RESUMO
A selective and simple analytical method for the trace level determination of carbofuran in complex environmental and biological samples was developed based on immunoaffinity extraction (IAE) followed by on-line preconcentration and HPLC/UV analysis of the purified extract. The immunosorbent for IAE was prepared by sol-gel encapsulation of monoclonal anti-carbofuran antibodies, and was fully characterized for capacity, repeatability, binding strength, binding kinetics and cross-reactivity. Method performance was evaluated with two different types of difficult samples: dam water and methanolic extracts of epithelial cervical-uterine tissue. Linear behavior and quantitative recoveries were obtained from the analysis of samples spiked with carbofuran at 0.2-4 ng/mL (dam water, 50 mL samples) and 10-40 ng/mL (biological tissue extract, 2 mL samples). RSD (n=7) and detection limits were, respectively, 10.1% (spike 0.40 ng/mL) and 0.13 ng/mL for dam water; 8.5% (spike 20 ng/mL) and 5 ng/mL for the biological tissue extract. The excellent sample purification achieved with the IAE column allows precise and accurate determination of carbofuran in complex matrices, even when using non-selective UV detection in the chromatographic analysis.
Assuntos
Carbofurano/análise , Colo do Útero/química , Fracionamento Químico/métodos , Cromatografia de Afinidade/métodos , Poluentes Químicos da Água/análise , Adsorção , Anticorpos Monoclonais/química , Calibragem , Cromatografia Líquida de Alta Pressão , Epitélio/química , Feminino , Géis , Humanos , Proteínas Imobilizadas/química , Inseticidas/análise , Limite de Detecção , Metanol/química , Transição de Fase , ÁguaRESUMO
Living organisms are subject to stress, and among these stressors, heavy metals exposure triggers accumulation of sulfur metabolites. Among these metabolites, glutathione and phytochelatins are found in several organisms, such as Euglena gracilis. Pre-exposing E. gracilis to low concentrations of Hg2+ generates a population with resistance to even 0.2 mM Cd2+, and this resistance relies partly on phytochelatins. p38 MAPK is stimulated by stress and is involved in apoptotic as well as survival mechanisms. In this study, we explored its participation in heavy metal-induced stress and its possible role in sulfur metabolite accumulation. We found that about 51% of the E. gracilis pretreated with Hg2+ becomes resistant to Cd2+ and proliferates despite the presence of this metal. The accumulation of the sulfur metabolites gamma-glu-cys, glutathione and phytochelatin 2 displayed cyclic patterns that were disturbed by a challenge with Cd2+. We observed a p38 MAPK-like activity that was stimulated by acute or chronic heavy metal exposure, and its inhibition by SB203580 slightly diminished the accumulation of sulfur compounds. p38 MAPK inhibition also affected basal levels of glutathione in either pretreated or control cells. Thus, it appears that p38 MAPK mediates redox stress component of the signal pathway induced by heavy metals.