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

荧光标记-HPLC技术高灵敏度糖分析方法的研制

• 批准号: 59880010
• 负责人: IKENAKA Tokuji
• 金额: $ 5.31万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research
• 财政年份: 1984
• 资助国家: 日本
• 起止时间: 1984 至 1985
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-59880010/
• 关键词: HPLC, Sugar analysis; 糖組成分析; 螢光分析; 微量糖分析法

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).

糖蛋白组分糖的高灵敏度分析已经发展起来。该方法的过程是用2 M HCl-2 M三氟乙酸（100 ℃，6 h）水解样品，N-乙酰化，糖的吡啶胺化，使用TSK-GEL G2000 PW柱通过HPLC进行分离，并使用UltraspereODS柱通过HPLC进行糖的吡啶氨基衍生物的色谱测定。该方法允许测定0.01-10 nmol的糖。可分析约0.1 nmol样品，获得满意结果。通过灼烧水解用试管和使用新鲜蒸馏水可降低色谱图上污染葡萄糖峰。通过使用自动进样器（沃茨710 B）和级分收集器（FRAC-100. Pharmacia）。

项目成果

Anal.Biochem.145-1. (1985)

分析.生物化学.145-1。