Development of Carbon Cloth/PMG/PAMAM/MWCNTs/ADH Bioanode for Ethanol Biofuel Cell


avatar Ashraf Piryaee 1 , avatar Mahdi Alijanianzadeh 2 , * , avatar Ahmad Molaeirad 1 , avatar Fariba Mashayekhi Mazar 1

Malek-Ashtar University of Technology, Tehran, Iran
Department of Cell and Molecular Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran

how to cite: Piryaee A, Alijanianzadeh M, Molaeirad A, Mashayekhi Mazar F. Development of Carbon Cloth/PMG/PAMAM/MWCNTs/ADH Bioanode for Ethanol Biofuel Cell. J Clin Res Paramed Sci. 2022;11(2):e128811.


More recently, enzyme-based biofuel cells have attracted special attention in different areas. The alcohol dehydrogenase (ADH) due to its cheap and easily available substrate (ethanol) is one of the dehydrogenase enzymes that has been used on researches extensively. This paper explains procurement and performance of bioanode based ADH. In present paper, we used multi-walled carbon nanotubes (MWCNTs) and polyamidoamine dendrimer (PAMAM) on poly methylene green (PMG) modified carbon cloth to fabricate bioanode for an ethanol biofuel cell. The cyclic voltammetry experiments have used to study the performance of the immobilized ADH in presence of β-nicotinamide adenine dinucleotide (NAD+) and ethanol as cofactor and substrate respectively. Also, the kinetic parameter values like kcat, KMS, and k were calculated using analytical equations based on results obtained from cyclic voltammetry. The maximum anodic current of 153.37 µA obtained from the modified bioanode at a concentration of 300 mM ethanol. The kinetic parameter values kcat, KM, and k (rate constant) for enzyme-substrate reaction calculated 1.6 ± 0.1 S-1, 4.4 mM and 1.9 × 105 M-1S-1 respectively. The results showed that the composition of PAMAM and MWCNTs provides a good electronic communication for activity of ADH based on PMG/PAMAM/MWCNTs.

1. Background

Fuel cells are one of the alternative energy sources of the production of electrical energy (1). Biofuel cells (BFC) are a certain type of fuel cells which use biological molecules for the production of electrical energy (2). BFC have advantages including the use of renewable bio-catalysts, the possibility of using a variety of fuels, the ability to operate at low temperatures, mild conditions and physiological pH (3). Among biofuel cells, enzyme-based fuel cells (EBFC) have attracted particular attention (4). Today, various enzymes like alcohol dehydrogenase (ADH) and glucose oxidase and so on have been used to design the enzymatic fuel cells. ADH enzyme reduces NAD+ to NADH during oxidation of ethanol (5). There are many reports on using ADH to design BFC (6-8). In 1997, Palmore et al. used ADH enzyme to design the methanol/O2 biofuel cell (9). In 2008, Sokic-Lazic and Minteer used ADH, aldehyde dehydrogenase (AldDH) and S-acetyl-CoA synthetase with enzymes of the citric acid cycle to oxidize methanol completely (10). In general, EBFC divided into two groups of mediated electron transfer (MET) and direct electron transfer (DET) in term of electron transfer types. In DET, electrons are transferred directly to the electrode surface (11). In MET based EBFC, mediators move the electrons to the electrode surface (12). In ADH-based fuel cell often a mediator used to transfer electrons. Various factors including selection of the appropriate support (electrode materials) and method of enzyme immobilization affect BFC efficiency (13). Carbon cloth is one of the best platforms for effective immobilization of various enzymes. They have a three-dimensional network structure and high thickness as a support for biofuel cell set up (14). One of the key points in studying of NAD+-dependent enzymes is restoration of the NAD+ (15, 16). Because direct oxidation of NADH species in different electrodes done at high potentials (17, 18). The electrocatalysts are commonly used to reduce potential of NADH oxidation (19, 20). Various organic compounds such as methylene green (MG), methylene blue and natural red are used greatly as electrocatalyst (21, 22). Carbon nanotubes (CNTs) is one of the materials that used to recycle the NAD+ on biodevices (23). There are many reports on using CNTs to reduce potential of electrochemical oxidation of NADH (8, 24, 25). Furthermore, CNTs have attracted significant attention to make bioelectrodes due to containing excellent electronic properties, high catalytic activity and biocompatibility. They also cause better enzyme orientation and higher enzyme loading (26-28). There are several methods for the enzyme immobilization (29).Various polymers have been used for enzyme immobilization in EBFC (30, 31). PAMAM dendrimer is one of the polymers that has been widely used for immobilization of enzymes on the electrode surface in recent years (32-34). PAMAM dendrimer is a kind of branched polymers that have different features including large uniformity, narrow molecular weight distribution and highly functionalized terminal surface (35). PAMAM dendrimer due to the presence of end groups keep the enzyme on its surface, also its porous and uniform structure maintains the activity of the enzyme (36). G4 PAMAM dendrimer is the smallest spheroid generation of denderimers with interior spaces (37). Astruc and his colleagues confirmed electrochemical reversibility of redox system terminated dendrimers. They introduced for their phenomenon, electron hopping as proposed mechanism for electron transfer of dendrimer modified electrode (37). The Albery theory is applied to analyze the experimental cyclic voltammetry data in order to extract kinetic parameters of the reaction (38).

2. Methods

2.1. Materials

Alcohol dehydrogenase (ADH: E.C. from Saccharomyces cerevisiae (lyophilized powder, 433 Units mg-1), ncotinamide adenine dinucleotide hydrate (NAD+), polyamidoamine generation 4 dendrimer (PAMAM) and methylene green (MG) and multiwalled carbon nanotubes (MWCNTs) purchased from Sigma-Aldrich (U.S.A). Sodium tetraborate (Na2B4O7), sodium nitrate (NaNO3), Sodium phosphate dibasic (Na2HPO4), Sodium phosphate monobasic monohydrate (NaH2PO4. H2O) and Ethanol were purchased from Merck (Germany). Carbon cloth was purchased from Azar Electrode Company (Iran). All solutions were prepared with deionized water and stored at 4°C when not in use.

2.2. Bioanode Preparation

First, a layer of methylene green was electropolymerized by electrochemically cycling on carbon cloth with area of 1 cm × 1 cm in a solution of 0.4 mmol-1 MG and 100 mmol-1 sodium nitrate and 10 mmol-1 sodium tetraborate in the range of -0.3 V to 1.3 V vs. Ag/AgCl with scan rate of 0.05 V s-1 (39). Then, MG film was washed slowly with double distilled water and dried for 24 hours at room temperature. In the next step, 50 µL of commercial PAMAM solution was dropped on poly methylene green (PMG) modified carbon cloth. After drying PAMAM, 50 µl of MWCNTs was deposited on the electrode surface. Then, modified electrode (carbon cloth/PMG/PAMAM/MWCNTs) was allowed to dry completely. Finally, 50 µL of solution enzyme/NAD+ with concentration of 1 mgml-1 enzyme and 1.9 mM NAD+ was casted on the prepared electrode. The modified carbon cloth by PMG/PAMAM/MWCNTs/ADH was used as a bioanode.

2.3. Electrochemical Measurements

All electrochemical measurements of modified carbon cloth were done in a three-electrode cell chamber (Autolab). In this cell, the modified carbon cloth by PMG/PAMAM/MWCNTs/ADH was used as a working electrode. Chamber cell contained 100 mM phosphate buffer (pH 7.4), 1.9 mM NAD+ and various concentrations of ethanol. All the tests have been done at room temperature.

2.4. Derivation Kinetic Parameters from Cyclic Voltammetry

The following scheme shows the homogeneous system that considered by Albery et al. (40). The reactions occurring in solution are:


At the electrode surface:


In this reaction scheme, S and P are substrate and product. Eox and Ered are the oxidized and reduced forms of the enzyme, and Mox and Mred are the oxidized and reduced forms of the mediator (Figure 1).

The homogeneous system that studied in references (40, 41). Mox and Mred: Oxidized and reduced mediator. Eox and Ered: oxidized and reduced enzyme. S and P: substrate and product. DE and DM: diffusion coefficients of enzyme and mediator.

Albery et al. derived solutions to extract kinetic data from experimental results for different limiting cases (40).

We used the model proposed (case I) by Albery for mediator-enzyme limited kinetics (40).

The amperometric response is given by Equation 1.

Equation 1.I1=nFAmΣDMkeΣ12

In the above equation mΣ, eΣ, k, A, F and n are the concentration of mediator, the enzyme concentration, the rate constant of the reaction between the enzyme and the mediator, the electrode area, faraday constant and number of electrons transferred respectively (38).

Albery et al. also wrote diffusion–reaction equations for enzyme-substrate limited kinetics (cases VI) (40).

Equation 2.IVI=nFA2DMkcateΣmΣSS+KMS12

The enzyme–substrate kinetics can be calculated from plot of the catalytic current against the substrate concentration. The number of electrons transmitted by the MG mediator is n = 2 (38).

3. Results and Discussion

3.1. Cyclic Voltammetry (CV) Characterization

Figure 2 shows the anodic current of the prepared bioanode (carbon cloth/PMG/PAMAM/MWCNTs/ADH) against different concentrations of ethanol in the range of -1 V to +1 V vs. Ag/AgCl in 100 mM phosphate buffer (pH = 7.4), with scan rate of 0.001 V s-1 at 25°C.

Plot of the carbon cloth/PMG/PAMAM/MWCNTs/ADH electrode currents against different concentrations of ethanol in the range of -1 V to +1 V vs. Ag/AgCl in 100 mM phosphate buffer, pH = 7.4, with scan rate of 0.001 V s-1 at 25°C.

Figure 3 shows the cyclic voltammogram of carbon cloth/PMG/PAMAM/MWCNTs/ADH in 100 mM phosphate buffer in absence of NAD+ and ethanol at scan rate of 0.001 V s-1. As it is shown two anodic peaks at -0.17 V and -0.04 V (which corresponds to the performance of electrochemical NAD+ by PMG and MCNTs) and a weak cathodic peak is exist (blue line). A pair of anodic peaks corresponded to the NAD+/NADH redox process is observed in the presence of ethanol. NAD+ is efficiently reduced with higher potential in the presence of ADH. Enhancement in anodic peak current in the presence of ethanol at carbon cloth/PMG/PAMAM/MWCNTs/ADH suggest that ADH shows electrocatalytic effect for ethanol oxidation. By increasing the concentration of ethanol (from 100 mM to 300 mM) current of anodic peak in potential of 0.25 V is increased more than anodic peak in potential of -0.18 V that represents the increase in NADH electrocatalytic oxidation by the ADH (orange line).

Cyclic voltammogram of carbon cloth/PMG/PAMAM/MWCNTs/ADH electrode in 100 mM phosphate buffer (pH = 7.4), between -1.0 V to + 1.0 V vs. Ag/AgCl, at scan rate of 0.001V s-1 and temperature of 25°C, in absence of ethanol (blue) and in presence of ethanol 100 mM (red line), 200 mM (green line) and 300 mM (orange line).

3.2. Kinetic Data

The KMS and kcat are calculated from a plot of the anodic current against ethanol concentration (Figure 2). Currents were taken from the plateau currents in the cyclic voltammograms at 0.25 V and scan rate of 0.001V s-1.

KMS is equal to the substrate concentration at which the current is half of the maximum current. For when the concentration of substrate is very low (s << KMS), anodic current has a linear relationship to the square root of the substrate concentration (38). As a result, Equation 2 is converted to the following equation.

Equation 3.I=nFA 2DMkcatKMSeΣmΣS12

Thus a plot of anodic current against the square root of the substrate concentration (s1/2) shown in Figure 4.

Plot of the carbon cloth/PMG/PAMAM/MWCNTs/ADH electrode currents as a function of the square root on different concentrations of ethanol in the range of -1 V to +1 V vs. Ag/AgCl in 100 mM phosphate buffer, pH = 7.4, with scan rate of 0.001 V s-1 at 25°C.

From plot of anodic current against the square root of the substrate concentration as well as can be calculated the rate constant kcat from current in the saturated region, IMAX, when (s >> KMS).

Equation 4.IMAX=nFA 2DMkcateΣmΣ12

In this study, increasing the concentration of substrate gradually leads to an increase in the amount of anodic current to its maximum.

Table 1 shows the kinetic parameters, kcat, KMS, and k for ADH kinetics mediated by the soluble MG by using analytical equations that have discussed.

Table 1. Kinetic Parameters Obtained from Cyclic Voltammetry Experiments
ParametersEquation 1Equation 2Equation 4
KMS (mM)4.4
kcat (s-1)1.6 ± 0.1
K × 105 (M-1 s-1)1.9 ± 0.1
kcat/KMS × 102 ( M-1 s-1)3.77 ± 0.3

The average concentration of ADH (Γmol cm-2) on the surface of modified carbon cloth calculated from the formula of Q = nFAΓ, where Q, A, F, and n are the charge consumed (C), the electrode area (cm2), F the faraday constant (sAmol-1), and the number of transferred electrons respectively. The immobilized ADH concentration on the modified electrode was estimated approximately 4.152 × 10-6 mol cm-2 (n = 2).

3.3. Effect of Scan Rate

The scan rate expresses rate of the applied potential in cyclic voltammetric experiments. The Randles−Sevcik equation in electrochemically reversible systems shows the linear relationship between anodic current with the square root of the scan rate (Equation 5).

Equation 5.ip=0.446 nFAC0nFvD0RT12

In the above equation ip is the anodic current (A), v is the scan rate (V s-1), n is the number of transferred electrons, A (cm2) is the electrode surface area, Do is the diffusion coefficient of the oxidized species (cm2 s-1), and C0 is the bulk concentration of the analyte (mol cm-3). For reversible processes, it is expected that current to change linearly with increasing scan rates (42). Therefore, the Randles-Sevcik equation and plots of ip versus v1/2 shows whether an analyte is freely diffusing in solution. The effect of different scan rates on oxidation of ethanol onto carbon cloth/PMG/PAMAM/MWCNTs/ADH was investigated by cyclic voltammetry (Figure 5). As the Figure 6 shows, anodic peak current is increased linearly with increasing the scan rate from 0.001 to 0.2 V s-1.

Cyclic voltammograms of carbon cloth/PMG/PAMAM/MWCNTs/ADH electrode in 100 mM phosphate buffer (pH = 7.4), between -0.2 V to 0.8 V vs. Ag/AgCl, scan rate of 0.001 V s-1 to 0.2 V s-1 at 25°C.
The relationship between anodic peak current and scan rates.

3.4. Conclusions

In this study, we used PMG, PAMAM dendrimer and MWCNTs with layer by layer method to do the immobilization of the ADH. The immobilization of MWCNTs on PAMAM film causes facilitate the connection between enzyme and transducer materials, better enzyme orientation and higher enzyme loading. PAMAM dendrimer due to its porous and organizational structure facilitates the transfer of reduced coenzyme to the electrode surface. Therefore, it plays an important role in the reconstruction of the immobilized ADH cycle. Also, the reaction of ADH and PMG has been studied in homogeneous system using cyclic voltammetry and Albery's analytical equations. It is suggested that the combination of simulation and experiment is a more accurate method to study kinetic parameters of enzyme/substrate/mediator systems. However According to the obtained results, MWCNTs and PAMAM create a convenient environment for effective electronic communication between the center redox of enzyme and the electrode surface.


  • 1.

    DeLuca NW, Elabd YA. Polymer electrolyte membranes for the direct methanol fuel cell: A review. J Polym Sci B Polym Phys. 2006;44(16):2201-25.

  • 2.

    Cooney MJ, Svoboda V, Lau C, Martin G, Minteer SD. Enzyme catalysed biofuel cells. Energy & Environmental Science. 2008;1(3):320.

  • 3.

    Bullen RA, Arnot TC, Lakeman JB, Walsh FC. Biofuel cells and their development. Biosens Bioelectron. 2006;21(11):2015-45. [PubMed: 16569499].

  • 4.

    Minteer SD, Liaw BY, Cooney MJ. Enzyme-based biofuel cells. Curr Opin Biotechnol. 2007;18(3):228-34. [PubMed: 17399977].

  • 5.

    Neto AO, Dias RR, Tusi MM, Linardi M, Spinacé EV. Electro-oxidation of methanol and ethanol using PtRu/C, PtSn/C and PtSnRu/C electrocatalysts prepared by an alcohol-reduction process. J Power Sources. 2007;166(1):87-91.

  • 6.

    Umasankar Y, Adhikari BR, Chen A. Effective immobilization of alcohol dehydrogenase on carbon nanoscaffolds for ethanol biofuel cell. Bioelectrochemistry. 2017;118:83-90. [PubMed: 28772201].

  • 7.

    Franco JH, Minteer SD, de Andrade AR. Product Analysis of Operating an Ethanol/O2Biofuel Cell Shows the Synergy between Enzymes within an Enzymatic Cascade. J Electrochem Soc. 2018;165(9):H575-9.

  • 8.

    Villarrubia CW, Garcia SO, Lau C, Atanassov P. Biofuel Cell Anodes Integrating NAD+-Dependent Enzymes and Multiwalled Carbon Nanotube Papers. ECS J Solid State Sci Technol. 2013;2(10):M3156-9.

  • 9.

    Palmore GR, Bertschy H, Bergens SH, Whitesides GM. A methanol/dioxygen biofuel cell that uses NAD+-dependent dehydrogenases as catalysts: application of an electro-enzymatic method to regenerate nicotinamide adenine dinucleotide at low overpotentials. J Electroanal Chem. 1998;443(1):155-61.

  • 10.

    Sokic-Lazic D, Minteer SD. Citric acid cycle biomimic on a carbon electrode. Biosens Bioelectron. 2008;24(4):945-50. [PubMed: 18774285].

  • 11.

    Ghindilis AL, Atanasov P, Wilkins E. Enzyme-catalyzed direct electron transfer: Fundamentals and analytical applications. Electroanalysis. 1997;9(9):661-74.

  • 12.

    Katz E, Willner I. A biofuel cell with electrochemically switchable and tunable power output. J Am Chem Soc. 2003;125(22):6803-13. [PubMed: 12769592].

  • 13.

    Kim J, Jia H, Wang P. Challenges in biocatalysis for enzyme-based biofuel cells. Biotechnol Adv. 2006;24(3):296-308. [PubMed: 16403612].

  • 14.

    Haddad R, Xia W, Guschin DA, Pöller S, Shao M, Vivekananthan J, et al. Carbon Cloth/Carbon Nanotube Electrodes for Biofuel Cells Development. Electroanalysis. 2013;25(1):59-67.

  • 15.

    Kochius S, Magnusson AO, Hollmann F, Schrader J, Holtmann D. Immobilized redox mediators for electrochemical NAD(P)+ regeneration. Appl Microbiol Biotechnol. 2012;93(6):2251-64. [PubMed: 22327354].

  • 16.

    Wu H, Tian C, Song X, Liu C, Yang D, Jiang Z. Methods for the regeneration of nicotinamide coenzymes. Green Chem. 2013;15(7):1773.

  • 17.

    Moiroux J, Elving PJ. Effects of adsorption, electrode material, and operational variables on the oxidation of dihydronicotinamide adenine dinucleotide at carbon electrodes. Anal Chem. 2002;50(8):1056-62.

  • 18.

    Jaegfeldt H. Adsorption and electrochemical oxidation behaviour of NADH at a clean platinum electrode. J Electroanal Chem Interfacial Electrochem. 1980;110(1-3):295-302.

  • 19.

    Lin K, Chen S. Reversible cyclic voltammetry of the NADH/NAD+ redox system on hybrid poly(luminol)/FAD film modified electrodes. J Electroanal Chem. 2006;589(1):52-9.

  • 20.

    Wooten M, Gorski W. Facilitation of NADH electro-oxidation at treated carbon nanotubes. Anal Chem. 2010;82(4):1299-304. [PubMed: 20088562]. [PubMed Central: PMC2821450].

  • 21.

    Malinauskas A, Niaura G, Bloxham S, Ruzgas T, Gorton L. Electropolymerization of Preadsorbed Layers of Some Azine Redox Dyes on Graphite. J Colloid Interface Sci. 2000;230(1):122-7. [PubMed: 10998295].

  • 22.

    Karyakin AA. Electropolymerized Azines: A New Group of Electroactive Polymers. In: Cosnier S, Karyakin A, editors. Electropolymerization: Concepts, Materials and Applications. New Jersey, USA: Wiley‐VCH Verlag GmbH & Co; 2010. p. 93-110.

  • 23.

    Musameh M, Wang J, Merkoci A, Lin Y. Low-potential stable NADH detection at carbon-nanotube-modified glassy carbon electrodes. Electrochem Commun. 2002;4(10):743-6.

  • 24.

    Holzinger M, Le Goff A, Cosnier S. Carbon nanotube/enzyme biofuel cells. Electrochimica Acta. 2012;82:179-90.

  • 25.

    Pelster LN, Meredith MT, Minteer SD. Nicotinamide Adenine Dinucleotide Oxidation Studies at Multiwalled Carbon Nanotube/Polymer Composite Modified Glassy Carbon Electrodes. Electroanalysis. 2012;24(5):1011-8.

  • 26.

    Saifuddin N, Raziah AZ, Junizah AR. Carbon Nanotubes: A Review on Structure and Their Interaction with Proteins. J Chem. 2013;2013:1-18.

  • 27.

    Yan YM, Yehezkeli O, Willner I. Integrated, electrically contacted NAD(P)+-dependent enzyme-carbon nanotube electrodes for biosensors and biofuel cell applications. Chemistry. 2007;13(36):10168-75. [PubMed: 17937376].

  • 28.

    Cosnier S, Holzinger M, Le Goff A. Recent advances in carbon nanotube-based enzymatic fuel cells. Front Bioeng Biotechnol. 2014;2:45. [PubMed: 25386555]. [PubMed Central: PMC4208415].

  • 29.

    Datta S, Christena LR, Rajaram YR. Enzyme immobilization: an overview on techniques and support materials. 3 Biotech. 2013;3(1):1-9. [PubMed: 28324347]. [PubMed Central: PMC3563746].

  • 30.

    Klotzbach TL, Watt M, Ansari Y, Minteer SD. Improving the microenvironment for enzyme immobilization at electrodes by hydrophobically modifying chitosan and Nafion® polymers. J Membr Sci. 2008;311(1-2):81-8.

  • 31.

    Gerard M, Chaubey A, Malhotra BD. Application of conducting polymers to biosensors. Biosens Bioelectron. 2002;17(5):345-59. [PubMed: 11888724].

  • 32.

    Liu Z, Yang Y, Wang H, Liu Y, Shen G, Yu R. A hydrogen peroxide biosensor based on nano-Au/PAMAM dendrimer/cystamine modified gold electrode. Sens Actuators B Chem. 2005;106(1):394-400.

  • 33.

    Cardoso FP, Aquino Neto S, Ciancaglini P, de Andrade AR. The use of PAMAM dendrimers as a platform for laccase immobilization: kinetic characterization of the enzyme. Appl Biochem Biotechnol. 2012;167(7):1854-64. [PubMed: 22639364].

  • 34.

    Deriu D, Favero G, D'Annibale A, Mazzei F. Study of Ferrocene-modified G4 PAMAM Dendrimer for Reagentless Biosensor Develoment. ECS Transactions. 2008;16(11):105-13.

  • 35.

    Tomalia DA, Naylor AM, Goddard WA. Starburst Dendrimers: Molecular-Level Control of Size, Shape, Surface Chemistry, Topology, and Flexibility from Atoms to Macroscopic Matter. Angew Chem, Int Ed Engl. 1990;29(2):138-75.

  • 36.

    Fenga PG, Cardoso FP, Aquino Neto S, De Andrade AR. Multiwalled carbon nanotubes to improve ethanol/air biofuel cells. Electrochimica Acta. 2013;106:109-13.

  • 37.

    Astruc D, Boisselier E, Ornelas C. Dendrimers designed for functions: from physical, photophysical, and supramolecular properties to applications in sensing, catalysis, molecular electronics, photonics, and nanomedicine. Chem Rev. 2010;110(4):1857-959. [PubMed: 20356105].

  • 38.

    Flexer V, Ielmini MV, Calvo EJ, Bartlett PN. Extracting kinetic parameters for homogeneous [Os(bpy)2ClPyCOOH]+ mediated enzyme reactions from cyclic voltammetry and simulations. Bioelectrochemistry. 2008;74(1):201-9. [PubMed: 18824418]. [PubMed Central: PMC2742741].

  • 39.

    Dong-mei Z, Hui-Qun F, Hong-yuan C, Huang-xian J, Yun W. The electrochemical polymerization of methylene green and its electrocatalysis for the oxidation of NADH. Analytica Chimica Acta. 1996;329(1-2):41-8.

  • 40.

    Albery W, Bartlett PN, Driscoll BJ, Lennox R. Amperometric enzyme electrodes. J Electroanal Chem. 1992;323(1-2):77-102.

  • 41.

    Bartlett PN, Pratt KFE. A study of the kinetics of the reaction between ferrocene monocarboxylic acid and glucose oxidase using the rotating-disc electrode. J Electroanal Chem. 1995;397(1-2):53-60.

  • 42.

    Elgrishi N, Rountree KJ, McCarthy BD, Rountree ES, Eisenhart TT, Dempsey JL. A Practical Beginner’s Guide to Cyclic Voltammetry. J Chem Educ. 2017;95(2):197-206.

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