RESUMO
An automated multi-pumping flow system is proposed for the chemiluminometric determination of ascorbic acid in pharmaceutical formulations, relying on the ability of semiconductor nanocrystals to generate short-lived reactive species upon photo-irradiation. A photo-unit based on visible-light-emitting diodes is used to photo-excite cadmium telluride (CdTe) quantum dots capped with glutathione, leading to the generation of radicals that react with luminol under alkaline conditions, yielding the chemiluminescence. Ascorbic acid acts as a radical scavenger, preventing the oxidation of luminol, thus ensuring a concentration-dependent chemiluminescence quenching. After system optimization, a linear working range of 5.0 × 10(-7) to 5.0 × 10(-6) mol/L ascorbic acid (r = 0.9967, n = 5) was attained, with a detection limit of 3.05 × 10(-7) mol/L and a sampling rate of 200/h. The flow system was applied to the analysis of pharmaceutical formulations and the results were in good agreement with those obtained by the reference titrimetric procedure (RD < ± 4.3%, n = 7).
Assuntos
Ácido Ascórbico/análise , Compostos de Cádmio/química , Glutationa/química , Pontos Quânticos , Telúrio/química , Química Farmacêutica , Medições Luminescentes , Luminol/química , Estrutura Molecular , Processos Fotoquímicos , Espécies Reativas de OxigênioRESUMO
Actin cytoskeleton disruption in host cells has been demonstrated for PTPases from pathogenic microorganisms. In this work, we analysed whether the secreted acid phosphatase from Entamoeba histolytica has phosphotyrosine phosphatase activity and the possibility that this activity may participate in damaging host cells. The secreted acid phosphatase of E. histolytica, which catalyses p-nitrophenyl phosphate hydrolysis at acid pH values, was found to have phosphotyrosine phosphatase activity. The enzymatic properties of phosphotyrosine phosphatase and acid phosphatase were virtually identical and included: Km values of 10 x 10(-4) M, no requirement for divalent cations, and sensitivity to molybdate, vanadate, and tungstate. The phosphotyrosyl phosphatase activity caused significant levels of cell rounding and detachment correlating with disruption of the actin stress fibres in HeLa cells. Thus, our data suggest that secreted phosphotyrosine phosphatase could play a cytotoxic role during amoebic infection.