卟啉全内反射同步荧光法测定蛋白质

Determination of Protein with TPPS by Total Internal Reflection Synchronous Fluorescence Spectroscopy

应用全内反射与同步荧光相结合的技术,建立了利用蛋白质吸附在液-固界面上的同步荧光信号来 测定本体蛋白质溶液浓度的新方法。其原理为mesi-四(4-磺酸基苯基)卟啉(TPPS)标记牛血清蛋白(BSA)在 石英界面的吸附后信号强度随本体溶液浓度的增加呈线性关系。方法的检出限是94 ng·mL-1。用于实际人 血清蛋白样品的测定,结果令人满意。

A new method of quantitative detemaination for serum albumin in aqueous solution tins been developed by measuring total internal reflection synchronous fluorescence at the solid/liquid interface. The combination of bovine serum album in （BSA） and mesotetrakis （4-sulphonatophenyl） porphyrin （TPPS） adsorbed onto the glass surface produced a synchronous fluorescence signal at 421 nm. At pH 4.25, the signal intensity of BSA adsorbed on the interface was proportional to the BSA concentration in bulk solution. The linear range of 1.0-8.0 μg· mL^-1 and the detection limit of 0.94 μg· mL^-1 were obtained. The human serum samples were determined with satisfactory results.