Electrochemical behaviour towards ABTS oxidation

The electrochemical behaviour of ABTS was studied on bare and aryl-modified GC electrodes using cyclic voltammetry (CV) and the rotating disk electrode (RDE) method. First, the CV curves of bare GC electrodes were recorded between 0 and 0.8 V in order to confirm the results obtained in the literature [106, 110]. As can be seen from Figure 4, the CVs showed well-defined one-electron reversible waves which correspond to the ABTS^2/ABTS^ redox couple and in addition, the electrochemical response of ABTS on bare GC was independent of pH. The redox potential of the ABTS^2/ couple was found to be 0.46 V, which is in good agreement with the previous report [110]. Further-more, the experiments showed that the peak potential separation (ΔE_p) was determined as 0.08 V in the solutions of various pH (pH=4–8), while Palmore et al. [106] reported that in the solution of pH 4, the value of ΔE_p was 0.110 V.

While the electrochemical behaviour towards the ABTS^2/ABTS^ redox probe on bare GC is pH independent, only the representative RDE voltammetry curves for ABTS oxidation on a bare GC electrode at pH 5 are shown in Figure 5. Current plateaux are observed on unmodified GC in the range of potentials between 0.55 and 0.75 V. The theoretical diffusion-limited current was calculated by the Levich equation (2) according to Ref. [167]:

2

where I_d represents diffusion-limited current, n is the number of electrons involved (n = 1), F is the Faraday constant (96485 C mol⁻¹), A is the electrode area (0.2 cm²), C is the concentration of ABTS in the bulk (0.5×10⁻⁶ mol cm⁻³), D is the diffusion coefficient of ABTS (3.4×10⁻⁶ cm² s⁻¹ [115] or 4.4×10⁻⁶ cm² s⁻¹ [114]), ν is the kinematic viscosity of the solution (0.01 cm² s⁻¹ [171]) and ω is the electrode rotation rate. It is relevant to note that in the paper [I], the theoretical Levich plot was calculated using the value of D as reported by Di Fusco and co-workers (D = 3.4×10⁻⁶ cm² s⁻¹ [115]). However, it was of special interest to see whether the new D value of ABTS (4.4×10⁻⁶ cm² s⁻¹ [114]) determined later by Zeng et al. could fit with the experimental results obtained in this study. As can be seen from Figure 5 insets 1,2, the oxidation current is equal to the theoretical diffusion-limited current while using the value of D = 3.4×10⁻⁶ cm² s⁻¹ [115] (see Figure 5, inset 1). The half-wave potential (E_1/2) of ABTS oxidation on bare GC was independent of pH and the E_1/2 value was 0.46 V, exactly the same as determined from the CV data of bare GC. This is an expected result considering the nearly reversible CV response of ABTS.

0.0 0.2 0.4 0.6 0.8

Figure 4. Cyclic voltammograms of a bare GC electrode in Ar-saturated 50 mM acetate buffer (pH 4 and 5) and 50 mM phosphate buffer (pH 6, 7 and 8) containing 0.5 mM ABTS and 0.1 M KCl.

v = 100 mV s^1.

Figure 5. RDE voltammetry curves for ABTS oxidation on a bare GC electrode in Ar-saturated 50 mM acetate buffer (pH 5), containing 0.5 mM ABTS and 0.1 M KCl at various rotation rates.

v = 10 mV s^1. The insets show the Levich plot at 0.6 V. The solid line corresponds to the theoretical Levich plot, where the D was used as (1) 3.4×10⁻⁶ cm² s⁻¹ [115]

or (2) 4.4×10⁻⁶ cm² s⁻¹ [114]), respectively.

Figures 6a,c,e,g,i and Figures 6b,d,f,h,j show the CV curves and RDE voltammetry curves towards the ABTS^2/ redox probe on aryl-modified GC electrodes, respectively. The voltammetric response of bare GC is given for comparison. In addition, the electrochemical data for the ABTS^2/ couple of

bare GC and differently modified GC electrodes after modification by Procedures (a) and (c) were determined from the CV curves presented in Figures 6a,c,e,g,i and are given in Table 1. One of the parameters, the relative blocking efficiency (I_rel), was used for aryl-modified GC electrodes and was defined analogously to Ref. [163]:

rel = ^pa with a film

pa for a bare GC × 100% (3)

where Ipa is the intensity of the anodic peak current. It should be noted that while using the modification Procedure (b) for GC/NP and reduced GC/NP electrodes and Procedure (d) for GC/CP electrodes, the blocking effect was so strong that the CV peaks were not discernible (see Figures 6a,c,e,g,i) and for these electrodes the parameters to characterise their barrier properties towards the ABTS^2/ redox couple could not be determined.

According to the values given in Table 1, the ABTS^2/ redox system exhibits a significant increase of the ∆E_p value and decrease in peak current for NP-modified GC electrode (Procedure a) as compared to bare GC in all solutions. It should be noted that the increase in the separation of peak potentials demonstrates the inhibition of the charge transfer process. But in general, the electrochemical behaviour of ABTS on the GC/NP electrode (Procedure a) was almost the same in the solutions of different pH.

Interestingly, the reduced GC/NP (Procedure a) and GC/CP (Procedures c) electrodes showed a rather similar behaviour in the charge transfer process (at pH 4) because the values of ∆Ep were almost the same (see Table 1).

As can be seen from Table 1 and Figure 6, amongst other aryl-modified electrodes studied, some differences were observed in the electrochemical behaviour of ABTS on reduced GC/NP electrodes in different pH solutions as well (Figures 6a,c,e,g,i). It is known that the pKa of aniline is 4.6 [42, 172], therefore at pH 4 and 5 the AP groups on themodified electrode are protonated.

At higher pH (pH >> pKa) the AP groups are deprotonated and the electron transfer of the ABTS^2/ couple is somewhat blocked. Furthermore, the reduced GC/NP electrode (Procedure a) showed the lowest barrier properties for the ABTS oxidation in the solutions of different pH (pH=4–8), especially as compared with GC/NP electrodes (Procedure a). This might indicate that the aryl layer of NP is more compact than the reduced NP film where the aminophenyl groups are present (as shown by XPS in Section 6.1.2).

0.0 0.2 0.4 0.6 0.8 Reduced GC/NP (Procedure a) GC/NP (Procedure b) Reduced GC/NP (Procedure b) GC/CP (Procedure c) Reduced GC/NP (Procedure a) GC/NP (Procedure b) Reduced GC/NP (Procedure b) GC/CP (Procedure c) Reduced GC/NP (Procedure a) GC/NP (Procedure b) Reduced GC/NP (Procedure b) GC/CP (Procedure c) Reduced GC/NP (Procedure a) GC/NP (Procedure b) Reduced GC/NP (Procedure b) GC/CP (Procedure c) Reduced GC/NP (Procedure a) GC/NP (Procedure b) Reduced GC/NP (Procedure b) GC/CP (Procedure c) Reduced GC/NP (Procedure a) GC/NP (Procedure b) Reduced GC/NP (Procedure b) GC/CP (Procedure c) Reduced GC/NP (Procedure a) GC/NP (Procedure b) Reduced GC/NP (Procedure b) GC/CP (Procedure c) Reduced GC/NP (Procedure a) GC/NP (Procedure b) Reduced GC/NP (Procedure b) GC/CP (Procedure c) Reduced GC/NP (Procedure a) GC/NP (Procedure b) Reduced GC/NP (Procedure b) GC/CP (Procedure c) Reduced GC/NP (Procedure a) GC/NP (Procedure b) Reduced GC/NP (Procedure b) GC/CP (Procedure c) GC/CP (Procedure d)

I / mA

E vs SCE / V

Figure 6. (a,c,e,g,i) CVs and (b,d,f,h,j) RDE voltammetry curves for bare and aryl-modified GC electrodes in Ar-saturated buffer solutions containing 0.5 mM ABTS and 0.1 M KCl at different pH (4–8). (a,c,e,g,i) v = 100 mV s^1 and (b,d,f,h,j) v = 10 mV s^1 and  = 1900 rpm.

The electrochemical results are rather controversial compared with literature.

Namely, Lyskawa and Bélanger [173] reported the electrochemical behaviour towards ferricyanide reduction of Au electrodes modified with NP or aminophenyl groups by in situ generated diazonium salt reduction. The experimental results revealed that the electrochemical response of the Fe(CN)₆^3/4 redox system was more suppressed for AP-modified Au electrodes than for NP-modified Au electrodes [173] and therefore it was further concluded that the aminophenyl layer was more compact than the NP layer [40].

Table 1. Comparison of the CV results of bare GC and aryl-modified GC electrodes, using the modification Procedure a (one cycle) and c (two cycles). CVs were recorded in Ar-saturated 0.1 M KCl solution of various pH (pH=4–8) containing 0.5 mM ABTS.

v = 100 mV s^1.

1 These electrodes were modified by Procedure (a); ² This electrode was modified by Procedure (c); n.d. – not detectable

According to the CV and RDE results, the largest pH effect was observed for the GC/CP electrodes (both Procedures c and d). In case of GC/CP electrode modified by Procedure (c), the oxidation peak of ABTS was clearly visible at pH 4 (Figure 6a), while at pH 5 (Figure 6c) the electrochemical response of ABTS was more suppressed and even more at pHs 6–8 (Figures 6e,g,i).

Therefore, the electrochemical parameters for GC/CP electrode (Procedure c) at pH>5 could not be determined as can be seen in Table 1. A similar behaviour of CP-modified electrodes has been observed by Saby et al. [39] and Abiman et al.

[174] for the Fe(CN)63/4 redox system in buffer solutions of differing pH. The authors revealed that as the pH of the solution increased, the peak current decreased significantly at the CP-modified electrode. This kind of blocking behaviour was explained by the surface pKa value of CP groups. Saby et al. [39]

reported that the apparent pKa of the 4-carboxyphenyl grafted layer shifted to a lower value (pKa = 2.8) than the pKa of solution based benzoic acid (pKa = 4.2).

It can be said that the CP groups on GC surface are deprotonated at the pHs studied according to the estimated pK_a of 2.8 [39] or 3.25 [174] and thus repel the negatively charged ABTS.

In order to get a better overview of the blocking effect, the differences of the currents of ABTS oxidation at 0.6 V between the electrodes of various aryl films were taken for comparison with that of bare GC. The respective current values are given in Table 2. As can be seen from Table 2, the effect of pH on ABTS oxidation was rather negligible in case of GC/NP (Procedures a,b) and reduced GC/NP (Procedure a) electrodes. Also the electrochemical behaviour towards the ABTS^2/ redox probe on the GC/NP electrode (Procedure a) was almost the same in the solutions of different pH (see Figure 6). However, the oxidation current values for GC/CP electrode (Procedure d) were higher at pH 4 than in other solutions (pH=5–8), where the current values were almost the same (this was discussed above). The same tendency was also noticed for the reduced NP film by applying Procedure (b).

Table 2. Comparison of oxidation current (I, mA) values at the potential of 0.6 V for a bare and aryl-modified GC electrodes obtained by the RDE method in Ar-saturated solutions of various pH (pH=4–8) containing 0.5 mM ABTS and 0.1 M KCl.  = 1900 rpm, v = 10 mV s^1.

Electrode ABTS oxidation current (mA)

pH 4 pH 5 pH 6 pH 7 pH 8 Bare GC 0.0399 0.0397 0.0399 0.0386 0.0390 GC/NP (Procedure a) 0.0109 0.0096 0.0120 0.0118 0.0077 GC/NP (Procedure b) 0.0002 0.0004 0.0002 0.0002 0.0002 Reduced GC/NP (Procedure a) 0.0270 0.0214 0.0153 0.0155 0.0113 Reduced GC/NP (Procedure b) 0.0018 0.0009 0.0001 0.0001 0.0011 GC/CP (Procedure c) 0.0283 0.0158 0.0130 0.0082 0.0080 GC/CP (Procedure d) 0.0134 0.0048 0.0048 0.0044 0.0069 The blocking action towards ABTS response increased with the number of modification cycles. The largest blocking effect towards ABTS was observed for a GC/NP electrode (Procedure b) in the solution of different pH (shown in Figure 6), but also for reduced GC/NP (Procedure b) electrodes as can be seen from Table 2. The GC electrodes modified with NP, CP and AP groups using Procedures (b) and (d) showed a similar response towards the ABTS^2/ couple at various pHs and yielded a stronger blocking effect as compared to the electrodes modified by Procedures (a) and (c). This is obviously related to the differences in the surface concentration of aryl groups.

In addition, the barrier properties of the aryl-modified electrodes can be explained by electrostatic interaction between charged groups and ionic redox probes according to the study by Schauff et al. [122] and as discussed in

Section 4.3. Besides the electrostatic interactions the barrier properties are strongly influenced by surface coverage of the modifier film. The structure of the aryl films has not been studied in the present work. However, previous investigations have shown that the blocking properties of aryl films depend on the modification conditions and even on the post-deposition treatment [8]. In addition, the presence and distribution of pinholes in the film might considerably influence its barrier properties. All these aspects are important for the practical application of aryl-modified electrodes in electroanalysis.

6.2. Surface and electrochemical properties of GC and