基于SERS的过氧化氢检测法及在发酵液中甘油含量监测中的应用  

Hydrogen Peroxide Detection Method Based on SERS and Its Application in Monitoring of Glycerol Content in Fermented Broth

作　　者：张法楠 刘传志[1] 李柏志[2] 李鸿睿 侯玥[1] ZHANG Fanan;LIU Chuanzhi;LI Baizhi;LI Hongrui;HOU Yue(College of Life Science and Technology,Changchun University of Science and Technology,Changchun 130022,China;College of Pharmacy,Jilin University,Changchun 130021,China)

机构地区：[1]长春理工大学生命科学技术学院,长春130022 [2]吉林大学药学院,长春130021

出　　处：《吉林大学学报（理学版）》2022年第6期1465-1472,共8页Journal of Jilin University:Science Edition

基　　金：吉林省科技发展计划项目(批准号:20170204029SF)。

摘　　要：提出一种以纳米金(AuNP)作为增强信号的表面增强Raman光谱(SERS)方法,用于快速、灵敏地定量分析过氧化氢(H_(2)O_(2)),并应用在酵母菌发酵过程中监测甘油含量的变化.实验结果表明:该方法基于发酵液中酶解的甘油与H_(2)O_(2)浓度正相关,当3,3′,5,5′-四甲基联苯胺(TMB)作为底物时,通过过氧化物酶的催化H_(2)O_(2)作用,产生强SERS活性的TMB^(2+)最终产物,通过AuNP增强信号,可灵敏地检测出H_(2)O_(2)含量,按等物质的量比可对甘油含量进行监测;TMB^(2+)在H_(2)O_(2)浓度为2.75~22μmol/L时与SERS信号强度的自然对数呈线性关系,检出限LOD=2.75μmol/L,R^(2)=0.995,平均CV=5.38%,平均回收率为96.05%,该方法具有较高的灵敏度、精密度和准确性,可应用于H_(2)O_(2)浓度检测以及监测发酵过程中甘油含量的变化.We proposed a surface enhanced Raman spectroscopy(SERS)method using nano gold(AuNP)as enhanced signal,which could be used for rapid and sensitive quantitative analysis of hydrogen peroxide(H_(2)O_(2)),and applied it to monitor the change of glycerol content in the process of yeast fermentation.The experimental results show that the method is based on the positive correlation between enzymatically hydrolyzed glycerol and the concentration of hydrogen peroxide.When 3,3′,5,5′-tetramethylbenzidine(TMB)is used as substrate,TMB^(2+)final product with strong SERS activity is produced through the catalytic H_(2)O_(2) action of peroxidase.The H_(2)O_(2) content can be sensitively detected through AuNP enhanced signal,and the glycerol content can be monitored according to the quantity ratio of equivalent substances.The TMB^(2+)is linear relationship with natural logarithm of SERS signal intensity when H_(2)O_(2) concentration is 2.75—22μmol/L,and the detection limit LOD=2.75μmol/L,R^(2)=0.995,average CV=5.38%,average recovery 96.05%.The method has high sensitivity,precision and accuracy,and can be used to detect the concentration of H_(2)O_(2) and monitor the change of glycerol content in the fermentation process.

关 键 词：表面增强Raman光谱 过氧化氢检测 生物传感器 金纳米粒子 

分 类 号：O657.37[理学—分析化学]