Development of an Ethanol Biosensor Based on Silver Nanoparticles/Polyaniline/Graphite/Epoxy Composite for Friendly Analytical Application

Electrochemical biosensors for measuring ethanol were developed in this study. Silver nanoparticles were incorporated to increase sensibility. Firstly, the transducer element was prepared with silver nanoparticles/polyaniline/graphite/epoxy composite (AgNPs/PANI/GEC) where the enzymes alcohol oxidase (AOD) and horseradish peroxidase (HRP) were immobilized by adsorption. The composition of the immobilized solution was indicated by an experimental design (85% of the bi-enzymatic solution, 10% of albumin and 5% of glutaraldehyde). The immobilization method adopted in this study showed to be highly reproducible. The values of variance and standard deviation were low (0.003 and 0.053, respectively—means of three electrodes). The linear range was 0–30 g/L (R2 = 0.983) and the sensitivity 0.004 mA L/g. A second biosensor was made with a transducer prepared with AgNPs/PANI/GEC. A chitosan film was applied over the adsorbed enzymatic solution to avoid desorption of the immobilized enzymes. The AgNPs/PANI/GEC electrodes were characterized using cycle voltammetry and the composite surface by scanning electron microscopy. The calibration for ethanol samples with this second composite fitted in a range of 0.0–0.35 g/L (R2 = 0.984). Square rate voltammetry was the electrochemical analytical method used to obtain the amperometric calibration curves

Decolorization of a chromophore molecule with immobilized horseradish peroxidase / Descoloração de uma molécula de cromóforo com peroxidase de rábano imobilizada

The enzymes can modify some effluent characteristics in order to increase the degradability, or the bioconversion of liquid  effluents. The oxireductases, laccases and peroxidases have been used due their high potentiality in many environmental treatments of natural and synthetic organic compounds as dyes, phenols and polyphenolics molecules. The performance of immobilized horseradish peroxidase on aminopropyl glass beads was investigated in this work in a decolorization reaction of methylene blue colorant. The experiments were conducted in batch conditions during 3 hours, with different aqueous solutions of peroxide hydrogen (H2O2) concentration solutions (2-10 mg/L), methylene blue (ME)  (5-20 mg/L) and the pH in the range from 4 to 8, according an experimental design proposed by the software STATISTICA®. After 3 hours of treatment the reduction of  the color was 60% when comparing to the original color and 50% when the immobilized enzymes were reused in five sequential batch treatment cycles for a 10 mg/L of H2O2 solution, 20 mg/L of ME  and pH 8.0.  Working in continuous process with two microreactors in series the system showed a good performance with 97% of decolorization in the first 15 minutes. After 1 hour of continuous treatment the percentage of the color removing was around 70%.

Characterization of an hrp-aox-polyaniline-graphite composite biosensor

Nowadays there is an increasing demand to develop new and robust biosensors in order to detect low concentrations of different chemicals, in practical and small devices, giving fast and confident responses. The electrode material was a polyaniline-graphite-epoxy composite (PANI/GEC). Alcohol oxidase (AOX) and horseradish peroxidase (HRP) enzymes were immobilized and the responses were tested by cyclic voltammetry. The conductivities for the composites of graphite/polyaniline were determined. The cyclic voltammograms allowed detecting ethanol in pure diluted samples in a range from 0.036 to 2.62 M. Differential scanning calorimetry (DSC) and thermal gravimetry analysis (TGA) were used to verify the thermal characteristics of the composites (0, 10, 20, 30 and 100 % of graphite). The Imax value was determined for the dual enzyme biosensor (0.0724 mA), and the Kapp m as 1.41 M (with R2 =0.9912)