Home Physical Sciences A highly sensitive biosensor based on methacrylated graphene oxide-grafted polyaniline for ascorbic acid determination
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A highly sensitive biosensor based on methacrylated graphene oxide-grafted polyaniline for ascorbic acid determination

  • Seyed Morteza Naghib , Farahnaz Behzad , Mehdi Rahmanian , Yasser Zare EMAIL logo and Kyong Yop Rhee EMAIL logo
Published/Copyright: August 30, 2020
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Nanotechnology Reviews
From the journal Nanotechnology Reviews Volume 9 Issue 1

Abstract

Functionalized graphene-based nanocomposites have opened new windows to address some challenges for increasing the sensitivity, accuracy and functionality of biosensors. Polyaniline (PANI) is one of the most potentially promising and technologically important conducting polymers, which brings together the electrical features of metals with intriguing properties of plastics including facile processing and controllable chemical and physical properties. PANI/graphene nanocomposites have attracted intense interest in various fields due to unique physicochemical properties including high conductivity, facile preparation and intriguing redox behavior. In this article, a functionalized graphene-grafted nanostructured PANI nanocomposite was applied for determining the ascorbic acid (AA) level. A significant current response was observed after treating the electrode surface with methacrylated graphene oxide (MeGO)/PANI nanocomposite. The amperometric responses showed a robust linear range of 8–5,000 µM and detection limit of 2 µM (N = 5). Excellent sensor selectivity was demonstrated in the presence of electroactive components interfering species, commonly found in real serum samples. This sensor is a promising candidate for rapid and selective determination of AA.

Keywords: polyaniline; methacrylated graphene oxide; nanocomposite; electrochemical biosensor; ascorbic acid

1 Introduction

Biosensors have attracted much attention due to their unique properties such as simple procedure, easy production, fast response and cost efficiency [1,2,3]. Polyaniline (PANI) is a semi-flexible conducting polymer of the organic semiconductor family [4], which has attracted intensive interest as a result of remarkable features including superior conductivity [5], environmental stability [6], intriguing redox process [7] and inexpensive starting material [8]. In multidisciplinary areas, various applications for PANI have been reported such as biosensors, supercapacitors, biofuel cells, actuators, corrosion protection, membranes, solar cell devices, and rechargeable batteries [4,9,10,11,12]. Tunable properties, good processability (facile synthesis process), affordability, suitable electrochemical and environmental stability, strong bimolecular interactions and intriguing acid/base and doping/dedoping properties have made PANI a promising polymer among inherently conducting polymers [10].

Graphene, a single layer of carbon atoms with sp2 chemical bonds, is the base for all nanoscale carbon materials such as fluorine bucky balls and carbon nanotubes (CNTs) [13,14,15,16,17,18,19,20]. Simple production procedures in comparison with other carbon nanomaterials and several properties, including zero band gap, high conductivity, flexibility, exceptional chemical stability, extremely wide porous structure, high specific surface area, high mobility of charge carriers and cost efficiency, make the graphene a promising nanomaterial for the next era [21]. Graphene-based materials have been extensively used in biosensor applications due to their exceptional electrical, electrochemical and optical characteristics [22,23].

Nanocomposites have attracted much attention due to their exceptional properties [17,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41]. Mechanical performance and electrical conductivity are the main characteristics that are developed with the combination of various materials into a nanocomposite [42,43,44,45,46]. Recently, conductive nanocomposites have captured the great interest in bioelectronics and biosensing fields [47]. Among them, graphene-grafted PANI nanocomposites are valuable owing to their excellent characteristics [48,49,50]. Combination of conducting polymers into a conductive nanostructure enhances the capacity, sensitivity, selectivity and electrical conductivity depending on the preparation methods and morphology. There are π-conjugated electrons in both graphene and PANI. These composites possessed several features including enhanced mechanical strength and excellent electrical conductivity [51]. Also, graphene-grafted PANI nanocomposites are utilized for preparing the electrode substrates. Recently, graphene-grafted PANI nanocomposites with excellent electrochemical characteristics and great conductivity have been utilized for numerous purposes such as biosensors, energy storage tools and electrochemical devices [6].

Ascorbic acid (AA), a reducing agent and successful antioxidant, plays some roles in preventing radical-induced ailments such as tumors and neurodegenerative [14,52]. The deficiency of AA can cause scrubbing, whereas its overdose can lead to stomach cramps and diarrhea [53]. The determination of AA levels is critical for diagnosis of food ingredients. There is a crucial necessity for determining AA level for healthcare and food quality/security due to the healthiness and industrial worth of AA and its low dose in biological and food samples [52].

Electrochemical methods have established rapid and low-cost performance as well as fast response with high selectivity, stability and sensitivity in determining some biomolecules and analytes [54]. Nanostructured composites such as palladium (Pd) nanowire-modified graphene [55], multiwall CNTs dispersed in polyhistidine [56], Fe3O4@gold (Au)-loaded graphene [57] and ZnO nanowire on hierarchical graphene [58] were reported for developing the sensitivity and selectivity of AA. Moreover, other nanocomposites including graphene-grafted PANI [52], graphene-supported platinum (Pt) nanoparticles [59], over-oxidized polypyrrole, PdNPs/Au [60] and 3D graphene foam CuO nanoflowers [61] have been exploited for determining AA.

Our group previously synthesized NFG/AgNPs/PANI for AA biosensing, which was more complex and expensive [52]. Moreover, we synthesized methacrylated graphene oxide (MeGO)/PANI nanocomposite and characterized it by physiochemical and electrochemical tests [6]. In this study, a simpler nanocomposite based on MeGO-grafted PANI is applied as electrochemical biosensor that has several benefits including cost efficiency, high sensitivity and good selectivity over AA determination. The linear range and detection limit of the sensing device are 8–5,000 and 2 µM, respectively. This platform shows the excellent stability over electroactive compounds.

2 Materials and methods

2.1 Chemicals

The potassium ferricyanide (K3Fe(CN)6), potassium permanganate (KMnO4), sulfuric acid (H2SO4), potassium nitrate (KNO3), sodium nitrate (NaNO3) and graphite fine powder (spectroscopic grade, particle size ≤50 µm) were obtained from Merck. Aniline (99%), dimethylformamide, phosphate-buffered saline (PBS), N-hydroxysuccinimide and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride were purchased from Sigma.

2.2 Synthesis of MeGO

Graphite oxide was synthesized via the modified Hummer method [62]. Then, MeGO nanomaterials were prepared based on our previous studies [21,49].

2.3 MeGO-grafted FTO electrode

For preparing the amended electrode substrate, FTO glass plates (8 resistance) with the surface area of 0.25 cm2 were provided and sequential ultrasonic cleaning was performed for 10 min in isopropanol, ethanol, acetone and deionized (DI) water. Under Ar gas flow, the FTO sheets were dried. Then, MeGO suspended in DI was deposited (20, 30 and 40 µL) on the FTO surface by the cast coating method, and it was allowed to dry at 45°C, and the optimum volume was selected (40 µL).

2.4 Electropolymerization of aniline

Electropolymerization of PANI on the electrode surface was established according to our previous study [6]. Briefly, electrodepositing PANI on the MeGO-grafted FTO was initiated with 20 successive cyclic voltammograms (CVs) in a solution consisting of 0.03 M aniline monomer. Then, 0.5 M H2SO4 was applied on the electrode surface.

3 Results and discussion

Transmission electron microscopy (TEM) and field emission scanning electron microscopy (FESEM) tests were applied for investigating the morphology, topography and uniformity of the functionalized MeGO and MeGO/PANI nanocomposite. Figure 1a depicts the TEM image demonstrating a very thin layer of MeGO. The MeGO synthesized in this research was more transparent and uniform in comparison with previous works [63]. Based on previous results, the graphene nanosheets with excellent transparency had flake-like shapes and wrinkles and were more stable upon the exposure to the electron beam.

Figure 1 The morphology and uniformity of the amended electrodes: (a) TEM and (b) FESEM images of the MeGO and pellet/flake-like MeGO/PANI, respectively.
Figure 1

The morphology and uniformity of the amended electrodes: (a) TEM and (b) FESEM images of the MeGO and pellet/flake-like MeGO/PANI, respectively.

Figure 1b shows the microstructure of MeGO/PANI sample by the FESEM analysis. A porous structure was observed after electropolymerization of PANI on the MeGO surface. The forming of the pellet/flake-like microstructure and variations in the topography and morphology of the MeGO substrate corroborated the formation of PANI on the surface (Figure 1b). Clearly, the black regions and transparent edges were ascribed to PANI and MeGO nanosheets in MeGO/PANI nanocomposite, respectively. AA, an important factor for the synthesis and maintenance of collagen in tissue regeneration [64], was tested on our biosensor to assess its performance. CVs of the electrode modified with MeGO/PANI nanocomposites were conducted in the absence and the presence of AA (Figure 2). The results showed the highest catalytic effect for the AA solution.

Figure 2 CVs of (a) MeGO/PANI in 0.02 M PBS without AA and (b) with 10 mM AA at the scan rate of 100 mV s−1.
Figure 2

CVs of (a) MeGO/PANI in 0.02 M PBS without AA and (b) with 10 mM AA at the scan rate of 100 mV s−1.

To obtain the reaction mechanism of AA with the electrode surface, the changes in the oxidation peak of AA in the nanocomposite electrode were examined at different scan rates. The increase of scan rates from 10 to 700 mV s−1 slightly changed the oxidation peak potentials (Figure 3). AA oxidation peak current versus scan rate ½ with higher regression coefficient is linear. This indicates that the reaction mechanisms of AA with the electrode surface follow the diffusion mechanism. Moreover, the peak potential shifts to more positive potential with the increasing scan rate, which is another sign for the diffusion mechanism of AA on the electrode surface. Oxidation peak potential of AA catalysis was selected for chronoamperometry techniques to obtain the linear range of the sensor.

Figure 3 The plots of (a) scan rate and (b) peak current versus scan rate ½.
Figure 3

The plots of (a) scan rate and (b) peak current versus scan rate ½.

Successive aliquots of increasing concentrations of AA were tested on the sensor to obtain amperometric responses of the nanocomposite-modified electrodes. The electrode shows amperometric responses proportional to the AA concentration. The MeGO/PANI electrode demonstrates higher current (less uniform response) along with higher noise compared to MeGO electrode at the same concentrations of AA. This can be attributed to the active edges of graphene, which result in better interactions with AA. The nanocomposite electrode presents greater stability than MeGO electrode.

Figure 4a depicts the current increase in AA level from 8 to 5,000 μM in 0.02 M PBS (pH = 7.4). A linear trend is observed between the peak current and the AA level in Figure 4b (with a correlation of R 2 = 0.99). The detection limit of the amperometric responses was evaluated to be 2 µM (S/N = 5). Therefore, by addition of the AA aliquot (dropwise) to the PBS buffer, the current response (output) of the nanocomposite-based biosensor dramatically promotes to the AA redox reaction linearly with analyte biosensing enviable range. As given in Table 1, the electroanalytic and sensing features of the functionalized MeGO/PANI nanocomposite are meaningfully more than the AA biosensors from the previous reports. Some samples have low detection limit, while others show wide linear sensing range. In comparison with other studies, our sensing device shows very low detection limit and wide linear range. Therefore, this strategy for developing an analytical device could be established as a promising protocol to promote the sensing performance.

Figure 4 Calibration curve and amperometric responses (linear range) of the MeGO/PANI-functionalized biosensor for determining AA and investigating the surface redox reaction. Applied potential was +0.8 V.
Figure 4

Calibration curve and amperometric responses (linear range) of the MeGO/PANI-functionalized biosensor for determining AA and investigating the surface redox reaction. Applied potential was +0.8 V.

Table 1

The comparison of the linear range and detection limit of the present study with others

Electrode materials Detection limit (µM) Linear range (µM) Ref.
PANI/PSS/Gr 5 100–1,000 [65]
NG 2.2 5–1,300 [66]
CoPc–MWCNTs 1 10–2,600 [67]
AGCE/ASOD 2 5–400 [68]
PdNi/C 0.5 10–1,800 [69]
MWCNT/CCE 7.71 15–800 [70]
Pt-Au hybrid 103 103–165 [71]
Chitosan-graphene 50 50–1,200 [72]
OMC/Nafion 20 40–800 [73]
ZnO/RM 1.4 15–240 [74]
MBMOR/P 12.1 20–800 [75]
Pd/CNFs 15 50–4,000 [76]
PMPy/Pd 1,000 50–1,000 [77]
DB71 1 1–2,000 [78]
BPPF6/CPE 8 10–3,000 [79]
PPF/GNS 120 400–6,000 [80]
PdNPs-GO — 20–2,280 [81]
Pt/Au/GCE — 24–384 [71]
NFG/AgNPs (1, 90 s)/PANI 8 10–5,460 [52]
NFG/AgNPs (10, 90 s)/PANI 50 50–11,460 [52]
MeGO/PANI 2 8–5,000 Present work

The selectivity of the MeGO/PANI nanocomposite was evaluated in the presence of some interferences. The current responses of the interfering species were also analyzed at the modified electrode. The selectivity of the sensor was tested in PBS 0.02 M (pH = 7.4) with 10 mM interferences and 5 mM AA. As shown in Figure 5, a significant current response was observed for AA redox reaction, while interferences could not influence the current responses. In spite of the high concentrations of interferences, negligible changes were sensed in the sensing outputs, demonstrating the excellent selectivity of the present platform upon AA determination.

Figure 5 The selectivity of MeGO/PANI sensor in the existence of 10 mM of uric acid and glucose and 5 mM of the analyte in 0.02 M PBS. The applied potential was 0.8 V (versus Ag/AgCl).
Figure 5

The selectivity of MeGO/PANI sensor in the existence of 10 mM of uric acid and glucose and 5 mM of the analyte in 0.02 M PBS. The applied potential was 0.8 V (versus Ag/AgCl).

4 Conclusions

The functionalized MeGO/PANI nanocomposite demonstrated an excellent sensing activity over AA redox reaction. The linear sensing range and the detection limit of the sensing platform were dramatically more than most cases. Electroanalytical and biosensing results illustrated that the combination of MeGO as a 2D nanostructure and PANI as a familiar conducting polymer played a significant role in bioelectrochemical sensing applications. AA was scrutinized as a bioanalyte for verifying the declaration. The time-dependent amperometric output of the MeGO/PANI nanocomposite was noteworthy in an extensive linear range. It is concluded that the present sensor is a talented candidate for quick and careful sensing of AA.

Acknowledgments

This work was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (project number: 2020R1A2B5B02002203).

  1. Conflict of interest: The authors declare no conflict of interest regarding the publication of this paper.

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Received: 2020-06-29
Accepted: 2020-08-05
Published Online: 2020-08-30

© 2020 Seyed Morteza Naghib et al., published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

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https://doi.org/10.1515/ntrev-2020-0061
Keywords for this article
polyaniline; methacrylated graphene oxide; nanocomposite; electrochemical biosensor; ascorbic acid
Creative Commons
BY 4.0

Articles in the same Issue

  1. Research Articles
  2. Generalized locally-exact homogenization theory for evaluation of electric conductivity and resistance of multiphase materials
  3. Enhancing ultra-early strength of sulphoaluminate cement-based materials by incorporating graphene oxide
  4. Characterization of mechanical properties of epoxy/nanohybrid composites by nanoindentation
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  83. Review on the research progress of cement-based and geopolymer materials modified by graphene and graphene oxide
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  106. Theoretical calculation of a TiO2-based photocatalyst in the field of water splitting: A review
  107. Two-photon polymerization nanolithography technology for fabrication of stimulus-responsive micro/nano-structures for biomedical applications
  108. A review of passive methods in microchannel heat sink application through advanced geometric structure and nanofluids: Current advancements and challenges
  109. Stress effect on 3D culturing of MC3T3-E1 cells on microporous bovine bone slices
  110. Progress in magnetic Fe3O4 nanomaterials in magnetic resonance imaging
  111. Synthesis of graphene: Potential carbon precursors and approaches
  112. A comprehensive review of the influences of nanoparticles as a fuel additive in an internal combustion engine (ICE)
  113. Advances in layered double hydroxide-based ternary nanocomposites for photocatalysis of contaminants in water
  114. Analysis of functionally graded carbon nanotube-reinforced composite structures: A review
  115. Application of nanomaterials in ultra-high performance concrete: A review
  116. Therapeutic strategies and potential implications of silver nanoparticles in the management of skin cancer
  117. Advanced nickel nanoparticles technology: From synthesis to applications
  118. Cobalt magnetic nanoparticles as theranostics: Conceivable or forgettable?
  119. Progress in construction of bio-inspired physico-antimicrobial surfaces
  120. From materials to devices using fused deposition modeling: A state-of-art review
  121. A review for modified Li composite anode: Principle, preparation and challenge
  122. Naturally or artificially constructed nanocellulose architectures for epoxy composites: A review
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