薄层色谱-薄层扫描法测定发酵液中L-亮氨酸的研究

Study on determining of L-Leucine in fermented liquid by thin-layer chromatography -thin-layer scanning method

用层析硅胶G板进行薄层色谱分离发酵液中L-亮氨酸与L-缬氨酸,经茚三酮显色得到完整的L-亮氨酸色斑;用CS-9301薄层扫描仪测定L-亮氨酸色斑峰面积;根据公式C=(Y-97.286)×n/(2559.5×V)计算发酵液L-亮氨酸浓度。结果表明:该方法平均回收率为97.3%,重现性试验的变异系数为0.045%,说明本方法的准确性和重现性良好,能满足大批量测定发酵液中L-亮氨酸含量的要求。

The combination of thin-layer chromatography and thin-layer scanning may set up a new method to determine the concentrations of L-leucine in fermented liquid in an easy, quick and correct way. The L-leucine and L-valine in fermented liquid can be separated by thin-layer chromatography. After colored by ninhydrin the whole mottles of L-leucine on silica gel-G-board present. Then Using the thin-layer scanner （Shimadzu CS-9301） to determine the chromatographic peak areas of L-leucine mottles, the concentrations of L-leucine were calculated according to the following formula, C=（Y-97.286）xn/（2559.5xV）. It is indicated that the veracity and recurrence of this method are quite well as its rate of reclaiming is 97.3% and the RSD is 0.045%, it can be used to determine the concentrations of L-leucine in fermented liquid on a large-scale.