RESUMO
This work has the goal of developing and evaluating a reusable fluorescent nanosensor for detection of Cu(ii) ion in aqueous solution, based on the immobilization of rhodamine B in silica nanoparticles prepared according to a modified Stöber method. In order to do this, a standard ammonium hydroxide ethanolic solution was mixed to ethanol under constant stirring, followed by the addition of tetraethoxysilane (TEOS). To immobilize the fluorescent reagent in the silica nanoparticles, rhodamine B ethanolic solution was added to the reacting mixture at different times (2; 3; 4 and 5 h) after starting the synthesis (which always lasts 7 h). The nanosensor obtained with the addition of rhodamine B after 5 h of synthesis showed the best sensitivity, measured as the fluorescence quenching, which was proportional to Cu(ii) ion. The nanosensor was selective to Cu(ii) ions and showed a linear range from 2.0 to 12.0 µmol L-1, detection limit of 0.40 µmol L-1, quantification limit of 1.3 µmol L-1, response time of 50 s, being possible to be reused 3 times. The nanosensor was applied to the determination of Cu(ii) in sugar cane spirit and the results obtained did not show significant differences from those obtained by atomic absorption spectrometry at a confidence level of 95%.
RESUMO
The efficiency of three different depletion methods to remove the most abundant proteins, enriching those human serum proteins with low abundance is checked to make more efficient the search and discovery of biomarkers. These methods utilize magnetic nanoparticles (MNPs), chemical reagents (sequential application of dithiothreitol and acetonitrile, DTT/ACN), and commercial apparatus based on immunoaffinity (ProteoMiner, PM). The comparison between methods shows significant removal of abundant protein, remaining in the supernatant at concentrations of 4.6±0.2, 3.6±0.1, and 3.3±0.2µgµL-1 (n=3) for MNPs, DTT/ACN and PM respectively, from a total protein content of 54µgµL-1. Using GeLC-MS/MS analysis, MNPs depletion shows good efficiency in removing high molecular weight proteins (>80kDa). Due to the synergic effect between the reagents DTT and ACN, DTT/ACN-based depletion offers good performance in the depletion of thiol-rich proteins, such as albumin and transferrin (DTT action), as well as of high molecular weight proteins (ACN action). Furthermore, PM equalization confirms its efficiency in concentrating low-abundant proteins, decreasing the dynamic range of protein levels in human serum. Direct comparison between the treatments reveals 72 proteins identified when using MNP depletion (43 of them exclusively by this method), but only 20 proteins using DTT/ACN (seven exclusively by this method). Additionally, after PM treatment 30 proteins were identified, seven exclusively by this method. Thus, MNPs and DTT/ACN depletion can be simple, quick, cheap, and robust alternatives for immunochemistry-based protein depletion, providing a potential strategy in the search for disease biomarkers.
Assuntos
Proteínas Sanguíneas/isolamento & purificação , Fracionamento Químico/métodos , Nanopartículas de Magnetita/química , Acetonitrilas/química , Proteínas Sanguíneas/análise , Proteínas Sanguíneas/química , Cromatografia de Afinidade , Ditiotreitol/química , Humanos , Tamanho da Partícula , Fatores de TempoRESUMO
This study describes a method for non-destructive detection of adulterated glibenclamide tablets. This method uses near infrared spectroscopy (NIRS) and fluorescence spectroscopy along with chemometric tools such as Soft Independent Modeling of Class Analogy (SIMCA), Partial Least Squares-Discriminant Analysis (PLS-DA) and Unfolded Partial Least Squares with Discriminant Analysis (UPLS-DA). Both brand name (Daonil) and generic glibenclamide tablets were used for analysis. The levels of glibenclamide in each type of tablet were evaluated by derivative spectrophotometry in the ultraviolet region. The results obtained from the NIR and fluorescence spectroscopy along with those obtained from multivariate data classification show that this combined technique is an effective way to detect adulteration in drugs for the treatment of diabetes. In the future, this method may be extended to detect different types of counterfeit medications.