罗丹明6G发光分子纳米微球标记凝集素-亲和吸附固体基质室温燐光法测定碱性磷酸酶

在外重原子微扰剂Pb（Ac）2存在下,基于罗丹明6 G（Rhodamine 6G,简写为R）的SiO2纳米微球（简写为R-SiO2）在醋酸纤维素膜（ACM）上于λex/λem=482.38/648.59 nm处能发射强而稳定的室温燐光信号.由该发光微球标记的麦胚凝集素（Triticum vulgare lectin简称WGA）,不仅仍能在ACM上与碱性磷酸酶（AP）发生定量的特异性亲和吸附反应,且亲和吸附反应产物能发射更强的室温燐光（RTP）信号,据此建立了R-SiO2纳米微球标记凝集素（WGA）亲和吸附（AA）固体基质室温燐光法（AA-SS-RTP）测定AP的一种新方法.本法的线性范围为1.00～360.00 ag AP spot^-1（样品体积0.4μL/斑,对应浓度2.50～900.00fg/ml））,工作曲线的回归方程为△Ip=16.24＋0.8856 m AP/ag spot^-1,r=0.9993.检出限为0.14ag spot^-1.分别对1.00和360.00 ag spot^-1 AP重复测定11次,RSD为3.9%和3.1.%.本方法灵敏、准确、精密度好,已成功用于人血清中AP的测定.

Affinity Adsorption Solid Substrate-room Temperature Phosphorimetry for the Determination of Alkaline Phosphatase

In the presence of Pb（Ac）2,the silicon dioxide nanoparticle containing rhodamine 6G（R-SiO2） can emit strong and stable solid substrate-room temperature phosphorescence（RTP） signal on the surface of acetyl cellulose membrane（ACM） at λex/λem = 482/649 nm.It was found in the research that specific affinity adsorption reaction between triticum vulgare lectin（WGA）（which was labeled with luminescent silicon dioxide nanoparticle） and alkaline phosphatase（AP） can be carried out on the surface of ACM.And the product of the reaction can emit stronger RTP signal.A new affinity absporption solial substrate room temperature phosphorimetry for the determination of AP was established,based on an affinity adsorption reaction between AP and WGA labeled with nanoparticles containing rhodanime 6G luminescent molecules.The linear range of this WGA-AP-WGA-R-SiO2 method is 1.00-360.00 ag AP spot^-1（sample volume： 0.40 μL spot^-1,corresponding concentration range： 2.50-900.00 fg m^l-1）.The regression equation of working curve is △Ip = 16.24 ＋ 0.8856 mAP（ag spot^-1）,r = 0.9993.Detection limit of this method calculated by 3Sb/k is 0.14 ag spot^-1.After eleven fold replicate measurements,RSD are 3.9℅ and 3.1℅ for the systems containing 1.00 and 360.00 ag AP spot^-1,respectively.Compared with R-SiO2-WGA-AP method（detection limit： 0.45 ag spot^-1,corresponding concentration range： 2.00-320.00 ag spot^-1）,the sensitivity of WGA-AP-WGA-R-SiO2 method was improved obviously and the linear range was wider.The sensitivity,accuracy and precision of this method are high.It has been applied to determine AP in human serum successfully.