研究目的
To develop a hydrogen peroxide-less HRP-based biosensor for the detection of phenolic compounds by utilizing TiO2 to accumulate reactive oxygen species after H2O2 treatment, avoiding the issues associated with dissolved H2O2 in the measuring solution.
研究成果
The study successfully demonstrated a novel enzymatic sensor for phenols using TiO2 pre-activated with H2O2 to generate surface-confined ROS, eliminating the need for dissolved H2O2. This approach improves sensitivity, reduces background noise, and offers a lower detection limit, providing a new direction for peroxidase-based biosensors with potential applications in environmental monitoring.
研究不足
The sensor's response levels out at higher concentrations (>2 μM), limiting its range. Operational stability decreases with higher enzyme loading, and the pre-activation step requires careful handling to avoid enzyme inactivation. The method is specific to TiO2 and may not apply to other materials.
1：Experimental Design and Method Selection:
The study designed an enzymatic biosensor using TiO2 as a host matrix for HRP, pre-activated with H2O2 to generate surface-confined ROS, enabling redox cycling of phenolic analytes without dissolved H2O2. Electrochemical and spectroscopic methods were employed to characterize the system.
2：Electrochemical and spectroscopic methods were employed to characterize the system.
Sample Selection and Data Sources:
2. Sample Selection and Data Sources: Phenolic compounds such as hydroquinone (HQ) and 4-aminophenol (4AP) were used as model analytes. TiO2 (Millennium PC500) and HRP were sourced commercially.
3：List of Experimental Equipment and Materials:
Equipment included PalmSens electrochemical workstation, UV-vis spectrometer (SYNERGYTM MX), EPR spectrometer (Bruker ESP300E), screen-printed electrodes (SPE, ItalSens), and materials like H2O2, Nafion, KCl, HQ, 4AP, TiO2, HRP, and phosphate buffer.
4：Experimental Procedures and Operational Workflow:
TiO2 was calcined and impregnated with HRP. Electrodes were modified by drop-casting TiO2-HRP suspensions, pre-activated in H2O2, washed, and used for amperometric detection of phenols. EPR and UV-vis were used to study ROS formation and enzyme activity.
5：Data Analysis Methods:
Electrochemical data were analyzed using PSTrace software. EPR spectra were simulated with Easyspin toolbox. UV-vis data were used to quantify oxidized HQ. Sensitivity and LOD were calculated from calibration curves.
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