DhHP-6替代过氧化物酶在葡萄糖检测中的应用

过氧化物酶（POD）是催化H2O2氧化反应的生物酶，很多涉及氧化反应的临床诊断试剂盒都采用这个原料。但天然的POD易失活、价格高、实验条件和储存环境都很苛刻。拟开发一种模拟酶替代过氧化物酶。次血红素六肽（DhHP-6）是一种生物小分子，由于它稳定、实验条件温和、成本较低等优点，成为近年关注热点。将次血红素六肽（DhHP-6）替代过氧化物酶催化H2O2氧化，通过偶联还原型色原N-乙基-N-（2-羟基-3-磺酸基丙基）-3，5-二甲氧基苯胺（DAOS）和4-氨基安替比林4-（AAP）进行显色反应，将该体系应用于葡萄糖氧化酶（GOD）催化体系，实现血液中葡萄糖含量的测定。实验结果表明，在最佳条件下，H2O2的线性范围为0.69~22.0mmol／L；葡萄糖测定的线性范围为0.35~22.0mmol/L。将本法用于血样中葡萄糖测定，重复性：CV=2.6%（ n=20）；稳定性：3小时内，浓度CV≤5.0%；与商品血清葡萄糖试剂进行比较，R≥0.98，平均回收率达到96.4％。结果为过氧化物酶的模拟酶研究与开发提供实验技术基础。

The Application of DhHP-6 Substitutes for Peroxidase in the Detection of Glucose

Peroxidase (POD) is a biological enzyme that catalyzes the oxidation of H2O2,that material of reagent kit re-late to oxidation. Conditions for natural POD were harsh that easily inactivity,high price,difficult experimental conditions and storage line. This study develop mimic enzyme instead of peroxidase. DhHP-6 is small molecule that became the hotspot for stability,safe experimental conditions and low cost. In this experiment,(DhHP-6) replace the peroxidase for catalyzed H2O2oxygen. The chromogenic reaction is through coupling N-Ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-di-methoxyaniline sodium salt (DAOS) and 4-Aminoantipyrine(4-APP). This system was applied to the glucose oxidase (GOD) catalytic system for determination of the blood glucose. Experimental results show that,under the optimal condi-tions,H2O2 linear region between 0.69~22.0mmol/L,Linear range of glucose determination between 0.35~22.0mmol/L. The method applied to determination of serum glucose,repeatability CV=2.6%( n=20),stability:concentration CV≤5.0%in 3 hours. Compared with commercial glucose kits,R≥0.98,the average recovery rate is 96.4%. It is helpful for mimic enzyme replaced peroxidase to develop and basical research.