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Pilot study of electrochemical reduction of selected nucleotides and double-stranded DNA at pristine micro-/ultrananocrystalline boron-doped diamond electrodes at very negative potentials

MetadataDetails
Publication Date2024-12-20
JournalElectrochimica Acta
AuthorsMichal Augustín, Vlastimil Vyskočil, Ondrej Szabó, Kateƙina Aubrechtová Dragounová, Rene Pfeifer
AnalysisFull AI Review Included

This pilot study successfully demonstrated the electrochemical reduction of various purine and pyrimidine nucleotides (AMP, ADP, ATP, dGMP, CMP, TMP) and double-stranded DNA (dsDNA) using pristine microcrystalline (B-MCDE) and ultrananocrystalline (B-UNCDE) Boron-Doped Diamond Electrodes (BDDEs) at highly negative potentials.

  • Novel Reduction Signals: Electrochemical reduction of AMP and CMP nucleotides was observed in the far region of the Hydrogen Evolution Reaction (HER), a phenomenon previously unreported for carbonaceous electrodes.
  • Surface Termination Effects: O-terminated BDDEs (hydrophilic) generally facilitated the reduction of all tested analytes (except dsDNA), while H-terminated B-MCDE (hydrophobic) failed to reduce Guanosine 5’-monophosphate (GMP).
  • Protocol Optimization: A proposed protocol involving 30 s incubation and N2 purging significantly enhanced the repeatability of results (typically <5% relative standard deviation) and improved signal magnitude, especially on H-B-MCDE surfaces.
  • HER Interference: Voltammetric determination of analytes in the HER region required very high concentrations (e.g., AMP determination was limited to 2×10-3 M and greater at O-B-MCDE) to suppress hydrogen generation effects.
  • Antifouling Properties: The BDDEs exhibited excellent antifouling characteristics, allowing for repeatable determination of high analyte concentrations with minimal passivation.
  • Future Direction: The study validates BDDEs as promising, non-toxic alternatives to mercury electrodes for DNA electrochemistry, particularly when utilizing hydrophilic O-terminated surfaces.
ParameterValueUnitContext
B-MCDE Film Thickness2.5”mDeposited on Si (111) substrate
B-UNCDE Film Thickness3”mDeposited on Si (111) substrate
B-MCDE Grain Size0.5-1”mMicrometre-sized faceted granular structures
B-UNCDE Grain Size20-70nmUltrananocrystalline nature
B-MCDE Specific Resistivity7.6mΩ cmMeasured via four-point probe
B-UNCDE Specific Resistivity3.7mΩ cmMeasured via four-point probe
B-MCDE Boron Doping Level9.2×1020cm-3Estimated from Raman B1a peak (478 cm-1)
B-UNCDE Boron Doping Level2.4×1021cm-3Estimated from Raman B1a peak (458 cm-1)
Electrode Active Area Diameter4.0mmCircular shape (12.6 mm2)
Optimal LSV Scan Rate1000mV/sLinear Sweep Voltammetry (LSV) experiments
LSV Potential Step2.4mVUsed in voltammetric experiments
Analyte Stock Concentration1×10-2MGMP, dGMP, AMP, ADP, ATP, CMP, TMP
AMP Determination Range (O-B-MCDE)2×10-2 to 2×10-3MVoltammetric determination in presence of HER
Electrolyte pH7.4-0.1 mol/L Phosphate Buffer (PB)

The BDDE films were fabricated using a Linear Antenna Microwave Chemical Vapour Deposition (LA MWCVD) reactor on Si (111) substrates, followed by specific electrochemical activation protocols.

ParameterB-MCDB-UNCDUnitNotes
Substrate Temperature (Ts)600600°CConstant for all experiments
Microwave Power (PMW)66kW-
Gas Mixture Pressure (p)3030Pa-
Deposition Duration (t)3030h-
H2 Flow500500sccm-
TMBT Flow (Carbon/Boron Source)510sccmTrimethyl borate
CO2 Flow11sccmCO2 to H2 concentration 0.2%
CH4 Flow (Crystal Size Reduction)-5sccmMethane added for UNCD growth

The pristine BDDEs were subjected to specific electrochemical procedures to achieve O- or H-termination, followed by a standardized voltammetric protocol.

Termination TypeDeposition Potential (Edep)Deposition Time (tdep)Resulting Surface
O-terminated (Anodic)+3.0 V1 minO-B-MCDE / O-B-UNCDE (Hydrophilic)
H-terminated (Cathodic)-3.0 V1 minH-B-MCDE (Hydrophobic)
  1. Deaeration: Analyte solution (15 mL) purged with N2 for 20 minutes.
  2. Surface Renewal: Electrochemical activation (O- or H-termination) performed immediately before measurement.
  3. Incubation (Preadsorption): Working electrode rinsed (5 s) and placed in the analyte solution for 30 s (tinc=30 s).
  4. Measurement: Linear Sweep Voltammetry (LSV) scan performed (1000 mV/s).

The technology developed in this study, utilizing highly conductive and chemically inert boron-doped diamond electrodes, is relevant to several high-value engineering and commercial sectors:

  • Electroanalysis and Biosensing:
    • DNA/Nucleotide Detection: BDDEs provide a stable platform for monitoring DNA damage, drug-DNA interactions, and nucleotide concentrations, offering a non-toxic alternative to traditional mercury electrodes.
    • High-Sensitivity Sensors: The wide potential window and low background currents characteristic of BDDEs enable the detection of analytes (like AMP and CMP) at extreme negative potentials.
  • Wastewater Treatment (WWT):
    • BDDEs are widely used as anodes for the destruction of persistent organic pollutants via the anodic generation of hydroxyl radicals. The high boron content and low sp2 carbon content (achieved in these films) enhance conductivity and suppress competing reactions (HER/OER), improving WWT efficiency.
  • Electrochemical Device Manufacturing:
    • Robust Electrodes: The mechanical and chemical stability of BDDEs makes them ideal for harsh environments or applications requiring frequent, aggressive electrochemical cleaning/renewal.
  • Material Science and Thin Film Technology:
    • Advanced CVD Processes: The precise control over micro- and ultrananocrystalline film growth (using LA MWCVD and TMBT/CH4 gas mixtures) provides a pathway for manufacturing highly customized diamond materials for specific electrochemical needs.
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