clearly indicate that thiochrome is not responsible for accelerating the decomposition of vitamin B12 in a n aqueous buffered solution under these experimental conditions. Additional studies concerning the interaction between thiamine and vitamin BIZ which also include the possible reaction with thiamine decomposition products are in progress. A complete report on the results of these studies will be forthcoming. REPBRENCES (1) Scheindlii, S., Drug & Cosmetic Ind., 83, 46(1958). ( 2 ) Blitz, M., Eigen, E., and Gunsberg, E.. THISJOURNAL.
45. 803(1956). (3) Blitz. M., Eigen, E., and Gunsberg. E., ibid., 43, 651 (1954). (4) Mukherjee. S. L., and Sen, S. P., J . Pharm. and Pharmocol. 9 759(1957). (5) Ma&k.'T. J., and Feller, B. A., THISJOURNAL, 44, 254(1955). ( 6 ) Feller, B. A.. and Macek, T. J., ibid., 44, 662(1955). (7) Gambier, A. S . . and Rahn, E. P. G., ibid.. 46, 134
\."".,. fiarrm
(8) Gambier, A. S.. and Rahn, E. P. G., ;bid., 47, 356 (1958). (9)"United States Pharmaeopeia." 15th rev.. Mack Publishing Co., Easton. Pa., 1955, p. 885.
Book Notices Handbook of Toxicology. Vol. IV. Tranquilizers. Edited by RUDOLPH M. GREBE. W. B. Saunders Co.,West Washington Square, Philadelphia, Pa., 1959. Viii 4-120 pp. Paperbound. Price $4. This handbook is the eighth of a continuing series of publications, each containing information, chiefly tabular, in one or more of t h e biological sciences. All of the volumes in the series have been prepared under the general direction of the a m mittee on the Handbook of Biological Data, Division of Biology and Agriculture, National Academy of Sciences National Research Council. This latest volume furnishes, in outline form, pertinent data on
26 tranquilizers which have been used as psychopharmacological agents. In general, the information about each of the drugs in this class includes the nonproprietary name, the trademark, the name of the manufacturer. the mdecular formula and weight, the structure, a brief description of the physical and chemical Properties, the Ph~macolotD', certain clinical data, the toxicity, and the mode and site of action. The book is documented with the more important references t o the Ori&d 1iteratUre. The book is well designed to furnish information in summary form on the principal tranquilizers now being used.
425