一种简便、快速测定发酵液中V_(B_(12))含量的方法

A Simple and Rapid Method to Determine Vitamin B_(12) in Fermentation Broth

文中探索了一种不经氰化物转化而直接使用HPLC测定发酵液中维生素B12 (包括羟基钴胺素、脱氧腺苷钴胺素和甲基钴胺素 )的检测方法。将发酵液离心取菌体 ,样品预处理后进样分析 ,采用乙腈和醋酸钠缓冲液为流动相。此方法能够精确检测上述 3种钴胺素 ,并且重复性好、分析速度快 ,可应用于工业发酵生产维生素B12 的过程监控和检测。

The traditional method to determine cyanocobalamin by HPLC cannot be used for rapid determination of vitamin B 12 in fermentation broth. An improved method which measures the contents of hydroxycobalamin, deoxyadenosylcobalamin, methylcobalamin by HPLC so as to directly determine V B 12 is presented. The improved method does not need cyanide transformationm, and makes fast V B 12 detection in industrial process possible. In this paper, V B 12 is produced by Propionibacterium freudenreichii fermentation and cells were collected by centrifugation. After pretreatment, sample is analyzed by HPLC. Acetonitrile and sodium acetate buffer are used as mobile phase in the assay. The results showed that sample can be well separated and the data met the needs of methodology. This method is simple, rapid and accurate, and it can be used in the fermentation process of vitamin B 12 .