Stupperich E, Steiner I, Rühlemann M
Anal Biochem. 1986 Jun;155(2):365-70. doi: 10.1016/0003-2697(86)90447-1.
Cyanocobamides were extracted from diverse bacterial species, purified by XAD-4 and neutral aluminum oxide column chromatography, and separated by isocratic reversed-phase high-performance liquid chromatography (HPLC). Retention times are given for seven cobamide types: dicyanocobinamide (factor B), Co alpha-(alpha-benzimidazolyl)-Co beta-cyanocobamide, Co alpha-(5-hydroxybenzimidazolyl)-Co beta-cyanocobamide (factor III), Co alpha-(5-methoxybenzimidazolyl)-Co beta-cyanocobamide (factor IIIm), Co alpha-(5-methylbenzimidazolyl)-Co beta-cyanocobamide, cyanocob(III)alamin (vitamin B-12) and Co alpha-(naphthimidazolyl)-Co beta-cyanocobamide. Other Co beta-ligandyl-cobamides such as hydroxycobamide and the light-sensitive methyl-, acetyl-, propyl-, and adenosylcobamides were separated by HPLC in a gradient mode. The recovery of total cell cobamide after extraction, purification, and separation was 75-80%. The method was useful in preparative and analytical work. Less than 10 ng cyanocobamide was detectable.
从不同细菌物种中提取氰钴胺，通过XAD - 4和中性氧化铝柱色谱法进行纯化，并通过等度反相高效液相色谱法（HPLC）进行分离。给出了七种钴胺素类型的保留时间：二氰钴胺素（因子B）、Coα -（α - 苯并咪唑基）- Coβ - 氰钴胺、Coα -（5 - 羟基苯并咪唑基）- Coβ - 氰钴胺（因子III）、Coα -（5 - 甲氧基苯并咪唑基）- Coβ - 氰钴胺（因子IIIm）、Coα -（5 - 甲基苯并咪唑基）- Coβ - 氰钴胺、氰钴胺（III）（维生素B - 12）和Coα -（萘并咪唑基）- Coβ - 氰钴胺。其他Coβ - 配体钴胺素，如羟基钴胺以及对光敏感的甲基钴胺、乙酰钴胺、丙基钴胺和腺苷钴胺，通过HPLC以梯度模式进行分离。提取、纯化和分离后细胞总钴胺素的回收率为75 - 80%。该方法在制备和分析工作中很有用。可检测到的氰钴胺含量低于10 ng。
