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为了检测神经毒剂类似物甲基膦酸二甲酯(DMMP),制备了一种新型的固定化植物酯酶-单磺酸基四苯基卟啉(TPPS1)纳米传感膜.首先对膜材制备的影响因素,如戊二醛浓度、活化时间和吸附时间等进行了优化研究.再通过原子力显微镜(AFM)和UV-Vis光谱对在优化条件下制备的膜材的表面形貌和对DMMP的传感性能分别进行了表征和分析.结果表明,当戊二醛浓度为0.43mol/L,活化时间为80min,吸附时间为170min时,固定化植物酯酶的活力较高.该膜材的纳米结构增加了其对DMMP的敏感性,降低了检测限.TPPS1与固定化植物酯酶作用后会在422nm处形成一特征峰.把固定化植物酯酶-TPPS1表面暴露于DMMP中,由于DMMP可以把卟啉从酶的活性位点上置换下来,所以引起422nm处吸光度的降低.并且,在DMMP浓度低于9×10-7mol/L的范围内,422nm处吸光度的变化与DMMP的浓度呈线性相关.该纳米传感膜可以检测浓度低至4.5×10-10mol/L的DMMP.
In order to detect the neurotoxic agent analog DMMP, a novel immobilized plant esterase-monosulfonate tetraphenylporphyrin (TPPS1) nanosensor was prepared.At first, Such as the concentration of glutaraldehyde, the activation time and the adsorption time, were optimized by atomic force microscopy (AFM) and UV-Vis spectroscopy.Furthermore, the surface morphology of the prepared membrane under optimized conditions and the effect of DMMP The results showed that when the concentration of glutaraldehyde was 0.43mol / L, the activation time was 80min and the adsorption time was 170min, the activity of immobilized esterase was high. The nanostructures increased their sensitivity to DMMP and decreased the detection limit.TPPS1 formed a characteristic peak at 422 nm after immobilized plant esterase.The surface of immobilized plant esterase-TPPS1 was exposed to DMMP, and DMMP The porphyrin can be displaced from the active site of the enzyme, resulting in a decrease in absorbance at 422 nm, and the change in absorbance at 422 nm and the concentration of DMMP in the range of DMMP concentration lower than 9 × 10 -7 mol / L were The nanosensor membranes can detect concentrations as low as 4 .5 x 10-10 mol / L DMMP.