Optical biochemical sensor for determining hydroperoxides in nonpolar organic liquids as archetype for sensors consisting of amphiphilic conetworks as immobilisation matrices
Optical biochemical sensor for determining hydroperoxides in nonpolar organic liquids as archetype for sensors consisting of amphiphilic conetworks as immobilisation matrices
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DOI：
10.1007/s00216-006-0680-2
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发表时间：
2006-11-01
影响因子：
4.3
通讯作者：
Heinze, Juergen
中科院分区：
文献类型：
--
作者：
Hanko, Michael;Bruns, Nico;Heinze, Juergen
本文报道了一种用于非极性有机液体中过氧化氢测定的光学生化传感器样机的成功设计。该传感器由两亲性聚合物网络（APCN）的基质组成，APCN是一类非常有前途的新型聚合物材料，易于制备生物化学传感器基质。APCN的特征在于亲水相和疏水相之间的纳米级相分离。对于中等比例的网络组合物，两相的域在网络的表面上和整个本体上都互连。APCNs具有独特的溶胀特性，亲水相在亲水介质中溶胀，疏水相在疏水介质中溶胀。在这两种类型的介质中，溶解的试剂可以从溶液扩散到聚合物网络的溶胀相中。这使得能够通过简单的浸渍用酶和指示剂加载APCN的亲水相。疏水分析物可以通过其疏水相扩散到聚合物网络中，并在两个相反相之间的巨大内部界面处与固定在亲水相中的指示剂反应。我们使用由58%（w/w）聚（丙烯酸2-羟乙酯）（PHEA）作为亲水链和42%（w/w）聚二甲基硅氧烷（PDMS）作为疏水连接体。辣根过氧化物酶（HRP）和作为指示剂的2，2 '-连氮基-双（3-乙基苯并噻唑啉-6-磺酸盐）二铵（ABTS）共固定在这种光学透明和透明的基质中。在该可行性研究中，研究的条件主要是与创新基质材料和由其生产的一次性生物传感器的表征相关的条件;生物传感器未优化。对叔丁基过氧化氢（tBuOOH）溶解在正庚烷中的灵敏度是可以接受的，约1和至少50 mmol L-1之间，即使在干燥状态。响应时间为1.7至5.0分钟。在至少1小时的时间内未观察到固定化试剂的沥滤。预溶胀的传感器与水增加的反应速率和总营业额的酶。在4摄氏度的干燥气氛中，传感器被发现可以稳定至少两周。
This paper reports the successful design of a prototype of an optical biochemical sensor for the determination of hydroperoxides in nonpolar organic liquids. The sensor consists of a matrix of an amphiphilic polymer conetwork (APCN), a novel class of very promising polymeric materials for easy preparation of biochemical sensor matrices. APCNs are characterised by nanoscopic phase separation between the hydrophilic and the hydrophobic phases. For medium ratios of conetwork composition, the domains of both phases are interconnected both on the surface of the conetworks and throughout the bulk. The APCNs have peculiar swelling properties-the hydrophilic phase swells in hydrophilic media and the hydrophobic phase swells in hydrophobic media. In both types of media dissolved reagents can diffuse from the solution into the swollen phase of the polymeric conetwork. This enables loading of the hydrophilic phase of the APCNs with enzymes and indicator reagents by simple impregnation. Hydrophobic analytes can diffuse into the polymeric conetwork via its hydrophobic phase and react with indicator reagents immobilised in the hydrophilic phase at the huge internal interface between the two opposite phases.To prepare the described hydroperoxide-sensitive biosensors, we used APCN films consisting of 58% (w/w) poly(2-hydroxyethyl acrylate) (PHEA) as hydrophilic chains and 42% (w/w) polydimethylsiloxane (PDMS) as hydrophobic linkers. Horseradish peroxidase (HRP) and diammonium 2,2 '-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) as indicator reagent were co-immobilised in this optically clear and transparent matrix. In this feasibility study the conditions investigated were principally those relevant to characterisation of the innovative matrix material and the disposable biosensor produced from it; the biosensor was not optimised. Sensitivity toward tert-butylhydroperoxide (tBuOOH) dissolved in n-heptane was acceptable, between approximately 1 and at least 50 mmol L-1, even in the dry state. The response time was 1.7 to 5.0 min. No leaching of immobilised reagents was observed during a period of at least one hour. Pre-swelling the sensors with water increased the reaction rate and the total turnover number of the enzyme. In a dry atmosphere at 4 degrees C the sensors were found to be stable for at least two weeks.