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Waste from the olive industry is a noticeable source of antioxidant compounds that can be extracted and reused to produce raw materials related to the chemical, cosmetic, food and pharmaceutical sectors. This work studies the phenolic composition of olive leaf samples using liquid chromatography with ultraviolet detection coupled to mass spectrometry (LC-UV-MS). Olive leaf waste samples have been crushed, homogenized, and subjected to a solid-liquid extraction treatment with mechanical shaking at 80 °C for 2 h using Natural Deep Eutectic Solvents (NaDES). The phenolic compound identification in the resulting extracts has been carried out by high-resolution mass spectrometry (HRMS) using data-dependent acquisition mode using an Orbitrap HRMS instrument. >60 different phenolic compounds have been annotated tentatively, of which about 20 have been confirmed from the corresponding standards. Some of the most noticeable compounds are oleuropein and its aglycone and glucoside form, luteolin-7-O-glucoside, 3-hydroxytyrosol, and verbascoside.
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Olea , Fenóis , Extratos Vegetais , Folhas de Planta , Olea/química , Folhas de Planta/química , Fenóis/química , Fenóis/análise , Extratos Vegetais/química , Espectrometria de Massas , Cromatografia Líquida de Alta PressãoRESUMO
A confirmatory method for the determination of polypeptide antibiotics (bacitracin, colistin, and polymyxin B) in muscle samples has been developed. Extraction is performed with acidified methanol, and a clean-up step by solid-phase extraction with polymeric cartridges is applied. Separation by ultra-high performance liquid chromatography (UHPLC) is carried out using a solid core C18 column and gradient elution with water/acetonitrile containing 0.2% formic acid. High-resolution mass spectrometry (HRMS) (Q-Orbitrap) detection using different working modes has proved to be highly advantageous in eliminating interfering signals from endogenous matrix components. The analytical method has been successfully validated according to Commission Regulation 2021/808/EU and is currently used in a public health laboratory involved in veterinary medicines residue surveillance activities.
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Antibacterianos , Espectrometria de Massas em Tandem , Animais , Antibacterianos/análise , Espectrometria de Massas em Tandem/métodos , Cromatografia Líquida de Alta Pressão/métodos , Músculos/química , Peptídeos , Extração em Fase Sólida/métodosRESUMO
Agrifood industries generate large amounts of waste that may result in remarkable environmental problems, such as soil and water contamination. Therefore, proper waste management and treatment have become an environmental, economic, and social challenge. Most of these wastes are exceptionally rich in bioactive compounds (e.g., polyphenols) with potential applications in the food, cosmetic, and pharmaceutical industries. Indeed, the recovery of polyphenols from agrifood waste is an example of circular bioeconomy, which contributes to the valorization of waste while providing solutions to environmental problems. In this context, unconventional extraction techniques at the industrial scale, such as microwave-assisted extraction (MAE), which has demonstrated its efficacy at the laboratory level for analytical purposes, have been suggested to search for more efficient recovery procedures. On the other hand, natural deep eutectic solvents (NADES) have been proposed as an efficient and green alternative to typical extraction solvents. This review aims to provide comprehensive insights regarding the extraction of phenolic compounds from agrifood waste. Specifically, it focuses on the utilization of MAE in conjunction with NADES. Moreover, this review delves into the possibilities of recycling and reusing NADES for a more sustainable and cost-efficient industrial application. The results obtained with the MAE-NADES approach show its high extraction efficiency while contributing to green practices in the field of natural product extraction. However, further research is necessary to improve our understanding of these extraction strategies, optimize product yields, and reduce overall costs, to facilitate the scaling-up.
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Agri-food industries generate a large amount of waste that offers great revalorization opportunities within the circular economy framework. In recent years, new methodologies for the extraction of compounds with more eco-friendly solvents have been developed, such as the case of natural deep eutectic solvents (NADES). In this study, a methodology for extracting phenolic compounds from olive tree leaves using NADES has been optimized. The conditions established as the optimal rely on a solvent composed of choline chloride and glycerol at a molar ratio of 1:5 with 30% water. The extraction was carried out at 80 °C for 2 h with constant agitation. The extracts obtained have been analyzed by high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) in MRM mode. The comparison with conventional ethanol/water extraction has shown that NADES, a more environmentally friendly alternative, has improved extraction efficiency. The main polyphenols identified in the NADES extract were Luteolin-7-O-glucoside, Oleuropein, 3-Hydroxytyrosol, Rutin, and Luteolin at the concentrations of 262, 173, 129, 34, and 29 mg kg-1 fresh weight, respectively.
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Winery wastes are rich in polyphenols with high added value to be used in cosmetics, pharmaceuticals, and food products. This work aims at recovering and purifying the polyphenolic fraction occurring in the malolactic fermentation lees generated during the production of Albariño wines. Phenolic acids, flavonoids, and related compounds were recovered from this oenological waste by green liquid extraction using water as the solvent. The resulting extract solution was microfiltered to remove microparticles and further treated by ultrafiltration (UF) using membranes of 30 kDa and 5 kDa molecular weight cut-offs (MWCOs). The feed sample and the filtrate and retentate solutions from each membrane system were analyzed by reversed-phase liquid chromatography (HPLC) with UV and mass spectrometric (MS) detection. The most abundant polyphenols in the extracts were identified and quantified, namely: caftaric acid with a concentration of 200 µg g-1 and trans-coutaric acid, cis-coutaric acid, gallic acid, and astilbin with concentrations between 15 and 40 µg g-1. Other minor phenolic acids and flavanols were also found. The UF process using the 30 kDa membrane did not modify the extract composition, but filtration through the 5 kDa poly-acrylonitrile membrane elicited a decrease in polyphenolic content. Hence, the 30 kDa membrane was recommended to further pre-process the extracts. The combined extraction and purification process presented here is environmentally friendly and demonstrates that malolactic fermentation lees of Albariño wines are a valuable source of phenolic compounds, especially phenolic acids.
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Polifenóis , Ultrafiltração , Polifenóis/análise , Extratos VegetaisRESUMO
Spinach and orange by-products are well recognized for their health benefits due to the presence of natural polyphenols with antioxidant activity. Therefore, the demand to produce functional products containing polyphenols recovered from vegetables and fruits has increased in the last decade. This work aims to use the integrated membrane process for the recovery of polyphenols from spinach and orange wastes, implemented on a laboratory scale. The clarification (microfiltration and ultrafiltration, i.e., MF and UF), pre-concentration (nanofiltration, NF), and concentration (reverse osmosis, RO) of the spinach and orange extracts were performed using membrane technology. Membrane experiments were carried out by collecting 1 mL of the permeate stream after increasing the flow rate in 1 mL/min steps. The separation and concentration factors were determined by HPLC-DAD in terms of total polyphenol content and by polyphenol families: hydroxybenzoic acids, hydroxycinnamic acids, and flavonoids. The results show that the transmembrane flux depended on the feed flow rate for MF, UF, NF, and RO techniques. For the spinach and orange matrices, MF (0.22 µm) could be used to remove suspended solids; UF membranes (30 kDa) for clarification; NF membranes (TFCS) to pre-concentrate; and RO membranes (XLE for spinach and BW30 for orange) to concentrate. A treatment sequence is proposed for the two extracts using a selective membrane train (UF, NF, and RO) to obtain polyphenol-rich streams for food, pharmaceutical, and cosmetic applications, and also to recover clean water streams.
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The aim of this study was to evaluate the recovery of phenolic compounds from olive mill and winery wastes by conventional solid-liquid extraction (SLE) using water as the extraction solvent. The studied variables were extraction time (5-15 min), temperature (25-90 °C), solid-to-liquid ratio (1:10-1:100 (kg/L)), pH (3-10) and application of multiple extractions (1-3). The extraction efficiency was evaluated in terms of total phenolic content (TPC), determined by high performance liquid chromatography (HPLC-UV), but also from the recovery of some representative phenolic compounds. The optimized conditions were one extraction step, 10 min, 25 °C, 1:30 (kg/L), pH 5 for olive pomace, and one extraction step, 10 min, 70 °C, 1:100 (kg/L), pH 5 for winery residues. The extraction method is simple and suitable for scaling-up in industry, and the aqueous extracts are fully compatible with further purification schemes based on the use of membranes or resins. The optimized technique was applied to a set of different representative residues from olive mill and winery industries, to assess their suitability as sources for phenolic compounds recovery. The phenolic content in the extracts was evaluated by chromatographic analysis and by the Folin-Ciocalteu assay (FC). Furthermore, the antioxidant capacity was determined by 2,2-azinobis-3-etilbenzotiazolina-6-sulfonat (ABTS), 2,-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays. Because of their high contents in phenolic compounds and great antioxidant capacity, olive pomace and lees filters were identified as especially suited sources for phenolic compounds recovery.
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Olea , Antioxidantes/química , Cromatografia Líquida de Alta Pressão , Olea/química , Fenóis/química , Solventes/química , ÁguaRESUMO
More sustainable waste management in the winery and olive oil industries has become a major challenge. Therefore, waste valorization to obtain value-added products (e.g., polyphenols) is an efficient alternative that contributes to circular approaches and sustainable environmental protection. In this work, an integration scheme was purposed based on sustainable extraction and membrane separation processes, such as nanofiltration (NF) and reverse osmosis (RO), for the recovery of polyphenols from winery and olive mill wastes. Membrane processes were evaluated in a closed-loop system and with a flat-sheet membrane configuration (NF270, NF90, and Duracid as NF membranes, and BW30LE as RO membrane). The separation and concentration efficiency were evaluated in terms of the total polyphenol content (TPC), and by polyphenol families (hydroxybenzoic acids, hydroxycinnamic acids, and flavonoids), using high-performance liquid chromatography. The water trans-membrane flux was dependent on the trans-membrane pressure for the NF and RO processes. NF90 membrane rejected around 91% of TPC for the lees filters extracts while NF270 membrane rejected about 99% of TPC for the olive pomace extracts. Otherwise, RO membranes rejected more than 99.9% of TPC for both types of agri-food wastes. Hence, NF and RO techniques could be used to obtain polyphenol-rich streams, and clean water for reuse purposes.
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Total polyphenol content and antioxidant capacity were estimated in various food and nutraceutical samples, including cranberries, raspberries, artichokes, grapevines, green tea, coffee, turmeric, and other medicinal plant extracts. Samples were analyzed by using two antioxidant assays-ferric reducing antioxidant power (FRAP) and Folin-Ciocalteu (FC)-and a reversed-phase high-performance liquid chromatography (HPLC), with a focus on providing compositional fingerprints dealing with polyphenolic compounds. A preliminary data exploration via principal component analysis (PCA) revealed that HPLC fingerprints were suitable chemical descriptors to classify the analyzed samples according to their nature. Moreover, chromatographic data were correlated with antioxidant data using partial least squares (PLS) regression. Regression models have shown good prediction capacities in estimating the antioxidant activity from chromatographic data, with determination coefficients (R2) of 0.971 and 0.983 for FRAP and FC assays, respectively.
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The production of olive oil and wine are two of the main agri-food economic activities in Southern Europe. They generate large amounts of solid and liquid wastes (e.g., olive pomace, olive mill wastewater, grape pomace, grape stems, wine lees, and wine processing wastewater) that represent a major environmental problem. Consequently, the management of these residues has become a big challenge for these industries, since they are harmful to the environment but rich in bioactive compounds, such as polyphenols. In recent years, the recovery of phenolic compounds has been proposed as a smart strategy for the valorization of these by-products, from a circular economy perspective. This review aims to provide a comprehensive description of the state of the art of techniques available for the analysis, extraction, and purification of polyphenols from the olive mill and winery residues. Thus, the integration and implementation of these techniques could provide a sustainable solution to the olive oil and winery sectors.
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Phenolic compounds recovery by mechanical stirring extraction (MSE) was studied from orange and spinach wastes using water as a solvent. The statistical analysis showed that the highest total polyphenol content (TPC) yield was obtained using 15 min, 70 °C, 1:100 (w/v) solid/solvent ratio and pH 4 for orange; and 5 min, 50 °C, 1:50 (w/v) solid/solvent ratio and pH 6 for spinach. Under these conditions, the TPC was 1 mg gallic acid equivalent (GAE) g-1 fresh weight (fw) and 0.8 mg GAE g-1 fw for orange and spinach, respectively. MSE substantially increased the phenolic compounds yields (1-fold for orange and 2-fold for spinach) compared with ultrasound-assisted extraction. Furthermore, the antioxidant activity of orange and spinach extracts was evaluated using DPPH, FRAP and ABTS. The obtained results pointed out that the evaluated orange and spinach residues provided extracts with antioxidant activity (2.27 mg TE g-1 and 0.04 mg TE g-1, respectively).
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Fruits, vegetables, spices, and herbs are a potential source of phenolic acids and polyphenols. These compounds are known as natural by-products or secondary metabolites of plants, which are present in the daily diet and provide important benefits to the human body such as antioxidant, anti-inflammatory, anticancer, anti-allergic, antihypertensive and antiviral properties, among others. Plentiful evidence has been provided on the great potential of polyphenols against different viruses that cause widespread health problems. As a result, this review focuses on the potential antiviral properties of some polyphenols and their action mechanism against various types of viruses such as coronaviruses, influenza, herpes simplex, dengue fever, and rotavirus, among others. Also, it is important to highlight the relationship between antiviral and antioxidant activities that can contribute to the protection of cells and tissues of the human body. The wide variety of action mechanisms of antiviral agents, such as polyphenols, against viral infections could be applied as a treatment or prevention strategy; but at the same time, antiviral polyphenols could be used to produce natural antiviral drugs. A recent example of an antiviral polyphenol application deals with the use of hesperidin extracted from Citrus sinensis. The action mechanism of hesperidin relies on its binding to the key entry or spike protein of SARS-CoV-2. Finally, the extraction, purification and recovery of polyphenols with potential antiviral activity, which are essential for virus replication and infection without side-effects, have been critically reviewed.
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COVID-19 , Viroses , Antioxidantes , Antivirais , Humanos , Polifenóis , SARS-CoV-2 , Viroses/tratamento farmacológico , Viroses/prevenção & controleRESUMO
In this study, the recovery of polyphenols from olive oil mill and winery waste was investigated. The performance of ultrasound assisted extraction (UAE), microwave assisted extraction (MAE), and pressurized liquid extraction (PLE) was assessed using ethanol-water mixtures, which are compatible with food, nutraceutical, and cosmetic applications. The extraction efficiency from olive pomace and lees samples was evaluated in terms of total polyphenol content (TPC), determined by high performance liquid chromatography (HPLC) and Folin-Ciocalteu assay. The effect of solvent composition, temperature, and time was analyzed by response surface methodology. Ethanol:water 50:50 (v/v) was found to be a suitable solvent mixture for both kinds of samples and all three extraction techniques. The performance of the extraction techniques was evaluated, under optimal experimental conditions, with a set of different representative samples of residues from olive oil and wine production. Overall, the best extraction efficiency for olive pomace residues was provided by MAE (ethanol:water 50:50 (v/v), 90 °C, 5 min), and for wine residues by PLE (ethanol:water 50:50 (v/v), 100 °C, 5 min, 1 cycle). However, the results provided by UAE (ethanol:water 50:50 (v/v), 30 min) were also suitable. Considering not only extraction performance, but also investment and operational costs, UAE is proposed for a future scaling up evaluation. Regarding olive pomace as a source for natural phenolic antioxidants, olive variety and climatic conditions should be taken into account, since both influence TPC in the extracts, while for winery residues, lees from red wines are more suitable than those from white wines.
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This paper evaluates the antioxidant ability of polyphenols as a function of their chemical structures. Several common food indexes including Folin-Ciocalteau (FC), ferric reducing antioxidant power (FRAP) and trolox equivalent antioxidant capacity (TEAC) assays were applied to selected polyphenols that differ in the number and position of hydroxyl groups. Voltammetric assays with screen-printed carbon electrodes were also recorded in the range of -0.2 to 0.9 V (vs. Ag/AgCl reference electrode) to investigate the oxidation behavior of these substances. Poor correlations among assays were obtained, meaning that the behavior of each compound varies in response to the different methods. However, we undertook a comprehensive study based on principal component analysis that evidenced clear patterns relating the structures of several compounds and their antioxidant activities.
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A rapid and sensitive method for the confirmatory analysis of eight synthetic corticosteroids (betamethasone, dexamethasone, prednisolone, 6-methylprednisolone, triamcinolone, flumethasone, beclomethasone, fluocinolone acetonide) is proposed. The method is useful for detecting illegal treatments in different animal species. It consists of an extraction and cleanup using the quick, easy, cheap, effective, rugged, and safe (QuEChERS) strategy. Quantitative determination is achieved by ultrahigh-performance liquid chromatography coupled to high-resolution mass spectrometry with heated electrospray ionization in negative mode. Quantification is performed using surrogate matrix-matched standard calibration curve with dexamethasone-D4 as the internal standard. The method was validated for analyzing liver samples according to the criteria established by Decision 2002/657/EC. Linearity was assessed in the 1-10 µg kg-1 range and linear correlation coefficients were over 0.99 for all the analytes. CCα ranged from 0.04 to 0.16 µg kg-1 for substances without maximum residue limit. The method allows confident quantification and confirmation of corticosteroids in liver samples, and its simplicity makes it suitable for analyzing large numbers of samples.
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Corticosteroides/química , Cromatografia Líquida/métodos , Espectrometria de Massas/métodos , Animais , Estrutura Molecular , Sensibilidade e Especificidade , Fatores de TempoRESUMO
We developed a Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) method for the high throughput determination of 10 non-steroidal anti-inflammatory drugs (NSAIDs) in milk samples using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) with a triple quadrupole (QqQ) instrument and an electrospray ionization (ESI) source. The new extraction procedure is highly efficient, and we obtained absolute recoveries in the range 78.1-97.1 % for the extraction and clean-up steps. Chromatographic separation is performed in the gradient mode with a biphenyl column and acidic mobile phases consisting of water and acetonitrile containing formic acid. The chromatographic run time was about 12 min, and NSAID peaks showed a good symmetry factor. For MS/MS detection, we used multiple reaction monitoring (MRM) mode, using ESI in both positive and negative modes. Our method has been validated in compliance with the European Commission Decision 657/2002/EC, and we obtained very satisfactory results in inter-laboratory testing. Furthermore, we explored the use of a hybrid high resolution mass spectrometer, combining a quadrupole and an Orbitrap mass analyzer, for high resolution (HR) MS/MS detection of NSAIDs. We achieved lower NSAID quantification limits with Q-Orbitrap high resolution mass spectrometry (HRMS/MS) detection than those achieved with the QqQ instrument; however, its main feature is its very high selectivity, which makes HRMS/MS particularly suitable for confirmatory analysis.
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Anti-Inflamatórios não Esteroides/análise , Cromatografia Líquida/métodos , Leite/química , Espectrometria de Massas em Tandem/métodos , Animais , Anti-Inflamatórios não Esteroides/isolamento & purificação , Análise de Alimentos/métodos , Contaminação de Alimentos/análise , Limite de DetecçãoRESUMO
Sensitive and unequivocal determination of analytes/contaminants in complex matrices is a challenge in the field of food safety control. In this study, various acquisition modes (Full MS/AIF, Full MS+tMS/MS, Full MS/dd MS/MS and tSIM/ddMS/MS) and parameters of a quadrupole-orbitrap hybrid mass spectrometer (Q Exactive) were studied in detail. One of the main conclusions has been that, reducing the scan range for Full MS (using the quadrupole) and targeted modes give higher signal-to-noise (S/N) ratios and thereby better detection limits for analytes in matrix. The use of Q Exactive in a complex case, for the confirmatory analysis of hormones in animal urine is presented. A targeted SIM data dependent MS/MS (tSIM/ddMS/MS) acquisition method for determination of eight synthetic hormones (trenbolone, 17α ethinylestradiol, zeranol, stanozolol, dienestrol, diethylstilbestrol, hexestrol, taleranol) and a naturally occurring hormone (zearalenone) in animal urine were optimized to have sensitive precursors from targeted SIM mode and trigger MS/MS scans over the entire chromatograph peak. The method was validated according to EC/657/2002. CCα (decision limit) for the analytes ranged between 0.11 µg L(-1) and 0.69 µg L(-1) and CCß (detection capability) ranged between 0.29 µg L(-1) and 0.90 µg L(-1).
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Bovinos/urina , Hormônios/química , Hormônios/urina , Espectrometria de Massas/instrumentação , Animais , Espectrometria de Massas/métodosRESUMO
Commercial hardwood biochars ranging in N2 specific surface area of 0.1-427 m(2) · g(-1) were added to an agricultural soil at 0, 1, or 2% levels to determine whether they would predictably reduce the pore water concentration of sulfamethazine (SMT). The soil and biochar-soil mixtures were preweathered under mild (2 d, 20 °C) or more severe (28 d, 40 °C) conditions before spiking. The carbon-normalized biochar-water distribution coefficient (KBC) of the biochars varied by a factor of up to 10(4), depending on biochar properties and SMT concentration. Except for the fast-pyrolysis biochar, KBC greatly exceeded the soil organic carbon-water distribution coefficient KOC. Sorption in the mixtures increased as expected with biochar and dose. However, sorption was dramatically overpredicted (by up to 10(2.5)) by the sum of sorption to the individual components, indicating a strong weathering effect even under the mild conditions. The soil-subtracted weathered biochar-water isotherms were more linear, and the KBC values approached or lay within the range of KOC values reported for SMT in 19 soils. Biochars both in intimate contact with soil and placed in a membrane bag suspended in the solution showed reduced N2-B.E.T. surface area after weathering, implicating fouling of the biochar surface by humic substances transferred through water. The results indicate that only highly surfaceous, carbonaceous biochars would be useful for stabilizing soil contaminated with compounds such as SMT. They also suggest that weathering may attenuate the contribution of native (environmental) black carbon to sorption of such compounds in soils and sediments.
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Anti-Infecciosos/química , Poluentes do Solo/química , Solo/química , Sulfametazina/química , AdsorçãoRESUMO
Adsorption of ionizable compounds by black carbon is poorly characterized. Adsorption of the veterinary antibiotic sulfamethazine (SMT; a.k.a., sulfadimidine; pK(a1) = 2.28, pK(a2) = 7.42) on a charcoal was determined as a function of concentration, pH, inorganic ions, and organic ions and molecules. SMT displayed unconventional adsorption behavior. Despite its hydrophilic nature (log K(ow) = 0.27), the distribution ratio K(d) at pH 5, where SMT(0) prevails, was as high as 10(6) L kg(-1), up to 10(4) times greater than literature reported K(oc). The K(d) decreases at high and low pH but not commensurate with the decline in K(ow) of the ionized forms. At pH 1, where SMT(+) is predominant and the surface is positive, a major driving force is π-π electron donor-acceptor interaction of the protonated aniline ring with the π-electron rich graphene surface, referred to as π(+)-π EDA, rather than ordinary electrostatic cation exchange. In the alkaline region, where SMT(-) prevails and the surface is negative, adsorption is accompanied by near-stoichiometric proton exchange with water, leading to the release of OH(-) and formation of an exceptionally strong H-bond between SMT(0) and a surface carboxylate or phenolate, classified as a negative charge-assisted H-bond, (-)CAHB. At pH 5, SMT(0) adsorption is accompanied by partial proton release and is competitive with trimethylphenylammonium ion, signifying contributions from SMT(+) and/or the zwitterion, SMT(±), which take advantage of π(+)-π EDA interaction and Coulombic attraction to deprotonated surface groups. In essence, both pK(a1) and pK(a2) increase, and SMT(±) is stabilized, in the adsorbed relative to the dissolved state.
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Antibacterianos/química , Carvão Vegetal/química , Sulfametazina/química , Concentração de Íons de Hidrogênio , Modelos QuímicosRESUMO
A liquid chromatography-mass spectrometry (LC-MS) method was developed for the determination of five macrolides in natural water samples, using kitasamycin as surrogate. The macrolides were extracted from water samples using Oasis HLB cartridges. Pre-concentration factors up to 250 were obtained. Separation was carried out in an end-capped silica-based C18 column and mobile phases consisting of water/acetonitrile mixtures containing ammonium acetate. Detection was performed by mass spectrometry with a single quadrupole and a triple quadrupole using an electrospray interface. The quality parameters obtained with these two approaches were compared. The detection limits of the whole process were about 1 ng l(-1). The recoveries from 250 ml of water samples spiked at 25-125 ng l(-1) level were in the range 85-115%, except for azithromycin levels, which were around 70%. Erythromycin-H2O, clarithromycin and azithromyzin were found, at the sub ng l(-1) level, in the studied rivers.