Gimsing P, Nexø E, Hippe E
Anal Biochem. 1983 Mar;129(2):296-304. doi: 10.1016/0003-2697(83)90553-5.
An improved method for determination of cobalamins in biological materials is described. The cobalamins are extracted with ethanol after preincubation with cadmium acetate, which inhibits nonspecific adsorption of hydroxocobalamin to proteins. The extracts are desalted on Amberlite XAD-2 columns, which at least doubles the capacity of the analysis, as compared to the previous phenol procedure. The cobalamins are separated by one-dimensional thin-layer chromatography. Bioautography with Escherichia coli strain 113-3 is performed after the light-sensitive cobalamins and hydroxocobalamin have been converted to sulfitocobalamin. This is done in order to ensure comparable growth responses to equal amounts of the cobalamins. Reference intervals of plasma and erythrocytes of healthy blood donors for methyl-, 5'-deoxyadenosyl-, cyano-, hydroxo/aquo-, and sulfitocobalamin are presented. The differences between the present and previous results are discussed.
本文描述了一种用于测定生物材料中钴胺素的改进方法。在与醋酸镉预孵育后，用乙醇提取钴胺素，醋酸镉可抑制羟钴胺素对蛋白质的非特异性吸附。提取物在Amberlite XAD - 2柱上脱盐，与之前的苯酚法相比，这至少使分析能力提高了一倍。通过一维薄层色谱法分离钴胺素。在将光敏钴胺素和羟钴胺素转化为亚硫酸钴胺素后，用大肠杆菌菌株113 - 3进行生物自显影。这样做是为了确保对等量钴胺素具有可比的生长反应。给出了健康献血者血浆和红细胞中甲基钴胺素、5'-脱氧腺苷钴胺素、氰钴胺素、羟钴胺素/水合钴胺素和亚硫酸钴胺素的参考区间。讨论了当前结果与之前结果之间的差异。
