RESUMO
We describe a detailed study on the possibility of analyzing aminated pesticides and metabolites using pre-column derivatization with fluorescein isothiocyanate (FITC) and their subsequent separation and detection by cyclodextrin capillary electrophoresis-laser-induced fluorescence (CE-LIF) detection. Different variables affecting the derivatization reaction (pH, FITC concentration, reaction time and temperature) and those related with the separation itself (buffer concentration, addition of various organic modifiers, pH, applied voltage and injection time) were studied. The limit of detection obtained was between 0.45 and 3.48 microg litre(-1) showing a relative standard deviation between 0.26 and 2.08% at a concentration level of 50 microg litre(-1).
Assuntos
Aminas/análise , Eletroforese Capilar/métodos , Fluoresceína-5-Isotiocianato , Corantes Fluorescentes , Praguicidas/análise , Espectrometria de Fluorescência/métodos , Aminas/química , Ciclodextrinas , Estrutura Molecular , Praguicidas/química , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Poluentes Químicos da Água/análiseRESUMO
The aim of the present study was to develop a quantitative-competitive PCR (QC-PCR) method to detect DNA from transgenic herbicide-resistant (roundup ready, RR) soybean and maize. Since no QC-PCR system for the quantification of RR maize had been published at the time of writing, a specific competitor DNA for transgenic event was developed. For the QC-PCR of RR-soybean, a commercially available competitor was employed. These internal standards were calibrated by coamplifying with mixtures containing RR-soybean and maize DNAs. The calibrated QC-PCR systems were applied to certified RR-soybean and maize flour mixtures in order to demonstrate their suitability not only for the quantification of the glyphosate resistance traits in DNA matrices, but also in practically relevant samples. In addition, a special focus of the present work was to compare the detection of QC-PCR products by slab gel and CGE with UV detection. CGE permitted the precise detection of transgenic events also below the equivalence points; while in slab gel electrophoresis, due to the low sensitivity the quantification of genetically modified DNA was allowed only at the equivalence point.