DEL RECURSO DE REVISIÓN

6.7.1 Sensors based on chemically and electrochemically synthesized Ni-based LDH for the detection of glucose

6.7.1.1 Equipments

Metal analyses of the LDHs obtained by chemical synthesis were performed by Varian 700-ES series Inductively Coupled Plasma–Optical Emission Spectrometer (ICP–OES) using solutions prepared by dissolving the samples in concentrated HNO3 and then properly diluted.

Coupled thermogravimetric (TGA) and differential thermal (DTA) analyses were performed with a Netzsch STA 449C apparatus, in air flow and heating rate of 10 °C min-1.

The XRD patterns of the chemical and electrosynthesized LDH films were recorded, after metallization with gold, using a Philips X’Pert Pro diffractometer with a Cu Kα radiation ( = 0.15415 nm) operating at 40 kV and 40 mA equipped with an X’Celerator detector. A step size 0.0170 2 degree with 20 s step scan was used for chemically prepared LDHs. The samples were analyzed directly introducing the Pt electrode plates (area =1 cm2) in the optical center of the diffractometer.

Atomic force microscopy (Solver-Pro, NT-MDT) was also used to provide information on particle size and shape. The samples of the chemically and electrochemically

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synthesized LDHs were deposited on Pt plates (area = 1 cm2). Atomic force microscope topographical images were collected using an NT-MDT solver scanning probe microscope in tapping mode.

6.7.1.2 Pt surface modification

The LDHs obtained by chemical route were synthesized by Researchers from National Research Council (CNR-ISOF) of Bologna and from the University of Perugia, according to the following method (all chemicals were of reagent grade purity, purchased by Sigma-Aldrich and used as received). Colloidal aqueous dispersions of Mg/Al, Mg/Fe, Ni/Al and Ni/Fe LDHs were prepared by the double-microemulsion method [8]. Briefly, two microemulsions designated A and B were prepared by dispersing 6.25 g of Cetyltrimethylammonium bromide, 18.1 mL of n-butanol, 7.75 mL of isooctane, and 6.75 mL of aqueous phase. The aqueous phase A consisted of 0.4 M M(NO3)2 (where M can be Mg or Ni) and 0.125 M M(NO3)3 (where M can be Al or Fe). The aqueous phase B was a NH3 solution (5.0 M for the Mg/Al and Mg/Fe LDH samples and 1.25 M for the Ni/Al and Ni/Fe LDH samples).

Equal volumes of the two microemulsions were mixed to obtain the precipitation of LDH nanoparticles (NPs) in the reverse micelles. The resulting system was aged at 70 °C for 16 h. Afterward, the particles were recovered by centrifugation (12,000 rpm, 10 min). The precipitate was washed with an ethanol–chloroform mixture (1:1 v/v) (3 x 30 mL) and then with water (3 x 30 mL). Finally, the precipitate was reconstituted in water (40 mL) to give a colloidal dispersion which was characterized in term of NPs amount by a gravimetric method ([NPs] = 3.5 mg/mL).

After 15 minutes of sonication, 1 µL of a NPs colloidal aqueous dispersion, thus corresponding to a deposited mass of 3.5 g, was deposited by drop-casting on pretreated Pt electrodes. Finally, the electrode was stored at r.t. for 24 h up to complete dryness. The electrochemically synthesized LDHs were obtained as described in Par. 6.3.1.1.

6.7.1.3 CV characterizations and amperometric detection of glucose

After the preparation of the electrodes described in the previous paragraph, both the chemically and electrochemically prepared LDHs were stabilized in 0.1 M NaOH, performing 20 CV cycles between +0.3 and +0.7 V in the case of the electrochemical

105 Ni/Al and the chemical Ni/Fe LDH, between +0.3 and +0.6 V for the electrochemical Ni/Fe LDH and between +0.3 and +0.75 V for the chemical Ni/Al LDH. The stabilization is carried out in order to activate the Ni redox system; between the very first cycles and the last ones, when the cycles become superimposable, the current slowly increases.

Mg/Fe and Mg/Al LDHs were characterized also by CV, performing 20 cycles in the same electrolytic solution between -0.8 and +0.75 V.

The analytical performances of the chemically and electrochemically synthesized Ni/Al and Ni/Fe LDHs were studied in 0.1 M NaOH; the electrode responses were evaluated recording CV curves at different scan rates (0.05 and 0.01 V s-1) for increasing glucose additions (0.1 mmol L-1) to define the working potential. After each addition the solution was stirred for a few minutes and then 3 CV cycles were recorded.

For the best materials in terms of sensitivity, also chronoamperometric curves were recorded for subsequent glucose additions, keeping the solution under stirring. From these curves the calibration graphs (current vs concentration) were obtained. The chosen potential for the measurements were +0.58 V for the electrochemical Ni/Fe and +0.65 V for the electrochemical Ni/Al LDH.

Nyquist plots were obtained recording impedance spectra for each coated electrode, at the peak potential (0.56 and 0.62 V for the electrochemically synthesized Ni/Fe and Ni/Al LDHs, respectively, and 0.61 and 0.66 V for the chemically synthesized Ni/Fe and Ni/Al LDHs).

The catalytic constants were calculated in the following way, according to the literature [9]. Equation 6.4 allows the determination of the rate constant of the reaction between an oxidizable substrate and the oxidized Ni centres in the LDH material:

(6.4)

where Icat is the catalytic current recorded at the Ni coated electrode in the presence of the substrate, IL is the limiting current recorded in the same conditions in the absence of an oxidizable substrate, and γ is defined as:

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where k is the reaction kinetic constant, t is time and c0 is the substrate bulk concentration. When γ exceeds 1.5, the second factor of Eq. (6.4) is almost equal to 0.5 and the equation can be reduced to:

_(6.6)

Accordingly, the slope of the plot Icat/IL as a function of c00.5t0.5 is equal to (k·0.5. For each electrode the Icat/IL versus c00.5t0.5 plot is linear when the value of c00.5t0.5 is between 5∙10−3 and 2∙10−2M0.5· s0.5, and this indicates a good adherence to the model. The chronoamperometric curves for those calculations were performed by recording the response of the modified electrode in absence of glucose and after several additions of analyte (each one corresponding to 0.05 mM) in the linear range. A pretreatment potential was firstly applied for 30 s, corresponding to a potential at which the Ni centres starts to be oxidized; the pretreatment was then followed by the application of the potential at which the electrocatalysis occurs, for 10 s. The potentials chosen were Epre = 0.61 V, E = 0.65 V for the electrochemical Ni/Al, Epre = 0.56 V, E = 0.58 V for the electrochemical Ni/Fe, Epre = 0.55 V, E = 0.63 V for the chemical Ni/Al LDH. The catalytic constant was not calculated for the chemical Ni/Fe LDH since the electrocatalysis occurs at potentials very close to the solvent discharge.

6.7.2 Co/Al LDH coated Pt for in flow amperometric detection of sugars

The apparatus for the flow injection analysis (FIA) consisted of a HPLC pump (LabFlow 1000), a six-way injection valve equipped with a 50 L loop, an electrochemical detector (Metrohm, VA 641 Model) interfaced with a personal computer. The electrochemical detector was constituted of 1 L wall-jet cell (Metrohm, 656 Model) in the three- electrode configuration (Ag/AgCl/ KCl 3.0 M as the reference, 2 mm diameter Au as the counter, and the working electrode) and a potentiostat for the amperometric measurements. In the HPLC configuration, the METROSEPCARB1 column (Metrohm) was inserted between the injection system and the detector. The eluent was a 0.1 M NaOH solution prepared starting from a 50 % (w/w) solution, previously-decarbonated, and, DD water. The flow rate was set at 1 mL min-1.

107 6.7.2.1 Pt surface modification

The electrodeposition was carried out on pretreated Pt electrodes, using the chosen condition for the potentiostatic synthesis described in section 6.3.1.1, for different times, namely 10, 20, 30 and 60 s. The modified electrodes were washed with DD water before performing the characterizations in CV. The electrocatalytic tests were performed for the LDHs electrosynthesized for 20 and 60 s.