RESUMO
Rising interest in ellagic acid (EA) present in functional foods is supported by its antimutagenic, anticarcinogenic, antiviral, antibacterial and antioxidative effects. The present approach presents for the first time the determination of ellagic acid and other phenolics in wines by miniaturized solid phase extraction prior to capillary zone electrophoresis (CZE) with UV. The extraction was performed using a home-made miniaturized pipette tip column. The procedure allowed a significant reduction in conditioning/sample/washing/elution volumes. The effects of important factors affecting the extraction efficiency as well as electrophoretic performance were investigated to acquire optimum conditions. The analytes were separated within 10 min with a BGE containing 30 mmol/L sodium tetraborate 10% v/v MeOH pH 9.10. The optimized method was applied to the determination of ellagic acid in commercial and pilot-scale wines. Indeed, the content of EA was correlated with viticultural parameters such as grape varietal, production area, and aging conditions (oak wood guard and glass bottle ward). In order to validate the results, a comparison between the CZE and HPLC data was made.
Assuntos
Eletroforese Capilar , Ácido Elágico/análise , Vinho/análise , Cromatografia Líquida de Alta Pressão , Eletroforese Capilar/métodos , Análise de Alimentos , Qualidade dos Alimentos , Vitis/química , MadeiraRESUMO
Mercury (Hg) is a major environmental pollutant that can be disposed to the environment by human activities, reaching crops like vineyards during irrigation with contaminated waters. A 2-year study was performed to monitor Hg variations during reproductive and vegetative stages of vines after Hg supplementation. Variations were focused on total Hg concentration, the molecular weight of Hg fractions and Hg-proteins associations in roots, stems and leaves. Total Hg concentrations increased during reproductive stages and decreased during vegetative stages. Variations in length of these stages were observed, according to an extension of the vegetative period. Six months post Hg administration, in roots, stems and leaves, initial Hg proteic fractions of 200 kDa were catabolized to 66 kDa fractions according to a transition from reproductive to vegetative stages. However, 24 months after Hg supplementation, the 66 kDa Hg proteic fraction was continuously determined in a prolonged senescence. Accordingly, the identified proteins associated to Hg show catabolic functions such as endopeptidases, hydrolases, glucosidases and nucleosidases. Stress associated proteins, like peroxidase and chitinase were also found associated to Hg. During the reproductive periods of vines, Hg was associated to membrane proteins, such as ATPases and lipid transfer proteins, especially in roots where Hg is absorbed.
Assuntos
Poluentes Ambientais/toxicidade , Mercúrio/toxicidade , Folhas de Planta/metabolismo , Vitis/metabolismo , Humanos , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/crescimento & desenvolvimento , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Vitis/efeitos dos fármacos , Vitis/crescimento & desenvolvimentoRESUMO
A novel methodology is presented for the enhanced electrochemical detection of oleuropein in complex plant matrices by Graphene Oxide Pencil Grahite Electrode (GOPGE) in combination with a buffer modified with a Natural Deep Eutectic Solvent, containing 10% (v/v) of Lactic acid, Glucose and H2 O (LGH). The electrochemical behavior of oleuropein in the modified-working buffer was examined using differential pulse voltammetry. The combination of both modifications, NADES modified buffer and nanomaterial modified electrode, LGH-GOPGE, resulted on a signal enhancement of 5.3 times higher than the bare electrode with unmodified buffer. A calibration curve of oleuropein was performed between 0.10 to 37 µM and a good linearity was obtained with a correlation coefficient of 0.989. Detection and quantification limits of the method were obtained as 30 and 102 nM, respectively. In addition, precision studies indicated that the voltammetric method was sufficiently repeatable, %RSD 0.01 and 3.16 (n = 5) for potential and intensity, respectively. Finally, the proposed electrochemical sensor was successfully applied to the determination of oleuropein in an olive leaf extract prepared by ultrasound-assisted extraction. The results obtained with the proposed electrochemical sensor were compared with Capillary Zone Electrophoresis analysis with satisfactory results.
Assuntos
Técnicas Eletroquímicas/métodos , Grafite/química , Iridoides/análise , Óxidos/química , Solventes/química , Técnicas Biossensoriais/métodos , Eletrodos , Eletroforese Capilar/métodos , Glucosídeos Iridoides , Iridoides/isolamento & purificação , Limite de Detecção , Olea/química , Extratos Vegetais/química , Folhas de Planta/químicaRESUMO
An approach to understand vines (Vitis vinifera) defense mechanism against heavy metal stress by isolation and determination of Hg-phytochelatins (PCs) complexes was performed. PCs are important molecules involved in the control of metal concentration in plants. PCs complex toxic metals through -SH groups and stores them inside cells vacuole avoiding any toxic effect of free metals in the cytosol. The Hg-PCs identification was achieved by determination of Hg and S as hetero-tagged atoms. A method involving two-dimensional chromatographic analysis coupled to atomic spectrometry and confirmation by tandem mass spectrometry is proposed. An approach involving size exclusion chromatography coupled to inductively coupled plasma mass spectrometry on roots, stems, and leaves extracts describing Hg distribution according to molecular weight and sulfur associations is proposed for the first time. Medium-low molecular weight Hg-S associations of 29-100 kDa were found, suggesting PCs presence. A second approach employing reversed-phase chromatography coupled to atomic fluorescence spectrometry analysis allowed the determination of Hg-PCs complexes within the mentioned fractions. Chromatograms showed Hg-PC2, Hg-PC3 and Hg-PC4 presence only in roots. Hg-PCs presence in roots was confirmed by ESI-MS/MS analysis.
Assuntos
Complexos de Coordenação/metabolismo , Mercúrio/metabolismo , Fitoquelatinas/metabolismo , Vitis/metabolismo , Adaptação Fisiológica , Cromatografia Líquida de Alta Pressão , Cromatografia de Fase Reversa , Complexos de Coordenação/isolamento & purificação , Mercúrio/isolamento & purificação , Fitoquelatinas/isolamento & purificação , Folhas de Planta/metabolismo , Raízes de Plantas/metabolismo , Caules de Planta/metabolismo , Estresse FisiológicoRESUMO
Concerns regard watering crops with Hg contaminated waters have arisen worldwide recently. In these sense Hg uptake by Vitis vinifera L. cv. Malbec was evaluated under greenhouse conditions by the administration of Hg(2+) for 4 days through irrigation water (short-term administration). Vines uptake Hg translocating it from roots through stems to leaves. Roots accumulated the higher Hg concentration. Hg in stems and leaves was accumulated mostly as organic Hg, bind to different moieties. Size exclusion chromatography (SEC) and ion pair chromatography (IPC) were employed to reach insights into these ligands. Hg is distributed mainly in high molecular weight fractions of 669 kDa in vine plants. In stems and leaves, Hg-S associations were found in 669 and 66 kDa fractions. Hg-S association at 66 kDa suggests a possible protein or peptide binding affecting vines normal physiology. Since Hg contamination through organomercurials is more harmful than Hg(2+) itself, methyl mercury, dimethyl mercury, and phenyl mercury, more toxic Hg species were evaluated with negative results.
Assuntos
Irrigação Agrícola , Mercúrio/análise , Medição de Risco/métodos , Vitis/metabolismo , Cromatografia em Gel , Mercúrio/toxicidade , Vitis/efeitos dos fármacosRESUMO
In this work, microwave-induced plasma optical emission spectrometry was applied for multielement determination in South American wine samples. The analytes were determined after acid digestion of 47 samples of Brazilian and Argentinian wines. Then, logistic regression, support vector machine, and decision tree for exploratory analysis and comparison of these algorithms in differentiating red wine samples by region of origin were carried out. All wine samples were classified according to their geographical origin. The quantification limits (mg L-1) were P: 0.06, B: 0.08, K: 0.17, Mn: 0.002, Cr: 0.002, and Al: 0.02. The accuracy of the method was evaluated by analyzing the wine samples by ICP OES for results' comparison. The concentrations in mg L-1 found for each element in wine samples were as follows: Al (< 0.02-1.82), Cr (0.15-0.50), Mn (< 0.002-0.8), P (97-277), B (1.7-11.6), Pb (< 0.06-0.3), Na (8.84-41.57), and K (604-1701), in mg L-1.
Assuntos
Oligoelementos , Vinho , Vinho/análise , Espectrometria de Massas/métodos , Inteligência Artificial , Análise Espectral/métodos , Brasil , Oligoelementos/análiseRESUMO
A high performance liquid chromatography coupled with atomic fluorescence spectrometry method for the determination of thimerosal (sodium ethylmercury thiosalicylate, C9H9HgNaO2S), ethylmercury, and inorganic mercury is proposed. Mercury vapor is generated by the post-column reduction of mercury species in formic acid media using UV-radiation. Thimerosal is quantitatively converted to Hg(II) followed by the reduction of Hg(II) to Hg(0). This method is applied to the determination of thimerosal (THM), ethylmercury (EtHg) and inorganic Hg in samples of a pharmaceutical industry effluent, and in waters of the San Luis River situated in the west side of San Luis city (Middle West, Argentine) where the effluents are dumped. The limit of detections, calculated on the basis of the 3σ criterion, where 0.09, 0.09 and 0.07 µg L(-1) for THM, EtHg(II) and for Hg(II), respectively. Linearity was attained from levels close to the detection limit up to at least 100 µg L(-1).
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Compostos de Etilmercúrio/análise , Espectrometria de Fluorescência/métodos , Timerosal/análise , Indústria Farmacêutica , Limite de Detecção , Mercúrio/análise , Rios , Espectrofotometria Atômica/métodosRESUMO
A simple non-chromatographic method for the determination of mercury (Hg(2+)), methylmercury (MeHg(+)), dimethylmercury (Me(2)Hg), and phenylmercury (PhHg(+)) employing atomic fluorescence spectrometry (AFS) as detection technique was developed. Mercury species showed a particular behavior in the presence of several reagents. In a first stage SnCl(2) was employed for Hg(2+) determination; in a second step, [Hg(2+)+PhHg(+)] concentration was determined using SnCl(2) and UV radiation. MeHg(+) decomposition was prevented adding 2-mercaptoethanol. In a third stage, [Hg(2+)+PhHg(+)+MeHg(+)] concentration was determined using K(2)S(2)O(8). Finally, the four species were determined employing NaBH(4). Reagents concentration and flow rates were optimized. The extraction technique of mercury species involved the use of 2-mercaptoethanol as ion-pair reagent. The limits of detection for Hg(2+), PhHg(+), MeHg(+), and Me(2)Hg were 1, 40, 68, and 99 ng L(-1) with a relative standard deviation of 1.5, 3.1, 4.7 and 5.8%, respectively. Calibration curve was linear with a correlation factor equal to 0.9995. The method was successfully applied to the determination of the mercury species in two Antarctic materials: IRMM 813 (Adamussium colbecki) and MURST-ISS-A2 (Antarctic Krill).