5'腺苷单磷酸钠探针同步荧光测定生物样品中蛋白质含量

5'腺苷单磷酸钠(AMPNa)与人血清白蛋白(HSA)相互结合可以形成没有荧光的复合物,进而导致HSA的内源荧光发生改变,并且体系的同步荧光强度和溶液中的HSA浓度呈现出良好的线性关系.根据这些现象,建立了同步荧光光谱法测定生物样品中蛋白质含量的新方法,而且这种方法是把AMPNa作为分子探针的.实验中考察了一些因素对同步荧光光谱特征及强度的影响,这些因素包括Δλ值、反应介质及温度等.AMPNa HSA的浓度在1.43~244.8μg·mL-1范围内的同步荧光强度与蛋白质的浓度呈现出较好的线性关系.在较为理想的实验条件下,对11份空白溶液进行平行测定,检测限可达到0.319μg·mL-1,并对血清、尿样和唾液中的蛋白质含量进行测定并进行加标回收实验,回收率在95.06%~104.89%.

Determination of Proteins Content in Biology Samples with Adenosine 5'-monophosphate sodium salt as Probe by Synchronous Fluorescence Technique

The interaction of 5'-monophosphate sodium salt(AMPNa)and human serum albumin(HSA)can form a kind of complex substances with no fluorescence.This can lead to change of the fluorescence of HSA.There is a linear relation between the intensity of synchronous fluorescence and the concentration of HSA.Then we establish a new method for the determination of proteins using the AMP-Na as probe by synchronous fluorescence technique.In this experiment,we investigate some factors includeΔλ,reaction medium and temperature on the synchronous fluorescence intensity.Under the optimum experimental conditions,the synchronous fluorescence intensities of AMPNa was in good proportion to the concentration of HSA in the range of 1.43~244.8μg·mL-1.The detection limit is 0.319μg·mL-1 by determination of 11blank solution.Determination of protein in serum,urine and saliva samples under the optimum conditions,the recovery is in the range 95.06%~104.89%.