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目的:确定7-硼酸频那醇脂香豆素荧光探针测定·OH的优化条件与实际应用的意义。方法:采用经典Fenton反应,以7-硼酸频哪醇脂香豆素荧光探针为·OH捕捉剂,采用L9(34)正交实验,确定该检测体系的主要影响因素。结果:选用pH7.5的Na2CO3-NaHCO3缓冲溶液,35℃,反应3 h,7-硼酸频那醇脂香豆素荧光探针与·OH产生的荧光强度最强。用H2O2作为标准物质产生的可检测的·OH的最低浓度为10μmol/L,线性响应方程为y=0.0966x+4.6219(R2=0.998),RSD=3.70%。结论:7-硼酸频那醇脂香豆素可用于低浓度·OH的测定,为有色抗氧化物质筛查中·OH测定提供一种新的方法。
OBJECTIVE: To determine the optimal conditions for the determination of · OH by 7-boronic acid pinacol ester coumarin fluorescent probe and its practical significance. Methods: By using classical Fenton reaction, L9 (34) orthogonal experiment with 7-boronic acid pinacol ester-coumarin fluorescent probe as OH scavenger to determine the main influencing factors of this detection system. Results: The fluorescence intensity of the coumarin-7-boronic acid pinacoldehyde coumarin with · OH was the strongest when the pH7.5 Na2CO3-NaHCO3 buffer was used at 35 ℃ for 3 h. The lowest detectable concentration of · OH with H2O2 as standard was 10 μmol / L and the linear response was y = 0.0966x + 4.6219 (R2 = 0.998) with RSD = 3.70%. CONCLUSION: Biocoumarin from 7-boronic acid pinacol can be used for the determination of · OH at low concentration. It provides a new method for the determination of · OH in the screening of colored anti-oxidants.