RESUMO
We report the synthesis and characterization of a new hybrid magnetic composite formed by the enveloping of magnetic iron oxide nanoparticles (γ-NP) with chains of the conductive polymer PEDOT, and its use for the efficient separation of DNA molecules from complex biological samples, allowing the high yield separation of a pure and high-quality DNA fraction. The successful formation of the γ-NP/PEDOT composite was confirmed by Fourier transform infrared spectroscopy, scanning electron microscopy, UV visible spectroscopy (UV-Vis), and magnetic hysteresis loop measurements. The nanocomposites showed an excellent capacity of DNA adsorption (Qe â¼ 248 mg/g) in a model system consisting of salmon sperm DNA. When the γ-NP/PEDOT was used in protocols to extract the DNA from complex samples, the corresponding yield was in the range of 6.4 µg (blood) and 7.3 µg (bacteria), as evaluated quality by UV-Vis, PCR analysis, and electrophoresis assays. We also established that the captured DNA does not need to be detached from the nanocomposite for use as seeding material in PCR amplification experiments. These results and the simplicity of the protocols indicate that the γ-NP/PEDOT composite is a promising DNA absorbent, being competitive with the commercially available magnetic purification kits.
Assuntos
Nanocompostos , Compostos Bicíclicos Heterocíclicos com Pontes , DNA/genética , Polímeros , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
We report the preparation of poly(3,4-ethylene dioxythiophene) (PEDOT)-modified polyvinylidene fluoride electrospun fibers and their use as a novel adsorbent material for the removal of the anionic dye Methyl Orange (MO) from aqueous media. This novel adsorbent material can be used to selectively remove MO on a wide pH range (3.0-10.0), with a maximum capacity of 143.8 mg/g at pH 3.0. When used in a recirculating filtration system, the maximum absorption capacity was reached in a shorter time (20 min) than that observed for batch mode experiments (360 min). Based on the analyses of the kinetics and adsorption isotherm data, one can conclude that the predominant mechanism of interaction between the membrane and the dissolved dye molecules is electrostatic. Besides, considering the estimated values for the Gibbs energy, and entropy and enthalpy changes, it was established that the adsorption process is spontaneous and occurs in an endothermic manner. The good mechanical and environmental stability of these membranes allowed their use in at least 20 consecutive adsorption/desorption cycles, without significant loss of their characteristics. We suggest that the physical-chemical characteristics of PEDOT make these hybrid mats a promising adsorbent material for use in water remediation protocols and effluent treatment systems.
Assuntos
Poluentes Químicos da Água , Adsorção , Compostos Azo , Compostos Bicíclicos Heterocíclicos com Pontes , Concentração de Íons de Hidrogênio , Cinética , Polímeros , Polivinil , TermodinâmicaRESUMO
When fluorophores attach to nanostructured films of intrinsically conducting polymers (ICPs), a quenching of their fluorescence may occur. We have exploited these characteristics for the development of polymeric films that can be used in a simple and efficient molecular diagnosis protocol based on the selective detection of nucleic acids. Our procedure rests on the fact that the fluorescence of 6-carboxyfluorescein-labeled single-stranded DNA (FAM-ssDNA) probes is quenched upon their immobilization on nanostructured ICP - polypyrrole (PPY) and polyaniline (PANI) - films deposited on polyethylene terephthalate (PET) substrates. Hybridization occurs whenever a sample with the complementary sequence is brought in contact with the immobilized probe, with the newly formed ds-DNA chains detaching from the flexible polymeric film and causing the restoration of the fluorescence. This sensing system exhibits a low background signal that depends on both the thickness and hydrophobicity of the films. As a model system, we used a FAM-ssDNA probe specific for the Leishmania infantum parasite. The results confirm this procedure as a simple, fast and highly sensitive scheme for the recognition of the target DNA, with a detection limit of the 1.1â¯nM and 1.3â¯nM for the PPY/PET and PANI/PET films, respectively. In addition, this biosensor has excellent stability and exhibits a good and reproducible performance even when used for the direct detection of ssDNA in relatively complex biological samples.