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Microdetermination of proteins based on enhancement effect of Rayleigh light scattering of hematoxylin in hot water bath

Baohong Chong, Baosheng Liu, Zhiyun Li, Shina Cao, Xiaona Yan


A highly sensitive Rayleigh light scattering (RLS) method has been developed for the determination of protein concentrations in human serum album( HSA) and bovine serumalbum(BSA) based on the reactions of HSA and BSA with oxidized hematoxylin. One of the distinguishing features of this method proposed here was heating the mixture of protein and hematoxylin at pH 4.40 in 80 oC water bath for 15 minutes before assay. And another interesting phenomenon observed was that the addition of hematoxylin to the protein solution could prevent protein fromdeposition in hot water. Under the optimal experimental conditions, the greatly enhanced RLS of dye-protein was linearly. The linear range of the determination is 0- 6.2 mg/L for HAS, and the detection limit is 34 ìg. The foreign substances, such as urea, amino acids and metal ions, have little effect on the determination.


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