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    流动注射-分光光度法测定萝卜中过氧化物酶的活性

    FI-Spectrophotometric Determination of Peroxidase Activity of Radish

    • 摘要: 采用流动注射-分光光度法测定萝卜中过氧化物酶的活性。优化的试验条件如下:① 反应温度为60 ℃;② 载流为pH 5.5的0.05 mol·L-1磷酸二氢钠-磷酸氢二钠缓冲溶液;③ 采样体积为40 μL;④ 反应盘管长度为200 cm;⑤ 反应试剂为1.5 mmol·L-1过氧化氢,3.5 mmol·L-1愈创木酚的混合液。2-愈创木酚的表观摩尔吸光率为0.012 5 L·μmol-1·cm-1。辣根过氧化物酶的线性范围为454~7 265 U·L-1,检出限(3s/k)为7 U·L-1。方法的加标回收率在94.2%~107%之间。对500,2 500 U·L-1的辣根过氧化物酶标准溶液连续测定11次,测定值的相对标准偏差小于0.62%。

       

      Abstract: FI-spectrophotometry was applied to the determination of activity of peroxidase in radish. The optimized conditions found were as follows: ① reaction temperature: 60 ℃; ② carrying solution: 0.05 mol·L-1 NaH2PO4-Na2HPO4 buffer solution (pH 5.5); ③ sampling volume: 40 μL; ④ length of reaction coil: 200 cm; ⑤ reaction reagent: mixture of 1.5 mmol·L-1 H2O2 and 3.5 mmol·L-1 2-guaiacol. Apparent molar absorptivity of 2-guaiacol found was 0.012 5 L·μmol-1·cm-1. Linearity range of horse radish peroxidase was found between 454-7 265 U·L-1 with detection limit (3s/k) of 7 U·L-1. Results of test for recovery of this method were found in the range of 94.2%-107%. Precision of the method was tested at the concentration levels of 500, 2 500 U·L-1 of horse radish peroxidase standard solution for 11 determination, values of RSD′s found were less than 0.62%.

       

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