RESUMO
Phytochelatins (PCs) are peptides that play an important role in homeostasis and detoxification of heavy metal in plants. Furthermore, they have been proposed as earlier potential biomarkers of aquatic pollution by heavy metals. Nowadays, several researchers have reported on current methods for quantification of glutathione (GSH) and the PCs (phytochelatin 2, phytochelatin 3, phytochelatin 4) quantification in plants. However, no method has reported the uncertainty of the measurement, which helps to improve the accuracy and quality assurance in the PC quantification. In this work, a new methodology using ultra-high-performance liquid chromatography coupled to mass spectrometry (UPLC-MS) to measure with high precision and accuracy the PCs in aquatic plants, was validated. Selectivity, linearity, limit of detection, limit of quantification, precision, trueness and uncertainty estimation were examined as parts of the method validation. The described method shows excellent linearity in different ranges for all analytes with coefficients of determination higher than 0.99. The relative standard deviation for intra-day precision was <3% and for inter-day <10%. All LOD and LOQ analytes ranged from 0.02 to 0.08 µg ml-1, and from 0.03 to 0.09 µg ml-1, respectively. The recoveries varied from 61% to 89%. In order to obtain an interval of results with the highest confidence levels, the uncertainty associated with the measurements was evaluated. The calibration curve (>50%) and recovery (19-44%) were the most important contributors to the total uncertainty. The proposed method was applied to quantify GSH and PCs in the aquatic plants Lemna gibba L., Myriophyllum heterophyllum Michx., Arenaria paludicola and Hydrocotyle ranunculoides L. fil., showing statistical differences in the mass fraction of the analytes.
Assuntos
Fitoquelatinas , Espectrometria de Massas em Tandem , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , IncertezaRESUMO
The elimination of organic contaminants in highly complex wastewater was tested using a combination of the techniques: electrocoagulation with aluminum electrodes and phytoremediation with Myriophyllum aquaticum. Under optimal operating conditions at a pH of 8 and a current density of 45.45 A m(-2), the electrochemical method produces partial elimination of contaminants, which was improved using phytoremediation as a polishing technique. The combined treatment reduced chemical oxygen demand (COD) by 91%, color by 97% and turbidity by 98%. Initial and final values of contaminants in wastewaters were monitored using UV-vis spectrometry and cyclic voltammetry. Finally, the morphology and the elemental composition of the biomass were characterized with using scanning electron microscopy (SEM) and energy dispersion spectroscopy (EDS). The presence of Al in the roots of plants in the system indicates that the aluminum present in the test solution could be absorbed.