基于蛋壳膜固定酶和纳米银膜光度法对葡萄糖的检测

利用蛋壳膜固定酶和静电作用组装纳米银膜,建立了一种新型的光度法检测微量葡萄糖.采用共价交联法在蛋壳膜上固定葡萄糖氧化酶.酶促反应产生的H_2O_2氧化纳米银导致纳米银膜吸光度减小,日光强度的变化值与葡萄糖浓度有关,据此实现了对葡萄糖的检测.该方法检测快速,线性范围为6.0×10~(-5)～66.0×10~(-5)mol/L(r=0.999 7,n=11),检出限(S/N=3)为1.8×10~(-5) mol/L.方法成功用于临床血清样品中葡萄糖含量的检测.

A Novel Spectrophotometric Method for Glucose Detection Based on Ag Nanoparticles Film and Glucose Oxidase-immobilized Eggmembrane

A novel spectrophotometric method has been developed for determination of glucose based on enzyme-immobilized eggshell membrane and Ag nanoparticles film.Ag nanoparticles film was prepared by a polyelectrolyte electrostatic assembly approach using poly(diallydimethethyl ammonium chloride)(PDDA)/polystyrene sodium sulfonate(PSS)/poly(vinylbenzyltrimethyl ammonium chloride)(PVTC) multilayers.Glucose oxidase was covalently immobilized on eggshell membrane using glutaraldehyde as cross-linking agents and was characterized by SEM.The detection scheme based on the depletion of Ag nanoparticles by H_2O_2 from exposure to glucose solution,and the decrease of the absorbance intensity related to the glucose concentrations.KCl was used to improve sensitivity and selectivity of the detection system.The effects of glutaraldehyde concentrations and pH were studied in details.Common matrix interferents such as ascorbic acid and uric acid did not give significant interference.Thus Ag nanopartilces film exhibited reliable reproducibility,excellent stability and fast response(2 min).The linear detection ranged from 6.0×10~(-5)to 66.0×10~(-5) mol/L (R=0.999 7,n=11) and the detection limit was 1.8×10~(-5) mol/L (S/N=3).Thus the method has been successfully applied for determination of glucose contents in serum samples.