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

点击翻译按钮获取中文摘要

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).