三种L-苏氨酸的测定方法比较

Comparison of Three Methods for Determination of L-threonine

目的:比较3种测定L-苏氨酸含量的方法。方法:对三种L-苏氨酸的测定方法:纸层析法、薄层层析法、高效液相PITC柱前衍生法进行了比较和优化。并用以上三种方法测定10份发酵液样品中的L-苏氨酸含量及加标回收率。结果:纸层析法、薄层层析法和高效液相PITC柱前衍生法三种方法的决定系数分别为0.9938、0.9971和0.9992,检出限分别为:5、7和1.2μg/L。在每组样品的方差方面,高效液相PITC柱前衍生法方差最小,纸层析法和薄层层析法相近。三种方法平均样品加标回收率分别为95.60%、95.10%和99.30%。三种方法用来检测样品中的L-苏氨酸含量均效果良好。在准确性方面,高效液相PITC柱前衍生法的准确性最好,纸层析法和薄层层析法次之。结论:纸层析法和薄层层析法成本低,适合部分对精确度要求不高的含量测定。高效液相PITC柱前衍生法测定精确,成本较高,速度快,适合精确测定。

Objective:To compare three methods for the determination of L-threonine.Methods:Three L-threonine determination methods:paper chromatography,thin layer chromatography,high performance liquid phase PITC pre-column derivatization were compared and optimized.And the above three methods were used to determine the L-threonine content and spike recovery in 10 fermentation broth samples.Results:The determination coefficient of paper chromatography,thin layer chromatography and high performance liquid phase PITC pre-column derivatization were 0.9938,0.9971 and 0.9992,and the detection limits were 5,7 and 1.2μg/L.In terms of the variance of each group of samples,the difference of the high performance liquid phase PITC pre-column derivatization was the smallest,and the paper chromatography was similar to the thin-layer chromatography.The average recoveries of the three methods were 95.60%,95.10%,and 99.30%.Conclusion:The three methods are all effective for detecting the content of L-threonine in samples.In terms of accuracy,high performance liquid phase PITC pre-column derivatization has the best accuracy,followed by paper chromatography and thin layer chromatography.In summary,paper chromatography and thin-layer chromatography are low-cost and suitable for content determinations that do not require high accuracy.High performance liquid phase PITC pre-column derivatization is accurate,high cost,fast and suitable for accurate determination.