Non-Enzymatic Selective Detection of Histamine in Fishery Product Samples on Boron-Doped Diamond Electrodes
At a Glance
Section titled âAt a Glanceâ| Metadata | Details |
|---|---|
| Publication Date | 2025-07-29 |
| Journal | Biosensors |
| Authors | Hiroshi Aoki, Risa Miyazaki, Yasuaki Einaga |
| Institutions | Keio University, National Institute of Advanced Industrial Science and Technology |
| Analysis | Full AI Review Included |
Technical Analysis and Documentation: Non-Enzymatic Histamine Sensing via O-BDD Electrodes
Section titled âTechnical Analysis and Documentation: Non-Enzymatic Histamine Sensing via O-BDD ElectrodesâExecutive Summary
Section titled âExecutive SummaryâThis research successfully demonstrates a novel, non-enzymatic method for the selective and rapid detection of histamine in complex food matrices (fishery products) using Boron-Doped Diamond (BDD) electrodes.
- Non-Enzymatic Advantage: The method bypasses the instability and long processing times associated with traditional enzyme-based histamine sensors, offering a robust, reusable alternative.
- Material Selection: The wide potential window and low background current of MPCVD-grown BDD electrodes were critical for observing the high oxidation potential of histamine.
- Enhanced Selectivity: Selectivity over the primary interferent, histidine, was achieved by combining an oxygen-terminated BDD (O-BDD) surface with an optimized alkaline solution (pH 8.4).
- Electrostatic Mechanism: The negatively charged O-BDD surface selectively attracts the positively charged histamine ions while repelling the negatively charged histidine ions, shifting their respective oxidation potentials.
- High Performance: The sensor exhibited a highly linear response (R2 = 0.968) in fish extracts across the critical food safety range of 0-150 ppm.
- Practical Application: The method achieved a detection limit of 20.9 ppm, confirming its suitability for rapid, on-site quantification of histamine in real-world food quality control.
Technical Specifications
Section titled âTechnical SpecificationsâThe following hard data points were extracted from the analysis of the BDD electrode fabrication and sensor performance:
| Parameter | Value | Unit | Context |
|---|---|---|---|
| Electrode Material | Boron-Doped Diamond (BDD) | N/A | Grown via MPCVD on Si(100) |
| Boron Doping Level | 1 | % | B/C atomic ratio (from trimethyl borate source) |
| BDD Film Thickness | 40 | ”m | Result of 10-hour deposition at 5 kW |
| Working Electrode Area | 0.28 | cm2 | Circular geometry (0.6 cm diameter) |
| Surface Termination | Oxygen-Terminated (O-BDD) | N/A | Achieved via anodic oxidation (+3.0 V) |
| Optimized Measurement pH | 8.4 | N/A | Selected to maximize electrostatic selectivity |
| Electrochemical Method | Linear Sweep Voltammetry (LSV) | N/A | Potential scanned 0 to +2.0 V vs Ag/AgCl |
| LSV Scan Rate | 0.1 | V s-1 | Standard measurement condition |
| Histamine Oxidation Potential | ~+1.05 | V vs Ag/AgCl | Potential used for selective current measurement |
| Linear Response Range | 0-150 | ppm | Histamine concentration in fish extract |
| Limit of Detection (LOD) | 20.9 | ppm | Calculated using S/N = 3.0 |
| Correlation Coefficient | 0.968 | R2 | Linearity of the calibration curve |
Key Methodologies
Section titled âKey MethodologiesâThe BDD electrodes were prepared and activated using precise MPCVD growth and electrochemical post-processing techniques:
- BDD Film Growth: Films were deposited on Si(100) wafers using a Microwave Plasma Assisted Chemical Vapor Deposition (MPCVD) system.
- Precursor Selection: Acetone served as the carbon source, and trimethyl borate was used as the boron source, maintaining a B/C atomic ratio of 1%.
- Physical Parameters: Deposition was conducted for 10 hours at a microwave power of 5 kW, yielding a BDD film thickness of approximately 40 ”m.
- Electrode Configuration: A standard three-electrode cell was used, featuring the BDD film as the working electrode (0.28 cm2), a Pt auxiliary electrode, and an Ag/AgCl reference electrode.
- Surface Activation (O-BDD Termination): The BDD electrodes were electrochemically cleaned and activated via Chronoamperometry (CA) in 1Ă PBS:
- Cathodic Reduction (H-BDD): -3.0 V applied for 5 minutes.
- Anodic Oxidation (O-BDD): +3.0 V applied for 5 minutes, resulting in the necessary oxygen-terminated surface.
- Electrochemical Measurement: Linear Sweep Voltammetry (LSV) was performed by scanning the potential from 0 to +2.0 V at a scan rate of 0.1 V s-1.
6CCVD Solutions & Capabilities
Section titled â6CCVD Solutions & CapabilitiesâThe successful development of this robust, non-enzymatic histamine sensor relies entirely on the high-quality, customizable Boron-Doped Diamond material, a core specialty of 6CCVD. We are uniquely positioned to supply the materials and engineering support necessary to replicate, scale, and advance this research.
Applicable Materials
Section titled âApplicable MaterialsâTo replicate or extend this high-performance electrochemical sensor, 6CCVD recommends the following materials:
- Heavy Boron-Doped PCD (Polycrystalline Diamond): The research required a B/C ratio of 1% (10,000 ppm). 6CCVD routinely manufactures highly conductive BDD films, ensuring the wide potential window and low background current essential for high-potential oxidation reactions like histamine sensing.
- Custom Thickness BDD: The paper utilized a 40 ”m thick film. We offer BDD films in custom thicknesses ranging from 0.1 ”m up to 500 ”m, allowing researchers to optimize conductivity and mechanical stability for specific sensor designs.
Customization Potential
Section titled âCustomization Potentialâ6CCVDâs advanced fabrication capabilities directly address the specific dimensional and integration needs of electrochemical sensor development:
| Research Requirement | 6CCVD Customization Capability | Value Proposition |
|---|---|---|
| Precise Electrode Geometry | Custom Laser Cutting: We provide precision laser cutting services to achieve exact working electrode areas (e.g., 0.28 cm2) and custom shapes required for integration into microfluidic or flow cell systems. | Ensures high reproducibility and integration readiness. |
| Substrate Compatibility | Flexible Substrate Growth: While the paper used Si(100), 6CCVD can grow BDD films on various substrates (Si, Mo, etc.) to meet specific thermal or mechanical requirements of the final device. | Simplifies device integration and manufacturing scale-up. |
| Surface Termination Control | Engineering Consultation: Although the paper used electrochemical oxidation, 6CCVDâs in-house expertise includes optimizing surface termination (H-terminated, O-terminated) via plasma or chemical treatments to ensure the desired electrostatic properties are achieved consistently and at scale. | Guarantees optimal material performance for selective sensing. |
| Scalability | Large Format BDD: We offer PCD/BDD wafers up to 125mm in diameter, enabling the transition from laboratory-scale prototypes (0.6 cm diameter) to high-volume manufacturing of sensor arrays. | Facilitates commercialization and mass production. |
Engineering Support
Section titled âEngineering Supportâ6CCVDâs in-house PhD team specializes in electrochemical diamond applications. We offer comprehensive support to engineers and scientists working on similar non-enzymatic biosensing and electroanalytical projects:
- Material Optimization: Assistance in selecting the optimal BDD doping level, film thickness, and surface roughness (polishing Ra < 5nm for PCD) to maximize sensor sensitivity and stability.
- Process Integration: Consultation on integrating BDD electrodes into complex systems, including advice on metalization schemes (e.g., Au, Pt, Ti, W) for reliable electrical contacts, which we offer as an internal capability.
- Global Logistics: We ensure reliable global shipping (DDU default, DDP available) of sensitive diamond materials, supporting research teams worldwide.
For custom specifications or material consultation, visit 6ccvd.com or contact our engineering team directly.
View Original Abstract
Histamine sensing that uses enzymatic reactions is the most common form of testing due to its selectivity for histamine. However, enzymes are difficult to store for long periods of time, and the inactivation of enzymes decreases the reliability of the results. In this study, we developed a novel, quick, and easily operated histamine sensing technique that takes advantage of the histamine redox reaction and does not require enzyme-based processes. Because the redox potential of histamine is relatively high, we used a boron-doped diamond (BDD) electrode that has a wide potential window. At pH 8.4, which is between the acidity constant of histamine and the isoelectric point of histidine, it was found that an oxygen-terminated BDD surface successfully detected histamine, both selectively and exclusively. Measurements of the sensorâs responses to extracts from fish meat samples that contained histamine at various concentrations revealed that the sensor responds linearly to the histamine concentration, thus allowing it to be used as a calibration curve. The sensor was used to measure histamine in another fish meat sample treated as an unknown sample, and the response was fitted to the calibration curve to perform an inverse estimation. When estimated in this way, the histamine concentration matched the certified value within the range of error. A more detailed examination showed that the sensor response was little affected by the histidine concentration in the sample. The detection limit was 20.9 ppm, and the linear response range was 0-150 ppm. This confirms that this sensing method can be used to measure standard histamine concentrations.