Development of highly sensitive analytical method of sugars by fluorescence labeling-HPLC technique

对糖蛋白质成分糖的高度敏感的分析已经发展出来。该方法的过程是用2 M HC 1 -2 M三氟乙酸（100 ° C， 6 h）、N-乙酸化、糖的吡咯烷化、用HPLC对TSK-GELG 2000 PW柱进行降解，并用HPLC对蔗糖的吡咯烷胺衍生物进行色谱测定。该方法允许测定0.01-10纳摩尔的糖。关于0.1 nmol的样品可以用满意的结果进行分析。色谱仪上的葡萄糖峰值可以通过燃烧的测试管来减少，并通过使用新鲜的分散的水来减少。通过使用自动注射器（Waters 710 B）和片段收集器（FRAC-100）实现分析的自动化。Pharmacia）

Highly sensitive analysis of component sugars of glycoproteins has been developed. The processes of the method are hydrolysis of a sample with 2 M HC1-2 M trifluoroacetic acid (100゜C, 6 h), N-acetylation, pyridylamination of sugars, desalting by HPLC using a TSK-GEL G2000PW column, and chromatographic determination of pyridylamino derivatives of sugars by HPLC using a UltrasphereODS column. The method allowed the determination of 0.01-10 nmol of sugars. About 0.1 nmol of a sample can be analyzed with satisfactory results. Contaminating glucose peak on chromatogram could be reduced by burnning test tubes used for hydrolysis and by using freshly distilled water. Automation of the analysis was achieved by using an automatic injector (Waters 710B) and a fraction collector (FRAC-100. Pharmacia).