含铁（Ⅲ）纳米材料的模拟酶活性研究及其分析应用

发布时间：2018-05-17 01:41

本文选题：纳米模拟酶 + 金属有机框架化合物　；参考：《西南大学》2017年硕士论文

【摘要】：生物酶具有高催化活性和特异性,但由于提取困难、纯化复杂、稳定性差、需要温和的反应条件等缺点,导致其应用受到限制。纳米材料因具备独特的物理化学性质,与生物酶的尺寸、形状、催化活性等具有相似性而被广泛研究。近年来对碳纳米材料、金属纳米粒子和金属氧化物纳米粒子等纳米模拟酶的研究越来越多。金属有机框架化合物(MOFs)和层状双金属氢氧化物(LDHs)都具有金属离子可调,配合物或阴离子可改变的性质。此外,未配位饱和的金属离子是催化反应的活性中心,而Fe(Ⅲ)的催化活性是金属离子中较为突出的。因此,本文对近年来纳米材料模拟酶和提高纳米材料催化活性的方法进行了总结;对含铁的MOFs和LDHs的模拟酶活性进行了研究,并以化学发光法和比色法两种手段建立了快速灵敏地检测H_2O_2和葡萄糖浓度的分析方法。首先,本文利用含铁的金属有机框架化合物(MOFs),即MIL-53(Fe)大幅度增强了luminol-H_2O_2体系在碱性反应条件下的化学发光,其强度是luminol-H_2O_2体系的20倍。XRD证明MIL-53(Fe)反应前后的XRD形态没有发生变化。通过对该化学发光增强体系的发光强度,荧光波谱和活性氧自由基的捕获实验等的分析,结果表明MIL-53(Fe)在MIL-53(Fe)-luminol-H_2O_2体系中扮演催化剂的角色且具有较好的稳定性,化学发光增强的原因可能是体系中生成了大量的O_2?-和?OH等活性自由基。结合葡萄糖氧化酶催化氧化葡萄糖产生H_2O_2的反应,我们建立了一种灵敏的、高选择性的检测葡萄糖的方法。化学发光强度的对数值和葡萄糖浓度的对数值之间存在良好的线性关系,线性范围为0.1-10μM,检出限为0.05μM。该方法在检测血清中葡萄糖浓度时得到了令人满意的结果。其次,本文还发现花球状CoFe LDH对luminol-H_2O_2体系和luminol体系的化学发光都有很强的促进作用,化学发光强度分别增强了180倍和14倍。机理研究表明,luminol-H_2O_2-CoFe LDH体系生成了大量的?OH和O_2?-;luminol-Co Fe LDH体系中只生成了大量的O_2?-。活性氧中间体自由基与luminol反应,促进了化学发光强度的增强。基于CoFe-LDH对luminol-H_2O_2体系化学发光强度的增强效应,本文建立了一种快速简单灵敏的测定H_2O_2和葡萄糖浓度的方法。对H_2O_2的线性范围为0.01-3μM,检出限为5 nM;对葡萄糖的线性范围为0.07-3μM,检出限为50 nM。我们还利用抗坏血酸清除O_2?-抑制luminol-Co Fe LDH体系化学发光的原理,建立了快速测定抗坏血酸的化学发光分析方法。最后,我们考虑到纳米材料的形貌会对其催化活性产生影响。因此,本文还制备了纳米片、纳米卷和纳米花三种不同形貌的NiFe LDH,并对它们的过氧化物模拟酶活性进行了研究。经SEM、XRD、红外光谱和热重分析等表征手段证明三种不同形貌的NiFe LDH被成功制备。相比于NiFe LDH纳米片和纳米卷,柠檬酸钠插入的花球状t-NiFe-LDH-SC(0.4)具有更高的过氧化物模拟酶活性;同时花球状的t-NiFe-LDH-SC(0.4)具有对H_2O_2最高的亲和力和对3.3′,5.5′-四甲基联苯胺(TMB)有最大的反应速率。与已有的纳米材料模拟酶相比,t-NiFe-LDH-SC(0.4)也表现出对H_2O_2非常高的亲和力。柠檬酸钠的插入为催化反应提供了更多的结合位点和催化位点。t-NiFe-LDH-SC(0.4)催化TMB-H_2O_2的显色反应的动力学符合米氏动力学,反应机理遵循乒乓机理。基于花球状t-Ni Fe-LDH-SC(0.4)的过氧化物模拟酶活性,本文建立了一种简单,快速检测过氧化氢和葡萄糖浓度的方法。
[Abstract]:Biological enzymes have high catalytic activity and specificity, but because of the difficulties in extraction, complex purification, poor stability and the need for mild reaction conditions, their applications are restricted. Nano materials have been widely studied because of their unique physical and chemical properties, similar to the size, shape, and catalytic activity of biological enzymes. Nanomaterials, metal nanoparticles and metal oxide nanoparticles have been studied more and more. Both metal organic frame compounds (MOFs) and layered double metal hydroxide (LDHs) have the properties of adjustable metal ions, complexes or anions. In addition, the activity of unmatched saturated metal ions is the activity of catalytic reaction. The catalytic activity of Fe (III) is more prominent in metal ions. Therefore, in this paper, the methods of simulating enzymes and improving the catalytic activity of nanomaterials in recent years were summarized. The activity of simulated enzyme in MOFs and LDHs containing iron was studied. The rapid and sensitive detection was established by two means of chemical hair light and colorimetric method. The analysis method for measuring H_2O_2 and glucose concentration. First, in this paper, the chemiluminescence of the luminol-H_2O_2 system under the alkaline reaction condition was greatly enhanced by the metal organic frame compound (MOFs) containing iron, that is, MIL-53 (Fe). The strength of the luminol-H_2O_2 system was 20 times of the luminol-H_2O_2 system, and the XRD morphology before and after the MIL-53 (Fe) reaction did not change. The results show that MIL-53 (Fe) plays the role of catalyst in MIL-53 (Fe) -luminol-H_2O_2 system and has good stability by the analysis of the luminescence intensity of the chemiluminescence enhancement system, the fluorescence spectrum and the capture experiment of reactive oxygen free radicals. The reason for the enhancement of chemiluminescence may be the formation of a large number of O_2? - and OH in the system. In conjunction with glucose oxidase catalyzed oxidation of glucose to produce H_2O_2, we have established a sensitive, highly selective method for the detection of glucose. There is a good linear relationship between the value of the chemiluminescence intensity and the value of glucose concentration, the linear range is 0.1-10 M, the detection limit is 0.05 u M.. The method obtained satisfactory results in detecting the glucose concentration in serum. Secondly, this paper also found that the flower spherical CoFe LDH has a strong effect on the chemiluminescence of the luminol-H_2O_2 system and the luminol system, and the chemiluminescence intensity is increased by 180 times and 14 times respectively. The mechanism research shows that the luminol-H_2O_2-CoFe LDH system has been formed. A large number of OH and O_2? - luminol-Co Fe LDH systems have produced only a large number of O_2? - the free radicals of reactive oxygen intermediates and luminol reacted to promote the enhancement of chemiluminescence intensity. Based on the enhancement effect of CoFe-LDH on the chemiluminescence intensity of luminol-H_2O_2 system, a rapid and simple and sensitive determination of H_2O_2 and glucose concentration was established. The linear range of H_2O_2 is 0.01-3 mu M, the detection limit is 5 nM, the linear range of glucose is 0.07-3 mu M, the detection limit is 50 nM., and we also use ascorbic acid to clear O_2? - the principle of inhibiting luminol-Co Fe LDH system chemiluminescence, and establish a chemiluminescence analysis method for rapid determination of ascorbic acid. Finally, we consider nano The morphology of rice materials will affect its catalytic activity. Therefore, the NiFe LDH of three different morphologies of nanoscale, nanoscale and nanoscale were prepared and their peroxidase activity was studied. The three different morphologies of NiFe LDH were successfully prepared by means of SEM, XRD, infrared spectroscopy and thermogravimetric analysis. Compared to NiFe LDH nanoscale and nanoscale, the flower ball like t-NiFe-LDH-SC (0.4) inserted by sodium citrate has higher activity of peroxidase analog enzyme; at the same time, the spherical t-NiFe-LDH-SC (0.4) has the highest affinity for H_2O_2 and the maximum reaction rate for 3.3 ', 5.5' - four methylbiphenyl amine (TMB). Compared with the enzyme, t-NiFe-LDH-SC (0.4) also showed a very high affinity for H_2O_2. The insertion of sodium citrate provided a more binding site and a catalytic site.T-NiFe-LDH-SC (0.4) to catalyze the chromogenic reaction of TMB-H_2O_2, which conforms to the Micheli dynamics, and the reaction mechanism follows the table table mechanism. Based on the flower like t-Ni Fe-LDH-SC (0.) 4) the activity of peroxide mimetic enzyme. A simple and rapid method for the determination of hydrogen peroxide and glucose concentration was established.
【学位授予单位】：西南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.3;O629.8

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