微乳液增敏高灵敏荧光光谱探针测定蛋白质的研究

Microemulsion Sensitized Fluorophotometric Determination of Protein with Probe of Phenylfluorone-Molybdenum(Ⅵ) Complex

研究了一种新颖的测定蛋白质的荧光光谱探针,在TritonX100微乳液存在下,pH2.0的HClKCl介质中,考察了钼(Ⅵ)苯基荧光酮(PF)蛋白质体系的荧光光谱特性,该体系的最大激发波长和最大发射波长分别为465和525nm。在适宜的实验条件下,蛋白质的浓度在以下浓度范围内与ΔF呈良好的线性关系:0～6.00μg·L-1牛血清蛋白,0～4.00μg·L-1人血清蛋白;0～5.00μg·L-1卵清蛋白;0～4.00μg·L-1溶菌酶。将TritonX100微乳液引入到蛋白质的测定中,改善了体系的微环境,显著地提高了测定的灵敏度,检测限分别为5.4,5.2,1.5和8.2ng·L-1。实验结果表明:该法具有很高的灵敏度、选择性和稳定性,可直接用于尿样的定量测定。

The fluorescence spectral behavior of interaction of phenylfluorone （PF）-Mo （ Ⅵ ） and protein was investigated in Triton X-100 mieroemulsion medium at pH 2.0. A novel method for the determination of protein using phenylfluorone （PF）-Mo（ Ⅵ） as a fluorescence spectrum probe was developed. Excitation and emission wavelengths are 465 and 525 nm, respectively. The effective factors and the optimum conditions have been studied, and the reducing value of fluorescence intensity is in proportion to the concentration of proteins in the range 0-6.00μg·L^-1 for bovine .serum albumin, 0-4.00 μg·L^-1 for human serum albumin, 0-5.00 μg · L^-1 for ovalbumin, and 0-4.00 μg·L^-1 for lysozyme. The Triton X-100 microemulsion was efficiently used to enhance the sensibility and stability of the system, and the limits of detection are 5.4, 5.2, 1.5 and 8.2 ng· L^- 1, respectively. Most of foreign suhstances do not interfere with the determination, and this method has good selectivity and high sensitivity. It has been applied to the determination of proteins in the urine samples with satisfactory results.