吖啶橙-罗丹明6G荧光共振能量转移及其罗丹明6G荧光猝灭法测定蛋白质

研究了吖啶橙(AO)与罗丹明6G(R6G)之间能量转移的最佳条件。在pH=7．20的Tirs-HCl缓冲溶液，十二烷基苯磺酸钠介质中，AO-R6G能够发生有效能量转移，使R6G荧光增强。蛋白质的加入使R6G荧光猝灭，以此建立了利用AO-R6G荧光共振能量转移间接测定蛋白质的新方法。牛血清白蛋白、人血清白蛋白工作曲线线性范围分别为1．0～31和1．0—30mg／L；检出限分别为0．32和0．33mg／L；平行6次测定相对标准偏差为1．1％～2．0％；回收率为96．7％～103．2％。此方法的稳定性好，选择性高，用于人血清试样中总蛋白含量的测定，与常用的双缩脲法基本一致。

Fluorescence Resonance Energy Transfer between Acridine Orange and Rhodamine 6G and Fluorescence Quenching of Rhodamine 6G Reaction for the Determination of Proteins

An optimum experimental conditions of energy transfer between acridine orange (AO) and rhodamine 6G (R6G) were studied. It was found that the effective energy transfer could occur between AO and R6G in the sodium dodecyl benzene sulfonate solution containing Tris-HCl buffer solution (pH=7.20), which improved the fluorescence intensity of R6G . The fluorescence of AO-R6G systems was quenched with the addition of proteins in alkalescence medium. Therefore, a novel fluorescence quenching method of indirect determination for proteins was built with the AO-R6G fluorescence resonance energy transferred. The linearity of calibration curves for the determination of bovine serum albumin (BSA) and human serum albumin (HSA) was 1.0~31 mg/L and 1.0~30 mg/L, respectively. The relative standard deviation was 1.1%~2.0% and the recovery was 96.7%~103.2 % (n=6). The detection limits of BSA and HAS were 0.32 mg/L and 0.33 mg/L, respectively. The method was characterized by good recurrence, rapidity of oparation reaction, good stability and few infering substances. The results of the proteins in human serum samples were very close to those obtained by biuret spectrophotometric method.