A Two-Mediator System Based on a Nanocomposite of Redox-Active Polymer Poly(thionine) and SWCNT as an Effective Electron Carrier for Eukaryotic Microorganisms in Biosensor Analyzers
Abstract
:1. Introduction
2. Materials and Methods
2.1. Reagents and Equipment
2.2. Cultivation of Microorganism Cells
2.3. Thionine Electropolymerization
2.4. Formation of the Working Electrode
2.5. Registration of Current-Voltage Dependences
2.6. Biosensor Measurements
2.7. Impedance Spectroscopy
2.8. IR Spectroscopy
2.9. Scanning Electron Microscopy (SEM)
2.10. Long-Term Stability of Working Electrodes
2.11. Operational Stability of Working Electrodes
2.12. Determination of BOD by the Standard Dilution Method
3. Results
3.1. Selection of Carbon Nanotubes for the Formation of Conducting Nanocomposite Systems
3.2. Formation of Nanocomposite Materials Based on Redox-Active Biocompatible Polymers
3.3. Electrochemical Properties of the Created Conducting Systems
3.4. Analytical and Metrological Characteristics of BOD Biosensors Based on the Developed Conducting Systems
3.5. Analysis of Water Samples with a Developed Biosensor and a Standard Method for Determining BOD5
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Mediator | Heterogeneous Electron Transfer Rate Constant, (cm·c−1) | Coefficient of Diffusion (cm2/s) | The Rate-Limiting Step of Electron Transfer |
---|---|---|---|
Graphite paste electrode without modification | |||
DCPIP | 0.0018 ± 0.003 | 8.57 × 10−6 | diffusion controlled process |
NR | 0.000921 ± 0.000002 | 6.23 × 10−6 | diffusion controlled process |
Thionine | 0.0026 ± 0.0004 | 5.21 × 10−6 | diffusion controlled process |
Graphene oxide | |||
DCPIP | 0.081 ± 0.003 | - | electron transfer kinetic |
NR | 0.0020 ± 0.0005 | - | electron transfer kinetic |
Thionine | 1.012 ± 0.005 | - | electron transfer kinetic |
SWCNT | |||
DCPIP | 0.39 ± 0.04 | - | electron transfer kinetic |
NR | 0.68 ± 0.01 | - | electron transfer kinetic |
Thionine | 1.228 ± 0.002 | - | electron transfer kinetic |
MWCNT | |||
DCPIP | 0.15 ± 0.04 | - | electron transfer kinetic |
NR | 0.062 ± 0.002 | - | electron transfer kinetic |
Thionine | 0.31 ± 0.02 | - | electron transfer kinetic |
MWCNT –COOH | |||
DCPIP | 0.089 ± 0.002 | - | electron transfer kinetic |
NR | 0.015 ± 0.001 | - | electron transfer kinetic |
Thionine | 0.12 ± 0.02 | - | electron transfer kinetic |
MWCNT –CONH2 | |||
DCPIP | 0.000826 ± 0.00002 | - | electron transfer kinetic |
NR | 0.082 ± 0.005 | - | electron transfer kinetic |
Thionine | 0.00112 ± 0.00002 | - | electron transfer kinetic |
Mediator/Nanomaterials | Heterogeneous Electron Transfer Rate Constant, (cm·c−1) | Reference |
---|---|---|
Thionine/SWCNT | 1.228 ± 0.002 | This work |
Fe[(CN)6]3−/4−/MWCNT | 4.38 | [40] |
Metanil yellow/MWCNT | 11.74 | [41] |
NR/PbO2/α-Al2O3 composite | 35.97 × 10−3 | [42] |
GOD/graphene-chitosan | 2.83 | [43] |
GOD-graphene | 2.68 | [44] |
GOD/PDDAG | 1.59 | [45] |
Conducting System | Rate Constant of Heterogeneous Electron Transfer (k1), cm·s−1 | The Rate Constant (k2) of the Interaction of the Conductive Polymer with the NR, dm3/mol × s | Rate Constant (k3) of Interaction with Microorganisms B. adeninivorans, dm3/(g·s) |
---|---|---|---|
Poly(thionine) | 0.048 ± 0.005 | 1630 ± 80 | 0.00651 ± 0.00003 |
Poly(thionine)- SWCNT | 0.130 ± 0.004 | 5280 ± 90 | 0.07089 ± 0.00002 |
NR [40] | 0.017 ± 0.002 | - | 0.681 ± 0.009 |
Electrode | Rs, Ohm | Cdl, μF | αdl | Rct, kOhm | RW, Ohm | τ, ms | αW |
---|---|---|---|---|---|---|---|
Electrode | 206 | 0.792 | 0.80 | 1900 | - | - | - |
pTN | 254 | 76 | 0.53 | 202 | - | - | - |
pTN/SWCNT | 182 | 301 | 0.68 | 3 | 59 | 5 | 0.37 |
pTN/SWCNT + cells | 159 | 264 | 1 | 0.03 | 132 | 44 | 0.43 |
pTN/SWCNT + cells + NR | 175 | 294 | 0.97 | 0.04 | 39 | 10 | 0.43 |
pTN/SWCNT + cells + substrate | 254 | 259 | 0.98 | 0.06 | 64 | 16 | 0.43 |
pTN/SWCNT + cells + NR + substrate | 247 | 214 | 1 | 0.04 | 118 | 34 | 0.43 |
Characteristic | pTN— B. adeninivorans | pTN—SWCNT— B. adeninivorans | pTN—NR— B. adeninivorans | pTN -SWCNT- NR—B. adeninivorans |
---|---|---|---|---|
Operational stability, % | 6.1 | 4.7 | 3.9 | 3.2 |
Long-term stability, days | 16 | 15 | 18 | 11 |
Duration of a single measurement, min | 6–8 | 6–8 | 6–8 | 6–8 |
Linear range of determined BOD concentrations, mgO2/dm3 | 40–135 | 40–115 | 0.8–320 | 0.4–62 |
Correlation coefficient, R | 0.9705 | 0.9844 | 0.9616 | 0.9729 |
Microorganisms | System | Linear Range of Determined BOD Concentrations, mgO2/dm3 | Reference |
---|---|---|---|
B. adeninivorans | Poly(thionine)-SWCNT-NR | 0.4–62 | This work |
E. coli | NR | 50–1000 | [64] |
Activated sludge | Hexacyanoferrate | 9.8–170 | [65] |
C. violaceum | Hexacyanoferrate | 20–225 | [66] |
D. hanseni | Ferrocene | 25.2–320 | [67] |
D. hanseni | ferrocene–methylene blue | 2.5–7.2 | [68] |
S. cerevisiae | ferricyanide–menadione | 6.6–220 | [69] |
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Medvedeva, A.S.; Dyakova, E.I.; Kuznetsova, L.S.; Mironov, V.G.; Gurkin, G.K.; Rogova, T.V.; Kharkova, A.S.; Melnikov, P.V.; Naumova, A.O.; Butusov, D.N.; et al. A Two-Mediator System Based on a Nanocomposite of Redox-Active Polymer Poly(thionine) and SWCNT as an Effective Electron Carrier for Eukaryotic Microorganisms in Biosensor Analyzers. Polymers 2023, 15, 3335. https://doi.org/10.3390/polym15163335
Medvedeva AS, Dyakova EI, Kuznetsova LS, Mironov VG, Gurkin GK, Rogova TV, Kharkova AS, Melnikov PV, Naumova AO, Butusov DN, et al. A Two-Mediator System Based on a Nanocomposite of Redox-Active Polymer Poly(thionine) and SWCNT as an Effective Electron Carrier for Eukaryotic Microorganisms in Biosensor Analyzers. Polymers. 2023; 15(16):3335. https://doi.org/10.3390/polym15163335
Chicago/Turabian StyleMedvedeva, Anastasia S., Elena I. Dyakova, Lyubov S. Kuznetsova, Vladislav G. Mironov, George K. Gurkin, Tatiana V. Rogova, Anna S. Kharkova, Pavel V. Melnikov, Alina O. Naumova, Denis N. Butusov, and et al. 2023. "A Two-Mediator System Based on a Nanocomposite of Redox-Active Polymer Poly(thionine) and SWCNT as an Effective Electron Carrier for Eukaryotic Microorganisms in Biosensor Analyzers" Polymers 15, no. 16: 3335. https://doi.org/10.3390/polym15163335