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