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

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).

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