反相色谱分离和葡萄糖苯脎法测定人唾液中白蛋白的糖化水平

引用本文: 蔡沅君, 关升, 晏国全, 张祥民. 反相色谱分离和葡萄糖苯脎法测定人唾液中白蛋白的糖化水平[J]. 分析化学, 2021, 49(9): 1540-1545. doi: 10.19756/j.issn.0253-3820.211200 shu

Citation: CAI Yuan-Jun, GUAN Sheng, YAN Guo-Quan, ZHANG Xiang-Min. Chromatographic Separation and Quantitative Analysis of Glycated Albumin in Human Saliva by Glucose Phenylosazone Spectrophotometric Assay[J]. Chinese Journal of Analytical Chemistry, 2021, 49(9): 1540-1545. doi: 10.19756/j.issn.0253-3820.211200 shu

反相色谱分离和葡萄糖苯脎法测定人唾液中白蛋白的糖化水平

蔡沅君 ^1, ,
关升 ^2, ,
晏国全 ^{2,
,} ,
张祥民 ^{1,2,
,}

1.
复旦大学化学系, 上海 200433;
2.
复旦大学生物医学研究院, 上海 200030

通讯作者: 晏国全,E-mail:yanguoquan@fudan.edu.cn; 张祥民,E-mail:xmzhang@fudan.edu.cn

基金项目:
国家重点研发计划项目（No.2017YFA0505003）和国家自然科学基金项目（No.21775027）资助。

摘要: 采用一种无创取样方法实现了人体内糖化白蛋白的定量分析。以唾液为测试样品，首先通过超滤分离出唾液中分子量大于10 kD的蛋白分子，再经过反相色谱对唾液中的白蛋白进行进一步分离提纯，其中36.0~37.0 min的馏分经质谱鉴定为人血清白蛋白。通过葡萄糖苯脎分光光度法对唾液馏分中的白蛋白进行定量分析，并与临床中使用的白蛋白定量分析方法进行比较。结果表明，对于健康样品，5次平行实验的平均糖化比例为10.9%，相对标准偏差为11.3%。本方法可定量分析健康人唾液中的糖化白蛋白，有望用于临床糖尿病初步筛查。

关键词:

English

Chromatographic Separation and Quantitative Analysis of Glycated Albumin in Human Saliva by Glucose Phenylosazone Spectrophotometric Assay

1.
Department of Chemistry, Fudan University, Shanghai 200433, China;
2.
Institute of Biomedical Sciences, Fudan University, Shanghai 200030, China

Corresponding author: YAN Guo-Quan, ; ZHANG Xiang-Min,

Received Date: 16 March 2021
Revised Date: 30 May 2021
Fund Project:
Supported by the National Key Research and Development Program of China (No.2017YFA0505003) and the National Natural Science Foundation of China (No.21775027).

Abstract: Diabetes mellitus, characterized by hyperglycemia, is a worldwide disease which may lead to chronic complications. In the process of diabetes detection and monitoring, blood sampling is essential. In this work, a non-invasive, blood-free method was developed for the quantification of glycated albumin in saliva. The saliva sample first underwent size seizing to obtain proteins with molecular weight of larger than 10 kD, then it was separated using the reversed phase liquid chromatography (RPLC), and 36.0-37.0 min fraction was collected. Glycated salivary albumin was quantified by the glucose phenylosazone spectrophotometric assay, which showed good linearity (R²=0.997) in the range 5.0×10^-4-5.0×10^-2 mmol/L. With this method, the glycated albumin value (GA value) and reproducibility (RSD) for five parallel experiments were 10.9% and 11.3%, respectively. In addition, the GA values of four healthy volunteers proved the method feasible. In conclusion, this method was sensitive and reliable for the quantification of GA values in human saliva, and was inspiring for the clinical detection of diabetes mellitus.

Key words:

Glycated albumin
/ Reversed phase liquid chromatography
/ Saliva
/ Glucose phenylosazone spectrophotometric assay
/ Non-invasive detection
/ Protein quantification

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沈阳化工大学材料科学与工程学院沈阳 110142

反相色谱分离和葡萄糖苯脎法测定人唾液中白蛋白的糖化水平

通讯作者: 晏国全,E-mail:yanguoquan@fudan.edu.cn; 张祥民,E-mail:xmzhang@fudan.edu.cn

English

Chromatographic Separation and Quantitative Analysis of Glycated Albumin in Human Saliva by Glucose Phenylosazone Spectrophotometric Assay

Corresponding author: YAN Guo-Quan, ; ZHANG Xiang-Min,

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

反相色谱分离和葡萄糖苯脎法测定人唾液中白蛋白的糖化水平

通讯作者: 晏国全,E-mail:yanguoquan@fudan.edu.cn; 张祥民,E-mail:xmzhang@fudan.edu.cn

English

Chromatographic Separation and Quantitative Analysis of Glycated Albumin in Human Saliva by Glucose Phenylosazone Spectrophotometric Assay

Corresponding author: YAN Guo-Quan, ; ZHANG Xiang-Min,

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

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