一种新的室温燐光法及其分析应用

A New Room Temperature Phosphorimetry for Determination of Trace Phosphatase and its Analytical Application

提出一种新的4代树枝状分子(4G-D)-曲拉通-100X(Triton-100X)磷光标记试剂.基于Triton-100X-4G-D标记的麦胚凝集素(WGA)的产物(Triton-100X-4G-D-WGA),能与碱性磷酸酶(ALP)发生两种(直接法和夹心法)特异性亲和吸附反应(AA),其AA的产物仍然保持4G-D室温磷光的优良性质,且AA产物的-Ip值与ALP的含量成线性关系,据此建立了Triton-100X-4G-D标记麦胚凝集素-亲和吸附固体基质室温燐光法(AA-SS-RTP)测定痕量ALP的新方法.该方法的检出限为0.20agspot-1(直接法,对应浓度:5.0×10-18mol L-1)、0.14agspot-1(夹心法,对应浓度:3.5×10-18mol L-1),灵敏度高,重现性和精密度好.直接法和夹心法用于血清中ALP含量的测定,与漳州市中医院ELISA法临床检测的结果相吻合.

A Triton X-100-4.0G-D （4.0G-D refers to a 4.0-generation dendrimer） was brought forward as a new phosphorescence labeling reagent. Two types of specific affinity adsorption （AA） reactions （direct method and sandwich method） were carried out between the labeling product of Triton X-100-4.0G-D-Wheat germ agglutinin （WGA） and alkaline phosphatase （ALP）, the product of AA reaction preserved the good characteristics of room temperature phosphorescence （RTP） of 4.0G-D and AIP of the product was proportional to the content of ALP. According to the fact stated above, a new method for the determination of trace ALP by affinity adsorption solid substrate-room temperature phosphorimetry （AA-SS-RTP） was established on the basis of WGA labeled with the Triton X-100-4.0G-D. The detection limits were 0.20 agospot-1 （corresponding concentration： namely 5.0×10^-18 tool-L^-1） for a direct method and 0.14 ag·spot-1 （corresponding concentration： namely 3.5×10^-18 mol·L^-1） for a sandwich method, respectively. For their high sensitivity, good repeatability and high accuracy, the direct method and sandwich method have been successfully applied to determine the content of ALP in human serum, and the results were coincided with the clinical detection results of the enzyme-linked immunosorbent assay method by the Zhangzhou Hospital of Traditional Chinese Medicine.