López M Sánchez-Paniagua, Mecerreyes D, López-Cabarcos E, López-Ruiz B
Departamento de Físico-Química Farmacéutica, Facultad de Farmacia, Universidad Complutense de Madrid, 28040 Madrid, Spain.
Biosens Bioelectron. 2006 Jun 15;21(12):2320-8. doi: 10.1016/j.bios.2006.02.019. Epub 2006 Apr 17.
A glucose amperometric biosensor based on the immobilization of glucose oxidase (GOx) in microparticles prepared by polymerization of the ionic liquid 1-vinyl-3-ethyl-imidazolium bromide (ViEtIm+ Br-) using the concentrated emulsion polymerization method has been developed. The polymerization of the emulsion dispersed phase, in which the enzyme was dissolved together with the ionic liquid monomer, provides poly(ViEtIm+ Br-) microparticles with entrapped GOx. An anion-exchange reaction was carried out for synthesizing new microparticles of poly(ViEtIm+ (CF3SO2)2N-) and poly(ViEtIm+ BF4-). The enzyme immobilization method was optimized for biosensor applications and the following optimal values were determined: pH 4.0 for the synthesis medium, 1.23 M monomer concentration and 3.2% (w/w) cross-linking content. The performance of the biosensor as a function of some analytical parameters such as pH and temperature of the measuring medium, and enzymatic load of the microparticles was also investigated. The effect of the substances which are present in serum samples such as uric and ascorbic acid was eliminated by using a thin Nafion layer covering the electrode surface. The biosensor thus prepared can be employed in aqueous and in non-aqueous media with satisfactory results for glucose determination in human serum samples. The useful lifetime of this biosensor was 150 days.
基于使用浓乳液聚合法将葡萄糖氧化酶(GOx)固定在由离子液体1-乙烯基-3-乙基咪唑溴化物(ViEtIm⁺Br⁻)聚合制备的微粒中,开发了一种葡萄糖安培生物传感器。乳液分散相的聚合过程中,酶与离子液体单体一起溶解,从而得到包埋有GOx的聚(ViEtIm⁺Br⁻)微粒。进行了阴离子交换反应以合成聚(ViEtIm⁺(CF₃SO₂)₂N⁻)和聚(ViEtIm⁺BF₄⁻)的新微粒。针对生物传感器应用优化了酶固定方法,并确定了以下最佳值:合成介质的pH为4.0,单体浓度为1.23 M,交联含量为3.2%(w/w)。还研究了生物传感器的性能与一些分析参数的关系,如测量介质的pH和温度以及微粒的酶负载量。通过在电极表面覆盖一层薄的Nafion层,消除了血清样品中存在的尿酸和抗坏血酸等物质的影响。如此制备的生物传感器可用于水性和非水性介质中,对人血清样品中的葡萄糖进行测定,结果令人满意。该生物传感器的使用寿命为150天。