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  

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

作　　者：蔡沅君 关升 晏国全 张祥民[1,2] CAI Yuan-Jun;GUAN Sheng;YAN Guo-Quan;ZHANG Xiang-Min(Department of Chemistry,Fudan University,Shanghai 200433,China;Institute of Biomedical Sciences,Fudan University,Shanghai 200030,China)

机构地区：[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

出　　处：《分析化学》2021年第9期1540-1545,共6页Chinese Journal of Analytical Chemistry

基　　金：国家重点研发计划项目(No.2017YFA0505003);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。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(R^(2)=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.

关 键 词：糖化白蛋白 反相色谱 唾液 葡萄糖苯脎法 无创检测 蛋白定量分析 

分 类 号：R446.1[医药卫生—诊断学] O657.7[医药卫生—临床医学]