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Indirect electrochemical detection of creatinine in human urine samples using a bare boron-doped diamond electrode

MetadataDetails
Publication Date2025-07-01
JournalBulletin of the Chemical Society of Japan
AuthorsZiping Zhang, Genki Ogata, Yasuaki Einaga
InstitutionsKeio University
AnalysisFull AI Review Included

This research presents a novel, simple, and highly effective non-enzymatic electrochemical method for the indirect detection of creatinine (CRE) in human urine using a bare Boron-Doped Diamond (BDD) electrode.

  • Core Innovation: Successful quantification of CRE in complex human urine matrices without requiring any electrode modification, leveraging the inherent stability and wide potential window of bare BDD.
  • Detection Mechanism: The method relies on a continuous electrochemical redox reaction involving CRE and added sodium nitrite (NaNO2) in a neutral solution (D-PBS, pH 6.4).
  • High Sensitivity: A low limit-of-detection (LOD) of 0.0377 mg/dL (3.3 ”M) was achieved, which is significantly lower than the typical healthy urinary CRE range (4.4 to 18 mM).
  • Clinical Validation: Results showed strong agreement with the conventional spectrophotometric JaffĂ© method, yielding a high correlation coefficient (R) of 0.9964.
  • Robustness: The BDD electrode demonstrated excellent resistance to common urinary interferents (urea, glucose, uric acid, albumin, etc.) after 100-fold sample dilution, with current variations less than 5%.
  • POCT Potential: This strategy offers a simple, low-cost, and rapid analytical technique suitable for Point-of-Care Testing (POCT) and long-term health management.
ParameterValueUnitContext
Electrode MaterialBoron-Doped Diamond (BDD)N/ABare, unmodified working electrode
Boron-Carbon (B/C) Ratio1%Atomic ratio in BDD film
LOD (Limit of Detection)0.0377 (3.3)mg/dL (”M)Calculated in D-PBS (0.5 to 2.5 mg/dL range)
Linear Detection Range0.0377 to 200 (3.3 to 17.68)mg/dL (mM)Effective range for CRE detection
Coefficient of Determination (R2)0.9973N/ALinearity for low-concentration CRE (0.5-2.5 mg/dL)
Correlation with Jaffé (R)0.9964N/AAgreement between BDD and spectrophotometric methods
Interference Variation<5%Variation in DPV current compared to blank (after dilution)
BDD Grain Size2 to 5”mPolycrystalline diamond morphology
BDD sp3 Peak1333cm-1Confirmed by Raman spectroscopy
BDD Bandgap5.45eVIntrinsic diamond property
Boron Activation Energy0.37eVMid-gap state introduction for conductivity

The experimental procedure involves BDD electrode fabrication, a specific electrochemical pre-treatment, and a two-step redox detection protocol using NaNO2 as a mediator.

ParameterValueUnitContext
SubstrateSingle-side silicon (100)N/APolished with diamond powder
Deposition MethodMicrowave Plasma-Assisted CVD (AX6500)N/AUsed for BDD film growth
Carbon SourceAcetoneN/AUsed with Trimethyl borate (TMB)
Carrier GasH2N/AUsed during deposition
Total Pressure115TorrChamber pressure during 4-h reaction
Microwave Power5.00kWPower used during deposition

The detection uses a 3-electrode system (BDD working, Ag/AgCl reference, BDD counter) in D-PBS (pH 6.4) containing 3 mg/dL NaNO2.

  1. Cleaning: BDD and cell cleaned via ultrasonication in ultrapure water (3 times, 5 min each).
  2. Surface Activation (CV): Cyclic Voltammetry (CV) cleaning in 0.1 M H2SO4, ranging from -3.0 to +3.0 V (vs. Ag/AgCl) for 40 cycles at 1 V/s.
  3. H-Termination (CA): Cathodic reduction using Chronoamperometry (CA) in 0.1 M H2SO4 at -3.0 V (vs. Ag/AgCl) for 5 min to obtain a stable H-termination surface.
  4. Oxidation Step (CA): Sample mixture oxidized at +1.5 V (vs. Ag/AgCl) for 30 s. This step generates protons, supporting a localized acidic environment necessary for the diazonium reaction among CRE, Cl-, and NaNO2.
  5. Reduction Step (DPV): Subsequent reduction measured using Differential Pulse Voltammetry (DPV) from 0.3 to -1.0 V. The reduction peak current at -0.15 V (vs. Ag/AgCl) is correlated with CRE concentration.
  6. Sample Preparation: Real urine samples were diluted 100-fold with D-PBS (pH 6.4) before testing to mitigate matrix effects.

The use of highly stable, bare BDD electrodes for robust sensing in complex biological fluids positions this technology for several high-value commercial applications, particularly those requiring simplified, reliable, and portable analytical devices.

  • Point-of-Care Testing (POCT):
    • Development of portable, low-cost electrochemical sensors for rapid, non-invasive monitoring of renal function markers (CRE) in urine outside of centralized laboratories.
    • Suitable for resource-limited settings due to the simplicity of the bare electrode setup (no complex modification layers needed).
  • Clinical Diagnostics and Health Management:
    • Monitoring of chronic kidney disease (CKD) progression and assessing glomerular filtration rate (GFR).
    • Supporting therapy optimization in empirical diseases where renal function is critical.
    • Detection of muscle wasting markers (CRE) in patients with conditions like heart failure.
  • Sports and Anti-Doping Control:
    • Rapid testing of urine dilution levels, often used as an indicator for masking stimulants or illegal drugs.
  • BDD Electrode Manufacturing (Relevant to 6ccvd.com):
    • The demonstrated robustness of BDD against biofouling and its wide potential window confirm its utility in electrochemical sensing products.
    • Applications include high-stability electrodes for complex matrix analysis, water quality monitoring, and advanced electrochemistry where conventional electrodes fail due to fouling or narrow windows.
View Original Abstract

Abstract Electrochemical sensors have been recognized as a promising candidate in point-of-care testing, which is due to their sensitivity, time-saving, low cost, and portability. This work aims to develop a simple and sustainable electrochemical method to measure creatinine (CRE) in real urine. While the present electrochemical sensors for CRE detection rely on the modified electrodes for sensing CRE, our work simply used a bare boron-doped diamond (BDD) electrode without any modification to detect CRE in the urine samples. This method consists of a continuous electrochemical redox reaction of CRE with the addition of NaNO2 in the neutral solution. By using this method, CRE in the range of 0.0377 mg/dL (3.3 ÎŒM) to 200 mg/dL (17.68 mM) exhibits linear performance on the BDD electrode. The results are in good agreement with the JaffĂ© method obtained from the typical spectrophotometric analysis. This strategy has the potential to detect urinary CRE, which could be clinically valuable.