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1 DOI: 10.1002/elan.201800656
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An Enzyme-free H2O2 Sensor Based
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on Poly(2-Aminophenylbenzimidazole)/Gold Nanoparticles
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Coated Pencil Graphite Electrode
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10 Mine Şen Teker,[a] Erhan Karaca,[a] Nuran Özçiçek Pekmez,*[a] Uğur Tamer,[b] and Kadir Pekmez[a]
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13 Abstract: A poly(2-aminophenylbenzimidazole)/gold H2O2. The analytical performance was evaluated in a
14 nanoparticles (P2AB/AuNPs) coated disposable pencil 100 mM phosphate buffer solution at pH 6.5 by amperom-
15 graphite electrode (PGE) was fabricated as an enzyme- etry. The steady state current vs. H2O2 concentration is
16 free sensor for the H2O2 determination. P2AB/AuNPs and linear in the range of 0.06 to 100 mM (R2 = 0.992) with a
17 P2AB were successfully synthesized electrochemically on limit of detection 3.67 3 105 M at 0.8 V vs. SCE and no
18 PGE in acetonitrile for the first time. The coatings were interference is caused by ascorbic acid, dopamine, uric
19 characterized by scanning electron microscopy, X-ray acid, and glucose. The examination for the sensitive
20 diffraction spectroscopy, Energy-dispersive X-ray spectro- determination of H2O2 was conducted in commercially
21 scopy, Surface-enhanced Raman spectroscopy, and UV- available hair oxidant solution. The results demonstrate
22 Vis spectroscopy. AuNPs interacted with P2AB as carrier that P2AB/AuNPs/PGE has potential applications as a
23 enhances the electrocatalytic activity towards reduction of sensing material for quantitative determination of H2O2.
24 Keywords: Electrochemical sensor · Poly(2-aminophenylbenzimidazole) · Gold nanoparticles · Pencil graphite electrode · Hydrogen
25 peroxide
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28 ucts and hair dyes. Due to its extensive applications, the
1 Introduction
29 electrochemical sensing of H2O2 has obtained great
30 Conjugated polymers have arisen as promising materials attention in recent years [7]. For example, a Pt nano-
31 for the design of electronic devices the significant particle modified Pencil Graphite Electrode (PGE) was
32 flexibility in their chemical structures in preparation, high prepared for non-enzymatic determination of H2O2 in a
33 surface area, high electrical conductivity, chemical selec- hair dye oxidation cream by Flow Injection Analysis
34 tivity, excellent biocompatibility and their redox charac- system [7c]. In the study by Benvidi et al., a voltammetric
35 teristics [1]. The electrochemical polymerization has many sensor based on a carbon paste electrode with CuFe2O4
36 advantages, including control of their morphology, good nanoparticle was designed for the electrocatalytic reduc-
37 reproducibility, and stability of the polymer film on tion of H2O2 and used for detection of H2O2 in milk, green
38 electrode surfaces based on the applied polymerization tea, hair dye cream and mouthwash solution samples using
39 parameters [2]. Benzimidazoles are a class of heterocyclic, DPV technique [7d]. In another study, an amperometric
40 aromatic compounds which share a fundamental structural sensor was prepared based on gold nanoparticle-deco-
41 characteristic of six-membered benzene fused to five- rated copper cross-linked pectin (CuCP-AuNPs) for H2O2
42 membered imidazole. Benzimidazole has great application determination in the commercially available clinical lens
43 areas such as antitumor, antibacterial agents and corrosion cleaning solutions. [7b].
44 inhibition. It is also of high interest due to two nitrogens Fabricating electrodes with metal nanoparticles for
45 of the amine hydrogen as the donor and the imino determination H2O2 sensors have arisen considerable
46 nitrogen as the acceptor in fuel cell studies [3]. It was
47 proved that electropolymerization of benzimidazole and
48 substituted benzimidazoles produced electroactive poly- [a] M. Ş. Teker, E. Karaca, N. . Pekmez, K. Pekmez
49 mers with increasing the conjugation and conductivity of Hacettepe University
50 the macromolecule [4]. Polybenzimidazole (PB) is an Department of Chemistry
51 aromatic polymer with exceptional chemical resistance, 06800 Ankara, Turkey
52 excellent mechanical strength, and thermal stability [5]. E-mail: npekmez@hacettepe.edu.tr
[b] U. Tamer
53 When doped with acid, it behaves as a proton conductor
Gazi University
54 [6]. Faculty of Pharmacy
55 Hydrogen peroxide (H2O2) detection has practical Department of Analytical Chemistry
56 importance in many fields such as industrial, environ- 06330 Ankara, Turkey
57 mental, waste water treatment, textile, and industries Supporting information for this article is available on the
58 household products, disinfectants, tooth whitening prod- WWW under https://doi.org/10.1002/elan.201800656

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1 attention in recent studies due to their large surface-to- working electrode for scanning electron microscopy
2 volume ratio high electrical conductivity and excellent (SEM), Fourier transform infrared spectroscopy (FTIR),
3 surface reaction activity [8]. Especially gold nanoparticles X-ray power diffraction (XRD) and surface enhanced
4 (AuNPs) could provide a friendly microenvironment and Raman (SERS) measurements. ITO glass electrode (5–
5 act as the conducting tunnel to achieve the direct electron 15 W Delta technologies) was also used as a working
6 transfer between substances and electrode surfaces [9]. electrode for UV-Vis studies. CHI 6011D (CH Instru-
7 AuNPs could be synthesized by reducing aqueous HAuCl4 ments) and Emstat3 + were used in electrochemical
8 solution with the addition of various reducing agents and measurements. Electrochemical impedance spectroscopy
9 stabilizing reagents under different parameters and con- (EIS) technique was also used for electrochemical charac-
10 ditions [10]. The literature reported some AuNPs based terization of the electrode surface. The surface morphol-
11 non-enzymatic H2O2 sensors [11]. The studies continue for ogy and elemental mapping of all samples were charac-
12 the improvement of non-enzymatic H2O2 sensor. terized using NOVA NANOSEM 650/FEI (USA) and
13 There has been no report yet related to polymerization AMETEK-EDAX (USA). XRD pattern was recorded by
14 of 2-aminophenyl benzimidazole and also modification PANalytical/EMPYREAN (USA) with Cu Ka radiation
15 with AuNPs in non-aqueous medium without adding operating at 45 kV and 40 mA. SERS measurements were
16 reducing or stabilizing agents for use as an H2O2 sensor. performed using a DeltaNu Examiner Raman microscope
17 In the present study, gold nanoparticles functionalized 2 (Laramie, WY, USA) with a 785-nm laser source. FTIR
18 aminophenylbenzimidazole was electropolymerized in absorbance spectra were conducted on Thermo Scientific
19 acetonitrile on disposable pencil graphite electrode, and Nicolet iS10 (USA). UV-Vis absorption spectra were
20 poly(2-aminophenylbenzimidazole)/gold nanoparticles collected using a Perkin Elmer Lambda UV-Vis-NIR 750
21 (P2AB/AuNPs) coated electrode was examined for am- spectrophotometer over the 400–800 nm range. The dry
22 perometric analysis of H2O2 for the first time. conductivity values of the polymer were measured using
23 the four-probe measuring technique at room temperature.
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25 2 Experimental
3 Results and Discussion
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2.1 Materials
27 3.1 Preparation and Characterization of P2AB and
28 2-(2-Aminophenyl)-1H-benzimidazole 97 % (2AB), Hy- P2AB/AuNPs Films
29 drogen tetrachloroaurate (HAuCl4), tetrabutylammo-
3.1.1 Preparation of P2AB and P2AB/AuNPs on Pencil
30 niumperchlorate (TBAP), acetonitrile (ACN), sodium
Graphite Electrodes
31 phosphate monobasic dihydrate (NaH2PO4 · 2H2O) so-
32 dium phosphate dibasic dihydrate (Na2HPO4 · 2H2O) and Poly(2-aminophenylbenzimidazole) (P2AB) films with
33 acetic acid (CH3COOH) were obtained from Sigma- and without gold nanoparticles (AuNPs) were deposited
34 Aldrich (Taufkirchen, Germany). Hydrogen peroxide on pencil graphite electrode (PGE) by applying cyclic
35 35 % (Merck/Germany) was freshly prepared before every voltammetry, scanning between 1.0 and 1.4 V vs. Ag j
36 use. Perchloric acid (HClO4) from Analar was used. AgCl at a scan rate of 100 mV s1. The electropolymeriza-
37 Phosphate buffered saline (PBS, 100 mM) was prepared tion of 2-aminophenylbenzimidazole (2AB) was carried
38 from NaH2PO4 · 2H2O, Na2HPO4 · 2H2O and pH value was out in an electrochemical cell containing acetonitrile
39 adjusted to 6.5 by using CH3COOH solution. Commercial solution of 100 mM TBAP, 100 mM 2AB and 75 mM
40 H2O2 hair oxidant (URGO, 3 %) was purchased from a HClO4. 2AB is oxidized after 0.75 V and recorded a broad
41 local market for real sample analysis. All chemicals were peak of about 0.9 V as observed in the first cycle
42 used as received. Double distilled water used throughout (Figure 1A (a)). The formation and growth of the P2AB
43 the study. can easily be seen in Figure 1A (a). The peak intensity
44 due to the oxidation of 2AB gradually decreases, and the
45 broad peaks between 0.2 V and 0.7 V due to the
2.2 Methods
46 oxidation and reduction of the film increase in intensity as
47 Electrochemical measurements were performed with a the film grows. When the 2AB monomer is electro-
48 one compartment glass cell using three electrodes by using polymerized in the presence of HAuCI4 (10.0 mM) (Fig-
49 cyclic voltammetry (CV), amperometric i-t techniques and ure 1A (b)), the small peak of the oxidation of the
50 electrochemical impedance techniques. The working elec- monomer in the first cycle is distinguished, and this peak
51 trode was a disposable graphite electrode (TOMBO 2B disappears slightly during subsequent cycles. The anodic
52 with a diameter of 0.9 mm). All leads were used as and cathodic peak characteristics of the polymer are the
53 received. The auxiliary electrodes were Pt wire with a same that of obtained without HAuCI4 but the peak
54 compartment for non-aqueos solutions and bare Pt wire intensities are higher, and the peak-to-peak separation
55 for aqueous solutions. The electrochemical studies were between 0.7 V and 0.2 V decreases. These findings
56 referenced to Ag/AgCl (3.0 M KCl) electrode and satu- indicate that the polymer formation occurs much faster in
57 rated calomel electrode (SCE) for non-aqueous, aqueous the presence of HAuCI4 and more polymer can be
58 solutions, respectively. Pencil graphite was used as the deposited on the PGE. Electrochemical behavior was

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Fig. 1. (A) Electrochemical deposition of (a) P2AB and (b) P2AB/AuNPs films by scanning the potential between 1.0 and 1.4 V vs.
17 Ag/AgCl on PGE (inset: molecular structure of 2AB). (B) Potential cycling curves recorded at 100 mV s1 in PBS pH 6.5 (100 mM)
18 solution for the coatings (a) P2AB, (b) P2AB/AuNPs, and (c) bare pencil graphite electrode, v = 100 mV s1.
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21 investigated for each coated electrode by cyclic voltamme- electrode. The comparison of the spectra in Figure S1A
22 try, scanning between 1.0 V and 1.0 V vs. SCE at a scan (b) to B (b) shows that the absorption maxima of AuNPs
23 rate of 100 mV s1 in an aqueous phosphate buffer shifts from 535 nm to 555 nm due to electropolymerization
24 solution (PBS) of pH 6.5. The redox peak currents of of 2AB monomers. Therefore, the peaks at 475 nm and
25 P2AB obtained with a HAuCI4 increase when compared 555 nm in Figure S1B (b) may belong to P2AB moieties
26 to those of bare and P2AB coated electrodes (Figure 1B without and with AuNPs, respectively.
27 (b)). It can be interpreted that AuNPs facilitates the XRD pattern (Figure S2) was used to further con-
28 electron transfer process, which means that more polymer firmation for the crystal plane of the prepared P2AB/
29 deposits on the PGE. AuNPs film on PGE. In Figure S2, P2AB/AuNPs film
30 characteristic four different diffraction peaks (2q) at 37.88,
31 42.88, 64.98 and 77.88 are associated with the cubic phase
3.1.2 Spectroscopic Characterization
32 (111, 200, 220, 311) of Au nanoparticles [13]. In addition,
33 UV-Vis measurements of the polymerization solutions the diffraction peaks (2q) around 42.88, 54.88 and 77.88
34 (100 mM TBAP, 100 mM 2AB and 75.0 mM HClO4 in are also corresponding with graphite (101, 004, 103) [14].
35 acetonitrile) without and with HAuCI4 (10.0 mM) were The obtained XRD pattern is similar to the previously
36 performed (Figure S1A (a) and (b)). In the spectrum of reported XRD spectra of gold nanoparticles (JCPDS, card
37 the polymerization solution without HAuCI4, an absorb- no. 04-0784). According to the results, Au nanoparticles
38 ance peak appears at 410 nm (Figure S1A (a)), which is and P2AB polymer coexist on the PGE surface.
39 related to p-p electronic interactions. On the other hand, The FTIR spectroscopy was used to characterize
40 when 10.0 mM HAuCI4 is added to the polymerization P2AB, and P2AB/AuNPs coated PGEs. The absorbance
41 solution, light yellow color immediately turns dark violet, peaks of P2AB/AuNPs are almost similar to those of
42 and an absorption peak occurs at 535 nm, disappearing P2AB homopolymer (Figure 2A), but it is clear that these
43 the peak at 410 nm (Figure S1A (a)). It is well-known that bands shift longer wavenumbers. The marked shift may be
44 the peak at 535 nm is the characteristic surface plasmon due to interaction of the P2AB with AuNPs. In the
45 resonance band of metal nanoparticles [12]. This is the spectrum of P2AB/AuNPs film, a broad band at 3312 cm1
46 sign of synthesis of the AuNPs. Accordingly, HAuCI4 is is corresponding to NH stretching bond [15]. The band
47 reduced to AuNPs in the presence of partially protonated at 2900 cm1 is assigned vibrations of aromatic CH in the
48 2AB monomer in acetonitrile solution. In addition, P2AB polymer chains [16]. The C=N and C=C stretching
49 polymers were deposited on the ITO electrode from the vibrations of the benzimidazole moiety appear as a broad
50 polymerization solutions without and with HAuCI4 by peak around 1600 cm1 [17]. Also, the peak at 1421 cm1 is
51 cyclic voltammetry, and their spectra were taken for ascribed to the ring vibration of the ortho substituted
52 comparison (Figure S1B). The pale yellow colored P2AB phenyl group [16]. The CN stretching and NH bending
53 film gives a broad absorption band at 475 nm, while dark vibrations of the benzimidazole moiety appear around
54 yellow colored P2AB/AuNPs film has two absorption 1319 and 1361 cm1 [16]. At the peak at 1245 cm1 is
55 bands 485 nm and 555 nm. Comparing the spectra in assigned to inplane CH deformation of benzimidazole
56 Figure S1A (a) to B (a), the disappearance of monomer and phenyl ring [17]. A band at 1030 cm1 may indicate
57 peak (410 nm) and the appearance of a broad peak at the presence of positive charges in polymer chains and
58 485 nm prove the deposition of P2AB polymer on ITO

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17 Fig. 2. (A) FTIR and (B) RAMAN spectra of P2AB and P2AB/AuNPs films on PGE.
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38 Fig. 3. FE-SEM imaging of (a) P2AB (b,c) P2AB/AuNPs at different magnifications (d) SEM-(BSE and SE) images of P2AB/AuNPs/
39 PGE surface, SEM-EDX mappings of e) gold and f) nitrogen on P2AB/AuNPs/PGE surface.
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42 also assigned to the CN stretching vibrations of the stretching, NH in-plane bending and C=N vibration of
43 aminophenyl group [16, 18]. the aromatic ring, respectively [20–21].
44 The Raman Scattering measurements were carried out
45 to characterize the P2AB, and P2AB/AuNPs coated PGEs
3.1.3 SEM-EDX
46 (Figure 2B). When compared the spectra of two polymers,
47 the prominent peaks appear in the presence of AuNPs, as The morphological structures of the P2AB and P2AB/
48 expected. In the spectrum of P2AB/AuNPs film, the peak AuNPs coated PGEs were examined by FE-SEM. As
49 at 958 cm1 is corresponding to CIO4 as counter ion [19]. shown in Figure 3 (a), P2AB film exhibits rough surface
50 The bands at 673 cm1 assigned to CC-C out of plane with a non-uniform distributed defect. On the other hand,
51 bending, the bands at 787 cm1 and 858 cm1 are corre- the FE-SEM images of P2AB/AuNPs (Figure 3 (b)) show
52 sponding to the CH out of plane bendings- and the peaks a layered coating with spherical particles, which has
53 at 1123 cm1 and 1186 cm1 are related to the CH in sizeable electroactive surface area. The spherical particles
54 plane bending [20]. The CC aromatic ring chain may be evidence of Au deposits on the surface of the
55 vibrations observed at 1020 cm1 and 1455 cm1 [20]. Also, electrode with the polymer. In order to understand
56 the bands occurring at 1266 cm1, 1345 cm1, 1534 cm1 whether or not Au exists, analysis of the surface was
57 and 1620 cm1 are attributed to CC stretching, the CN carried out by SEM with Backscattered and Secondary
58 Electron (BSE and SE) detectors (Figure 3 (d)) and EDX

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1 Au mapping (Figure 3 (e)). The contrast of Au on the P2AB/AuNPs is ten times higher than that of P2AB
2 surface can easily be seen in Figure 3 (d). From Figure 3 coated film. The higher pore resistance may be associated
3 (e), Au element appears intensively and almost homoge- with gold penetration into the pore of compact film.
4 neously, verifying the existence in the coating. Figure 3 (b)
5 shows the cluster size of Au nanoparticles, which are in
3.1.5 Dry Conductivity
6 the range of 30–400 nm. At higher magnification (Fig-
7 ure 3(c)), it is seen that the Au nanoparticles, in the range The four-probe technique was used to measure the dry
8 of 8 to 10 nm, agglomerate as the large gold particles. In conductivity values of the P2AB and P2AB/AuNPs films
9 addition, the EDX N mapping of P2AB/AuNPs coatings at room temperature.The P2AB conductivity value
10 (Figure 3(f)) indicates that the polymer deposited on the couldn’t be measured due to the out of range of the four-
11 electrode surface along with Au nanoparticles. probe conductivity device. Alternatively, the conductivity
12 value of P2AB/AuNPs film was determined as 1.3 3 104 S/
13 cm. The measurable change in conductivity could be the
3.1.4 EIS
14 result of both the presence of Au and the formation of
15 Electrochemical impedance spectroscopy (EIS) provides longer chains in the polymer.
16 detailed information on the impedance changes of an
17 electrode surface, using an equivalent circuit. Figure 4 and
18 inset show the Nyquist and Bode plots for P2AB 3.2 Electrocatalytic Reduction of H2O2 at the
19 homopolymer (Figure 4 (b)) and P2AB/AuNPs composite P2AB/AuNPs/PGE
20 (Figure 4 (c)) coated PGEs in 100 mM PBS (pH 6.5). To
3.2.1 Electrochemical Behavior of the P2AB/AuNPs/PGE
21 prove the modification of conducting films on PGE, the
in the Presence of H2O2
22 impedance spectrum of the bare PGE was also recorded
23 (Figure 4 (a)). EIS measurements were fitted using Electrochemical behaviors of P2AB and P2AB/AuNPs
24 ZSimpWin (Version 3.50). The inset in Figure 4 shows the coated electrodes were examined in 100 mM PBS of
25 equivalent circuits used to fit the experimental curves. pH 6.5 containing 1.0 mM H2O2 by cyclic voltammetry,
26 The fitting values for the equivalent circuit elements and scanning between 1,0 V and 1.0 V vs. SCE at a scan rate
27 c2 values are presented in Table S1. The terms Rs, Rpore of 100 mV s1. While the bare electrode does not respond
28 and Rct refer to the solution resistance, the pore resistance to H2O2 (Figure 5A (c)), there is a considerable increase
29 at the electrolyte polymer interface and the charge trans- in the reduction peak currents for the coated electrodes.
30 fer resistance at the polymer electrode interface, respec- When the redox peaks of coated electrodes are compared
31 tively. Q expressed the constant phase element (CPE), with each other, they exhibit almost similar behaviors as
32 which is used in place of double layer capacitance to seen in Figure 5A (a) and (b), but there is an increase in
33 describe the inhomogeneities [22]. According to Table S1, currents of both oxidation and reduction for P2AB/
34 P2AB coated PGE exhibits ten times lower Rct (and Rtotal) AuNPs coated electrode (Figure 5A (b)). It is indicated
35 value than that of bare PGE. This is attributed to the fact that P2AB polymer in composite film is oxidized with the
36 that the surface of the PGE was successfully modified by addition of H2O2 in PBS solution while H2O2 is reduced.
37 conducting polymer. However, Rpore (and Rtotal) value of Notably, the reduction current dramatically increases
38 about 0.8 V in the presence of AuNPs. This demon-
39 strates that AuNPs could efficiently catalyze the reduction
40 of H2O2 while increasing the conductivity of the P2AB
41 film. According to the literature [23], the mechanism for
42 H2O2 electroreduction on the P2AB/AuNPs coated elec-
43 trode could be expressed as following:
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45 H2 O2 þ e ! OHad þ OH ð1Þ
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47 OHad þ e ! OH ð2Þ
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2 OH þ 2 Hþ ! 2 H2 O ð3Þ
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Eventually, the modified electrode exhibits higher
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electrocatalytic activity toward H2O2 reduction due to the
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combination of AuNPs with P2AB.
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54 The effect of scan rates on the electrochemical
55 Fig. 4. Nyquist plots obtained in 100 mM PBS solution for coat- behavior of P2AB/AuNPs/PGE has been studied in
56 ings of (a) bare PGE and (b) P2AB (c) P2AB/AuNPs coated 100 mM PBS of pH 6.5 containing 2.0 mM H2O2. The
57 PGEs (Their Bode plots and equivalent circuits are also shown as peak currents were measured at varying scan rates from
58 inset). 10–300 mV s1. As seen in Figure 5B, the cathodic peak

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16 Fig. 5. (A) Cyclic voltammograms of (a) P2AB/PGE (b) P2AB/AuNPs/PGE (c) bare PGE in PBS containing 1.0 mM H2O2, v =
17 100 mV s1. (B) Cyclic voltammograms of the P2AB/AuNPs electrode in 100 mM PBS containing 2.0 mM H2O2 (pH 6.5) as a function
18 of scan rate (10–300 mV s1). (inset : Plot of peak currents vs. the square root of scan rate).
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30 Fig. 6. (A) Linear dependence of the response the current vs. the H2O2 concentration as a function of the pH of PBS (4.5, 5.5, 6.5, 7.5)
31 for P2AB/AuNPs/PGE (B) Amperometric i-t curves obtained from different concentration of H2O2 in 100 mM PBS of pH 6.5 at 0.8 V
32 for (a) P2AB/PGE (b) P2AB/AuNPs/PGE (inset is enlarged Figure 6B (a)) (C) Calibration for linearity range of H2O2 concentration
33 over P2AB/AuNPs/PGE with error bars (Error bars indicate the standard error of the mean (n = 3)).
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36 current increases with increasing scan rate for P2AB/ supporting electrolyte medium was determined as
37 AuNPs/PGE. Also, it is linearly proportional to the square 100 mM PBS of pH 6.5.
38 root of scan rate over a range from 10 to 300 mV s1 The current response of P2AB/AuNPs/PGE in the
39 (Figure 5B), indicating that the electrochemical response linear range was investigated by successive addition of
40 of the P2AB/AuNPs/PGE electrode is a typical diffusion- H2O2 with increasing concentrations into 100 mM PBS of
41 controlled process. pH 6.5 at the applied potential of 0.8 V (Figure 6B (b)).
42 When the response of P2AB homopolymer coated PGE
43 compared to that of P2AB/AuNPs/PGE (Figure 6B (a)), a
3.2.2 Amperometric Response of the P2AB/AuNPs/PGE
44 dramatic current increase was observed for P2AB/AuNPs/
to H2O2
45 PGE. Additionally, P2AB/AuNPs/PGE had a short re-
46 Considering the response of bare electrode to H2O2 in the sponse time about 6 seconds and an equal current step.
47 PBS solution (Figure 5 (c)), amperometric responses were Such fast response was attributed to a rapid electron
48 studied at the potentials between 0.8 V and 0.4 V using transfer between H2O2 and P2AB/AuNPs/PGE. Equal
49 P2AB/AuNPs/PGE by amperometric i-t curves. The better current steps indicate the stable catalytic properties of the
50 responding potential was determined as 0.8 V, and it was modified electrode to H2O2. The steady state current
51 preferred as an applied potential for determination of values were plotted against the bulk concentration of
52 H2O2. The pH effect of the P2AB/AuNPs/PGE was also H2O2 with error bars using P2AB/AuNPs/PGE (Fig-
53 investigated for catalytic activity to the H2O2 reduction. ure 6C). This electrode has a detection limit of 3.67 3 105
54 The current values were plotted against the bulk concen- M estimated at a signal to noise ratio of 3. The current
55 tration of H2O2 with error bars (Figure 6A). Notably, the response was linear with respect to H2O2 concentration
56 slope of calibration curve increases up to pH 6.5 and then over the range from 0.06 to 100 mM (R2 = 0.992) with a
57 decreases again (at pH 7.5). Therefore, the optimum sensitivity of 0.0395 A/M. The validation parameters
58 obtained for P2AB/AuNPs/PGE were summarized in

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1 Table 1 (A). The performance of P2AB/AuNPs/PGE was 50.0 mM concentration of glucose, ascorbic acid, dopa-
2 summarized comparatively with other H2O2 sensors based mine and uric acid cause negligible interference on the
3 on AuNP modified electrodes in the literature (Table S2). detection of H2O2 in 100 mM PBS of pH 6.5.
4 Accordingly, although the LOD of the fabricated elec- To confirm the validation of P2AB/AuNPs/PGE in the
5 trode could not be significantly improved, a large linear detection of H2O2 in real samples, the hair dye oxidant
6 range (0.06 mM–100 mM) was obtained that responded to sample (3 %) containing the specific concentrations of
7 the high H2O2 concentration. The long-term stability of H2O2 was added to PBS and analyzed under optimum
8 the P2AB/AuNPs electrode was investigated by storing it conditions obtained above by using the standart addition
9 at room temperature in the atmosphere. The current technique. As seen in Table 1B, the prepared electrode
10 response to 1.00 mM H2O2 was the same for three days shows good recovery for determination of different
11 and maintained more 53 % of its initial value after ten concentrations of H2O2.
12 days.
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4 Conclusions
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Table 1. (A) Regression data of the calibration curve for assay of
15 Poly(2-aminophenylbenzimidazole) (P2AB) film with
H2O2 by P2AB/AuNPs/PGE (B) Determination of H2O2 in real
16 sample (n = 3). gold nanoparticles (AuNPs) coated pencil graphite elec-
17 trode (PGE) was deposited by cyclic voltammetry in
18 (A) Calibration parameters of acetonitrile solution by a one-step route. Briefly, HAuCI4
19 P2AB/AuNPs/PGE was reduced to AuNPs in the presence of partially
20 Slope of calibration curve, (A/M) 0.0395 protonated 2-aminophenylbenzimidazole monomer in
21 Intercept (A) 1.18 3 105 acetonitrile. AuNPs facilitated the electron transfer proc-
22 SD (standard deviation) of calibration (A) 1.20 3 106 ess, which means that more polymer deposited on the
23 SD of slope, (A/M) 1.68 3 103 PGE. After optimization and characterization studies, the
24 SD of intercept, A 4.836 3 107 enzyme-free analysis of H2O2 was examined by using
Limit of detection (LOD) (M) 3.67 3 105
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Limit of quantification (LOQ) (M) 1.22 3 104
P2AB/AuNPs/PGE. It is determined that the sensor
26 Regression coefficient, R2 0.992 efficiently catalyzes the reduction of H2O2. The sizeable
27 electroactive surface area of P2AB/AuNPs/PGE improves
28 (B) Sample Added/mM Recovery/% RSD/% (n = 3) the analytical performance of sensor. Consequently,
29 a 1.5 110 3.5 P2AB/AuNPs/PGE can be used to as an alternative for
30 b 3.0 105 9.1 non-enzymatic H2O2 determination with a low detection
31 limit (3.67 3 105 M), wide linear range (0.060 to 100 mM)
32 and easy preparation. In future studies, this electrode can
33 For the selectivity test of P2AB/AuNPs/PGE, the be applied to various biological and environmental studies
34 interfering electroactive substances were, ascorbic acid, for analytical determination.
35 uric acid, glucose, and dopamine, selected in a similar
36 manner to those reported in the literature [7c, 24]. The
Acknowledgements
37 effect of substances on the response of modified electrode
38 is presented in Figure 7. At an applied potential of 0.8 V, The authors would like to thank the financial support
39 from the Hacettepe University Scientific Research Unit
40 (014 D12 601 001-805).
41
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