Samples were then fixed on stubs and coated with a thin

Samples were then fixed on stubs and coated with a thin Vemurafenib manufacturer gold layer using a cool sputter-coater (Blazers SCD 005, Liechtenstein). The chitosan crosslinked with the chelating agent 8-hydroxyquinoline-5-sulphonic acid and glutaraldehyde and the microspheres were

obtained according to a procedure described in the literature (Vitali et al., 2008). The sensor was constructed as follows: 30 mg (15% w/w) of chitosan microspheres and 130 mg (65% w/w) of graphite powder were mixed in a small mortar for 20 min to form a homogeneous mixture and 40 mg (20% w/w) of Nujol was then added followed by mixing for another 20 min. The resulting modified carbon paste was tightly packed into a syringe and a copper wire was introduced into the other end for electrical contact. A bare carbon paste electrode (CPE), used for comparison purposes, was prepared as previously described, using GDC-0199 manufacturer only graphite powder and Nujol in the proportion of 65:35% w/w (Oliveira, Fernandes, & Vieira, 2006).

In a typical procedure for electrochemical measurements, 10.0 mL of the acetate buffer solution (pH 6.0) was transferred to a clean dry cell and successive additions of standard or sample solutions of Cu(II) were added by micropipette. The voltammetric procedure consisted of pre-concentration (accumulation), stripping (detection), and electrode regeneration steps. During the pre-concentration step, the electrode was immersed in the cell containing the supporting electrolyte and the metallic ion

solution. A negative controlled potential (−0.1 to −0.7 V) was applied to the sensor for a specified time (0–300 s). The solution was stirred using a magnetic stirring bar. Stripping voltammetry was then Exoribonuclease performed in the same cell with a sweeping square wave potential toward the positive direction (from −0.3 to 0.1 V), at frequencies of (f) 1.0–50 Hz, pulse amplitudes (a) of 10–50 mV and scan increments (ΔEs) of 1.0–10 mV, after successive additions of the analyte. The electrode cleaning step was performed by applying a positive potential under stirring. No de-aeration of solutions was required in any step. The sample and blank solutions were prepared following previously described procedures (Onianwa, Adetola, Iwegbue, Ojo, & Tella, 1999). Briefly, three samples of instant coffee (A, B and C) were obtained from local supermarkets in Florianópolis (Santa Catarina, Brazil). For the sample preparation, 1.0 g of instant coffee was weighed in triplicate in porcelain crucibles and mineralised in a muffle furnace at 550 °C for 20 h. The mineralisation step is necessary in order to eliminate the organic compounds present in the coffee sample, which can act as complexing agents for many metals (including Cu(II)) and can thus affect the results if present in the sample. The residue was dissolved with 0.2 mL of 1.0 mol L−1 nitric acid and diluted to 3.0 mL with acetate buffer solution (0.1 mol L−1, pH 6.0).

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