胶束溶液中钙黄绿素-荧光桃红间荧光共振能量转移的研究及应用

Fluorescence Resonance Energy Transfer Between Calcein and Phloxine in Micellar Media

研究了钙黄绿素-荧光桃红体系的荧光共振能量转移机理及其在蛋白质测定中的应用。实验表明:在聚乙烯醇存在下,于HAc-NaAc缓冲液(pH 5.30)中,钙黄绿素与荧光桃红之间能发生有效的能量转移。根据F rster理论,探讨了钙黄绿素与荧光桃红分子间能量转移机理。结果表明:蛋白质的加入使钙黄绿素-荧光桃红体系发生荧光猝灭,而且蛋白质的浓度与体系的荧光猝灭强度在一定范围内有良好的线性关系,据此建立了钙黄绿素-荧光桃红-蛋白质体系测定蛋白质的新方法。在优化条件下,蛋白质的线性范围为0.4～84 mg/L;检出限为0.2 mg/L(n=11)。方法应用于人血清样品中蛋白质含量的测定,其RSD≤1.8%(n=6);回收率为96.9%～102.1%。

Fluorescence resonance energy transfer（FRET） from calcein to phloxine and the application for determining protein was studied.FRET of calcein-phloxine occurred most effectively in aqueous polyvinyl alcohol micellar solutions with HAc-NaAc buffer solution（pH=5.30）.The mechanism of the FRET between calcein and phloxine was investigated.The results showed that the fluorescence of calcein-phloxine system was quenched by adding protein,and there were good linear correlations between the concentrations of protein and the fluorescence quenched intensities.Then a new determination strategy of protein was built by using the calcein-phloxine system.Under optimum conditions,the linear range of determining bovine serum albumin was 0.4-84 mg/L,with a detected limit of 0.2 mg/L（n=11）.The method could be applied to determine the total protein content for the samples of human serum,with RSD〈1.8%（n=6） and the recovery of standard addition was 96.9%-102.1%.