The Chemistry of Anti-Pernicious Anæmia Factors

Abstract

Our previous conclusion that acid hydrolysis of vitamin B12 leads to the formation of a “ninhydrin-reacting” substance is confirmed. The behaviour of this “ninhydrin-reacting” substance and of 2-aminopropanol on paper chromatograms has been examined. The results appear to show that these two substances are identical. Evidence has been obtained for the presence in acid hydrolysates of vitamin B12 of material showing selective absorption with bands and inflections at 2850, 2768, 2690, 2585 and 2500 å (referred to as “the 285 component“). This “285 component” is resolved into at least two structurally related substances by chromatography employing n-butyl alcohol-acetic acid as the irrigation solvent. Formation of ammonia occurs during acid or alkaline hydrolysis of vitamin B12. The authors thank Mr. R. E. Rodway for technical assistance in the isolation of crystalline vitamin B12 from a concentrate kindly supplied to the Research Department by Mr. A. W. Davidson. The ultra-violet absorption data were kindly determined by Dr. E. R. Holiday (M.R.C. Spectrographic Unit The London Hospital, E.1***), who also carried out the examinations with the Corning filter. The authors thank the Directors of The British Drug Houses, Ltd., for permission to publish these results.