基于氧化型3,3’,5,5’-四甲基联苯胺纳米材料的比色和光热双模式检测谷胱甘肽

Dual-mode detection of glutathione based on the color and photothermal effect of oxidized 3,3’,5,5’ -tetramethylbenzidine nanomaterial

中文关键词:  谷胱甘肽  光热检测  比色检测  普鲁士蓝  3,3’,5,5’-四甲基联苯胺

英文关键词:glutathione  photothermal detection  colorimetric detection  Prussian blue  3,3’,5,5’-tetramethylbenzidine

中文摘要:

      目的 建立基于氧化型3,3’,5,5’-四甲基联苯胺(3,3’,5,5’-tetramethylbenzidine, TMB)的比色和光热双模式谷胱甘肽(glutathione, GSH)检测方法, 用于保健食品和果蔬中谷胱甘肽含量的快速检测。方法 利用普鲁士蓝(Prussian blue, PB)的过氧化物酶活性催化TMB与过氧化氢反应生成蓝绿色且具有光热转化性能的氧化型TMB。而GSH能将氧化型TMB还原为无色的TMB, 使溶液吸光度值和光热转化产生的温升与GSH含量呈线性相关, 实现比色法和光热法双模式定量检测GSH。结果 检测最优pH为3.5, 反应时间为5 min, PB质量浓度为1 mg/mL, 1 mL显色液中30%过氧化氢体积为2.5 μL, TMB质量浓度为15 mg/mL, 光热照射时间为70 s, 照射功率为3.5 W, 最优条件下比色法检出限为14.8 μg/mL, 光热法为1.1 μg/mL, 光热检测灵敏度较比色法提高约13倍, 实际样品加标回收率在84%~119%之间。结论 该方法具有良好的重复性、特异性和准确性, 可用于保健食品、果蔬中GSH的快速检测。

英文摘要:

      Objective To establish a colorimetric and photothermal dual-mode glutathione (GSH) detection method based on oxidized 3,3’,5,5’-tetramethylbenzidine (TMB) for rapid detection of glutathione content in health food, fruits and vegetables.. Methods Prussian blue (PB) was used to catalyze the reaction of TMB and hydrogen peroxide by its peroxidase activity to generate blue-green oxidized TMB with photothermal conversion property. However, GSH could reduce the oxidized TMB to colorless TMB, so that the absorbance value of the solution and the temperature rise generated by photothermal conversion were linearly related to GSH content, thus realizing the dual-mode quantitative detection of GSH by colorimetric method and photothermal method. Results The optimal pH value was 3.5, reaction time was 5 min, PB mass concentration was 1 mg/mL, 30% hydrogen peroxide volume was 2.5 μL in 1 mL chromogenic solution, TMB mass concentration was 15 mg/mL, photothermal irradiation time was 70 s, and irradiation power was 3.5 W. Under the optimal conditions, the limit of detection of colorimetric method was 14.8 μg/mL, and the limit of detection of photothermal method was 1.1 μg/mL, the sensitivity of photothermal detection was about 13 times higher than that of colorimetric method, the actual recoveries of spiked samples were between 84% and 119%. Conclusion This method has good repeatability, specificity and accuracy, and can be used for rapid detection of GSH in health food, fruits and vegetables.