葡萄糖的酶法测定方法详细操作事项
来源：http://www.jnjdbc.com 日期：2019-07-11 发布人：admin
葡萄糖氧化酶法葡萄糖氧化酶法原理如下： 葡萄糖被葡萄糖氧化酶氧化生成葡萄糖酸和H2O2，H2O2又经辣根过氧化物酶作用，合成出氧，将无色的4-氨基安替比林和苯酚偶联氧化，并缩合成红色醌亚胺，其颜色深浅与葡萄糖浓度呈正比，在530 nm下的吸收峰度值会随葡萄糖浓度的加而增加。应用这一原理能够测定葡萄糖含量。
The principle of glucose oxidase method is as follows: Glucose is oxidized by glucose oxidase to produce gluconic acid and hydrogen peroxide, and then hydrogen peroxide reacts with horseradish peroxidase to synthesize oxygen. Colorless 4-aminoantipyrine and phenol are oxidized by coupling, and red quinone imide is synthesized by condensation. In direct proportion, the absorption peak at 530 nm increases with the increase of glucose concentration. This principle can be used to determine the content of glucose.
葡萄糖氧电极法：葡萄糖氧电极法的原理是采用氧耗费速率检测葡萄糖，将氧电极置于含有适量葡萄糖氧化酶的溶液中，然后参加待测样品，样品中的葡萄糖被氧化而耗费氧。由于氧耗费量与血糖浓度呈正比，而电极的极限扩散电流又与溶液中的氧含量呈正比，因而，氧电极值即可反映样品中血糖浓度。采用电化学聚合技术，用掺杂苯磺酸钠的聚吡咯( PPy) 导电薄膜修饰铅笔芯电极，在修饰电极外表吸附葡萄糖氧化酶制备了葡萄糖生物传感器。该生物传感器抗干扰才能强、稳定性好，响应电流和葡萄糖浓度在0～0.7 mmol /L 范围内有良好的线性相关度，灵活度为26.10 uA/mmol /L，均匀响应时间约为6.5 s，检测下限为47.2 mol /L。
Glucose Oxygen Electrode Method: The principle of Glucose Oxygen Electrode Method is to use oxygen consumption rate to detect glucose. First, the oxygen electrode is placed in a solution containing appropriate amount of glucose oxidase, and then participates in the sample to be tested. The glucose in the sample is oxidized and consumes oxygen. Oxygen consumption is proportional to blood sugar concentration, and the limiting diffusion current of the electrode is proportional to the oxygen content in the solution. Therefore, the oxygen electrode value can reflect the blood sugar concentration in the sample. The pencil-core electrode was modified by polypyrrole (PPy) film doped with sodium benzenesulfonate by electrochemical polymerization. Glucose biosensor was prepared by adsorbing glucose oxidase on the surface of the modified electrode. The biosensor has strong anti-jamming ability and good stability. The response current and glucose concentration have good linear correlation in the range of 0-0.7 mmol/L. The flexibility is 26.10 uA/mmol/L, the uniform response time is about 6.5 s, and the detection limit is 47.2 mol/L.
纳米资料模仿酶比色法：葡萄糖氧化酶传感器检测葡萄糖浓度，具有专注性高、反响速度快等特性。但酶自身固有的不稳定性，易受温度、湿度以及pH 值等环境条件的影响而失去活性，而且葡萄糖氧化酶价钱昂贵、制备复杂，这在一定水平上限制了酶传感器的应用。应用具备催化活性的纳米资料，可以研制无酶葡萄糖传感器。基于CeO2NPs 催化TMB 显色反响与H2O2浓度的线性关系，构建了测定血样中葡萄糖的模仿酶比色剖析检测办法。他们合成了一种稳定和水溶性的聚丙烯酸修饰的CeO2NPs，发现CeO2 NPs 可以催化H2O2氧化，表现出过氧化物模仿酶的催化活性。在优化条件下，对葡萄糖的线性响应范围为0.5～10 mmol /L，检出限为0.1 mmol /L。对1.0 mmol /L 葡萄糖停止11 次平行测定，其相对规范偏向为2.4 %。即便果糖、乳糖和麦芽糖的浓度高达5 mmol /L，也不会干扰葡萄糖的测定，具有很高的选择性。
Nanodata mimic enzyme colorimetry: Glucose oxidase sensor detects the concentration of glucose, which has the characteristics of high concentration, fast response and so on. However, the inherent instability of the enzyme itself, which is vulnerable to temperature, humidity and pH and other environmental conditions, makes it inactive. Moreover, the cost of glucose oxidase is expensive and its preparation is complex, which limits the application of the enzyme sensor to a certain extent. Enzyme-free glucose sensor can be developed by using nano-materials with catalytic activity. Based on the linear relationship between the color reaction of TMB catalyzed by CeO2 NPs and the concentration of hydrogen peroxide, a colorimetric analysis method for the determination of glucose in blood samples by mimetic enzyme was constructed. They first synthesized a stable and water-soluble polyacrylic acid-modified CeO_2 NPs, and found that CeO_2 NPs could catalyze the oxidation of H_2O_2, showing the catalytic activity of peroxidase mimicking enzyme. Under the optimum conditions, the linear response range for glucose was 0.5-10 mmol/L, and the detection limit was 0.1 mmol/L. For 1.0 mmol/L glucose, 11 parallel measurements were stopped, and the relative standard deviation was 2.4%. Even if the concentration of fructose, lactose and maltose is as high as 5 mmol/L, it will not interfere with the determination of glucose and has high selectivity.
无酶型葡萄糖电化学法：无酶葡萄糖电化学法也是不运用葡萄糖氧化酶的葡萄糖测定办法，具有较高的稳定性、良好的重现性和本钱低廉等特性，近年来得到了快速开展。目前，曾经有单一金属( Pt，Pd，Au 等) 、双金属( Pt-Au，Pt-Pb，Pt-Ru，Pt-Bi，Pt-Ti 等) 及碳纳米管、碳纤维、介孔碳等纳米资料被用于无酶型葡萄糖电化学传感器的研制。经过电化学堆积的办法将Pt-Pd 双金属纳米粒子堆积在裂解碳纳米管修饰的电极外表上，制备了Pt-Pd /UCNTs 无酶葡萄糖传感器。该传感器对0～20 mmol /L浓度范围内的葡萄糖有良好的响应，不受Cl u等物质的干扰，表现了良好的选择性。
Enzymatic-free glucose electrochemical method: Enzymatic-free glucose electrochemical method is also a glucose determination method without using glucose oxidase. It has high stability, good reproducibility and low cost, and has been developed rapidly in recent years. At present, single metal (Pt, Pd, Au, etc.), bimetal (Pt-Au, Pt-Pb, Pt-Ru, Pt-Bi, Pt-Ti, etc.) and carbon nanotubes, carbon fibers and mesoporous carbon have been used in the development of enzyme-free glucose electrochemical sensors. Pt-Pd/UCNTs enzyme-free glucose sensor was prepared by electrochemical deposition of Pt-Pd bimetallic nanoparticles on the surface of the electrode modified by pyrolysis carbon nanotubes. The sensor has a good response to glucose in the concentration range of 0-20 mmol/L, and is not disturbed by Cl u and other substances, showing good selectivity.
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