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β-环糊精修饰载体制备含 PPO 的酚类化合物生物传感器及铱纳米粒子的发展。

Development of biosensor for phenolic compounds containing PPO in β-cyclodextrin modified support and iridium nanoparticles.

机构信息

Departamento de Química, Laboratório de Biossensores, Universidade Federal de Santa Catarina, 88040-970, Florianópolis, SC, Brazil.

出版信息

Enzyme Microb Technol. 2013 Apr 10;52(4-5):296-301. doi: 10.1016/j.enzmictec.2012.12.001. Epub 2013 Jan 23.

Abstract

A biosensor based on the iridium nanoparticles dispersed in ionic liquid (IL) 1-butyl-3-methylimidazolium hexafluorophosphate (Ir-BMI·PF6) and a celery (Apium graveolens) extract as a source of polyphenol oxidase (PPO) was constructed. A modified support based on β-cyclodextrin (β-CDEP) was used for enzyme immobilization. The behavior of phenolic compounds was investigated by square-wave voltammetry and rutin was selected by presenting the greatest signal. The best performance was obtained with a composition of 70:10:10:10% (w/w/w/w) of the graphite powder:β-CDEP:Nujol:Ir-BMI·PF6 composition, a PPO concentration of 500unitsmL(-1), in 0.1M phosphate buffer solution (pH 6.0) with frequency, pulse amplitude and scan increment at 100Hz, 60mV, and 3.0mV, respectively. Under optimized conditions, the cathodic currents increased linearly for the rutin concentration range of 1.3×10(-7)-2.0×10(-6)M with a detection limit of 7.9×10(-8)M. This sensor demonstrated acceptable repeatability and reproducibility and the results for the rutin recovery ranged from 92.8 to 103.4%. A relative error of 0.7% was obtained in the rutin determination in simulated samples.

摘要

基于离子液体(IL)1-丁基-3-甲基咪唑六氟磷酸盐(Ir-BMI·PF6)中分散的铱纳米粒子和芹菜(Apium graveolens)提取物作为多酚氧化酶(PPO)来源的生物传感器被构建。一种基于β-环糊精(β-CDEP)的改性载体被用于酶的固定化。通过方波伏安法研究了酚类化合物的行为,并选择芦丁作为信号最大的化合物。在最佳条件下,当石墨粉:β-CDEP:Nujol:Ir-BMI·PF6 的比例为 70:10:10:10%(w/w/w/w),PPO 浓度为 500unitsmL(-1),在 0.1M 磷酸盐缓冲溶液(pH 6.0)中,频率、脉冲幅度和扫描增量分别为 100Hz、60mV 和 3.0mV 时,传感器的性能最佳。在优化条件下,对于芦丁浓度范围为 1.3×10(-7)-2.0×10(-6)M 的电流响应呈线性增加,检测限为 7.9×10(-8)M。该传感器具有良好的重复性和重现性,芦丁回收率在 92.8%至 103.4%之间。在模拟样品中测定芦丁的相对误差为 0.7%。

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