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    基于聚多巴胺球负载Cu纳米粒子的催化作用分光光度法测定人血中葡萄糖含量

    Spectrophotometric Determination of Glucose in Blood Based on the Catalytic Reaction of Copper Nanoparticles on Polydopamine Spheres

    • 摘要: 用水热法在聚多巴胺球(PDA)表面负载了Cu纳米粒子(Cu@PDA),制备了具有过氧化物酶活性的Cu@PDA复合物。此复合物能催化过氧化氢快速氧化无色还原态的四甲基联苯胺(TMB),使其转变为蓝色的氧化态TMB,其吸收峰波长为652 nm,其吸光度与过氧化氢的浓度有关。利用此催化反应结合葡萄糖氧化酶(Gox)催化经氧气氧化葡萄糖而产生过氧化氢的反应,提出了分光光度法测定血样中葡萄糖的方法。在总体积为10 mL的0.1 mol·L-1磷酸盐缓冲溶液(pH 4.0)中含1.0 mg·L-1 Cu@PDA,4.0 g·L-1 Gox,0.5 mmol·L-1 TMB的条件下加入血样,于40 ℃反应10 min时,葡萄糖浓度在1.0~30.0 mmol·L-1的范围内与相应的吸光度之间呈线性关系,检出限(3S/N)为0.3 mmol·L-1。方法应用于人血样品的分析,所测得葡萄糖含量与医院实验室所给的值相符。

       

      Abstract: Copper nano particles were loaded on the surface of polydopamine spheres (PDA) by the hydrothermal method to give a composite of Cu@PDA which behave with activity of peroxidase and can catalyze the rapid oxidation of 3,3',5,5'-tetramethyl benzidine (TMB) by transforming from its colorless reduction form to its blue colored oxidation form, having its absorption maximum at 652 nm, and giving magnitude of absorbances measured related to the concentration of H2O2 in definite range. Applying this catalytic reaction in combination with the catalysis of oxidation reaction between oxygen and glucose to give H2O2 by glucoxydase (Gox), a spectrophotometric method for determination of glucose in blood samples was proposed. A definite amount of blood sample solution was added to 0.1 mol·L-1 PBS (pH 4.0) containing 1.0 mg·L-1 Cu@PDA, 4.0 g·L-1 Gox and 0.5 mmol·L-1 TMB and the total volume of the solution was made up to 10.0 mL. After standing for 10 min at 40 ℃, it was found that linear relationship between values of absorbance and concentration of glucose was kept in the range of 1.0-30.0 mmol·L-1, with the detection limit (3S/N) of 0.3 mmol·L-1. Samples of human blood were analyzed by the proposed method, giving results of glucose contents in consistency with those reported by the clinical laboratory of the hospital.

       

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