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基于血红素的过氧化物酶样分子印迹聚合物的合成及其在血清中5-羟色胺选择性测定中的应用

Synthesis and application of a peroxidase-like molecularly imprinted polymer based on hemin for selective determination of serotonin in blood serum.

作者信息

Santos Wilney de Jesus Rodrigues, Lima Phabyanno Rodrigues, Tarley César Ricardo Teixeira, Höehr Nelci Fenalti, Kubota Lauro Tatsuo

机构信息

Departamento de Química Analítica, Instituto de Química, Universidade Estadual de Campinas, Campinas, SP 13084-971, Brazil.

出版信息

Anal Chim Acta. 2009 Jan 12;631(2):170-6. doi: 10.1016/j.aca.2008.10.034. Epub 2008 Nov 1.

Abstract

This work reports the preparation of a molecularly imprinted polymer (MIP) for selective catalytic detection of serotonin (5-hydroxytryptamine, 5-HT). The process is based on the synthesis of polymers with hemin introduced as the catalytic center to mimic the active site of peroxidase. The copolymer MIP, containing artificial recognition sites for 5-HT, has been prepared by bulk polymerization using methacrylic acid (MAA) and hemin as the functional monomers, and ethylene glycol dimethacrylate (EGDMA) as the cross-linker. For the determination of 5-HT, a flow injection analysis system coupled to an amperometric detector was optimized using multivariate analysis. The effects of different parameters, such as pH, buffer flow rate, buffer nature, peroxide concentration and sample volume were evaluated. After optimizing the experimental conditions, a linear response range from 1.0 up to 1000.0 micromolL(-1) was obtained with a sensitivity of 0.4nA/ micromolL(-1). The detection limit was found to be 0.30 micromolL(-1), while the precision values (n=6) evaluated by relative standard deviation (R.S.D.) were, respectively, 1.3 and 1.7% for solutions of 50 and 750 micromolL(-1) of 5-HT. No interference was observed by structurally similar compounds (including epinephrine, dopamine and norepinephrine), thus validating the good performance of the imprinted polymer. The method was applied for the determination of 5-HT in spiked blood serum samples.

摘要

本研究报道了一种用于选择性催化检测血清素(5-羟色胺,5-HT)的分子印迹聚合物(MIP)的制备方法。该过程基于合成以血红素作为催化中心的聚合物,以模拟过氧化物酶的活性位点。通过本体聚合,以甲基丙烯酸(MAA)和血红素作为功能单体,乙二醇二甲基丙烯酸酯(EGDMA)作为交联剂,制备了含有5-HT人工识别位点的共聚物MIP。为了测定5-HT,使用多变量分析对与安培检测器联用的流动注射分析系统进行了优化。评估了不同参数的影响,如pH值、缓冲液流速、缓冲液性质、过氧化物浓度和样品体积。优化实验条件后,获得了1.0至1000.0 μmolL⁻¹的线性响应范围,灵敏度为0.4 nA/μmolL⁻¹。检测限为0.30 μmolL⁻¹,而对于50和750 μmolL⁻¹的5-HT溶液,通过相对标准偏差(R.S.D.)评估的精密度值(n = 6)分别为1.3%和1.7%。未观察到结构相似化合物(包括肾上腺素、多巴胺和去甲肾上腺素)的干扰,从而验证了印迹聚合物的良好性能。该方法应用于加标血清样品中5-HT的测定。

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