Abstract：Using 1,6-hexanediaminediacridine as a spectrum probe,a new method of quantitative determination of protein by resonance Rayleigh scattering (RRS) has been developed. Characteristics of the resonance scattering reaction of the acridine probe with bovine serum albumin (BSA) were investigated. Effects of the concentration of acridine probe and pH value of the buffer solution,as well as the stability of the resonance scattering interaction,were also studied,and then the optimum condition was obtained. In the Tris-HCl buffer solution,the intensity of resonance Rayleigh scattering spectrum remained stable without any notable change in the pH range of 6.0-7.5. While pH ranged from 8.0 to 8.9,the interaction of BSA and acridine was accelerated. The resonance scattering was increasing and the peak value was gained at pH 8.9. However,with the augmentation of pH from 10.0 to 11.0 in Na2B4O7-NaOH or Na2HPO4-NaOH buffer medium，some negative effects on the molecular structure and nature of BSA might occur and resulted in a too high background scattering noise,which caused a rapid decrease in the resonance scattering intensity. A good calibration curve of the protein was obtained in the range of 1.00-5.00 μg·mL-1 with a detection limit (3σ/K) of 0.085 μg·mL-1. Applied to the quantitative analysis of BSA simulant samples,the result was satisfied and the recoveries were 98.0%-101.7% at the concentrations ranging from 2.00 to 4.00 μg·mL-1. The relative standard deviation was less than 1.7%.
Key words：1;6-Hexanediaminediacridine;Determination of protein;Resonance scattering
林旭聪,吴孟辉,郭良洽,谢增鸿*. 新的光散射体系测定蛋白质的研究[J]. 光谱学与光谱分析, 2007, 27(08): 1587-1590.
LIN Xu-cong,WU Meng-hui,GUO Liang-qia,XIE Zeng-hong*. Studies on the Determination of Protein by a New Resonance Rayleigh Scattering System. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2007, 27(08): 1587-1590.
