吉林大学学报(理学版) ›› 2022, Vol. 60 ›› Issue (6): 1465-1472.

• • 上一篇    

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

张法楠1,  刘传志1,  李柏志2,  李鸿睿2,  侯玥1   

  1. 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
  • 收稿日期:2021-12-28 出版日期:2022-11-26 发布日期:2022-11-26
  • 通讯作者: 侯玥 E-mail:hy46155@126.com

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

ZHANG Fanan1,  LIU Chuanzhi1,   LI Baizhi2,   LI Hongrui2,   HOU Yue1   

  1. 1. College of Life Science and Technology,  Changchun University of Science and Technology,  Changchun 130022,  China;   2. College of Pharmacy,  Jilin University,  Changchun 130021,  China
  • Received:2021-12-28 Online:2022-11-26 Published:2022-11-26

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(H2O2), 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H2O2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H2O2作用, 产生强SERS活性的TMB2+最终产物, 通过AuNP增强信号, 可灵敏地检测出H2O2含量, 按等物质的量比可对甘油含量进行监测;  TMB2+H2O2浓度为2.75~22 μmol/L时与SERS信号强度的自然对数呈线性关系, 检出限LOD=2.75 μmol/L, R2=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H2O2浓度检测以及监测发酵过程中甘油含量的变化. 

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

Abstract: 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 (H2O2),  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,  TMB2+ final product with strong SERS activity is produced through the catalytic H2O2 action of peroxidase. The H2O2 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 TMB2+ is  linear relationship with natural logarithm of SERS signal  intensity when  H2O2 concentration is 2.75—22 μmol/L,  and the detection limit LOD=2.75 μmol/L,  R2 =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 H2O2 and monitor the change of glycerol content in the fermentation process.

Key words: surface enhanced Raman spectroscopy (SERS),  , hydrogen peroxide detection,  , biosensor,  , gold nanoparticles

中图分类号: 

  • O657.37