液相色谱法测定生物发酵液中水溶性维生素的研究

Study on Determination of Water soluble Vitamins in the Solution of Biological Fermentation by HPLC

本文采用反相离子对色谱测定了生物发酵液中的七种水溶性维生素。提出了用自行装填的酸性氧化铝（０．１９～０．１５ｍｍ）前置柱，和５％乙酸洗脱样品的前处理方法；并就有关的色谱条件进行了选择。色谱柱为ＮｏｖａＰａｋＣ１８；流动相Ａ液为０．０５ｍｏｌ／Ｌ庚烷磺酸盐（ＰＩＣＢ７），冰乙酸调ｐＨ＝２．５；Ｂ液为甲醇＋三乙胺（１００＋０．５）；梯度洗脱；ＵＶ２５４ｎｍ、２９０ｎｍ、３６０ｎｍ同时测定。线性范围为０．１～１０μｇ，各种维生素的ＲＳＤ在１．２％～３．０％之间（ＶｉｔａｍｉｎＢ１２除外），回收率大于９０％。

A HPLC method for determination of water soluble vitamins in the solution of biological fermentation is reported.The samples were processed by a per column packed with acidic aluminum oxide before injected in chromatographic column.A NovaPak C 18 as the chromatographic column,0.05 mol/L PIC B7 solution adjusted pH 2.5 by glacial acetic acid (A) and methanol+triethylamine(100+0.5)(B) as mobile phase for gradient elution were adopted in HPLC analysis.Seven kinds of vitamins are detected at UV 254 nm,290 nm and 360 nm with a linear range of 0.1～10 μg,RSD of 1.2%～3.0%(except Vitamin B 12 ).The recoveries all are more than 90%.