Evaluating the catalytic effect of Fe@Fe2O3-modified granulated cork as an innovative heterogeneous catalyst in electro-fenton degradation of benzoquinone in different aqueous matrices.

This study investigated the potential of a novel biomass-derived cork as a suitable catalyst after its modification with Fe@Fe2O3 for in-situ application in heterogeneous electro-Fenton (HEF) process for benzoquinone (BQ) elimination from water. No attempts on the application of modified granulated cork (GC) as a suspended heterogeneous catalyst in the HEF process for water treatment have been published yet. GC was modified by sonification approach in a FeCl3 + NaBH4 solution to reduce the ferric ions to metallic iron in order to obtain Fe@Fe2O3-modified GC (Fe@Fe2O3/GC). Results clearly demonstrated that this catalyst exhibited excellent electrocatalytic properties, such as a high conductivity as well as relatively high redox current and possessed several active sites for water depollution applications. Using Fe@Fe2O3/GC as catalyst in HEF, 100% of BQ removal was achieved in synthetic solutions by applying 33.3 mA cm-2 after 120 min. Different experimental conditions were tested to determine that best possible conditions can be as follow: 50 mmol L-1 Na2SO4 and 10 mg L-1 of Fe@Fe2O3/GC catalyst using Pt/carbon-PTFE air diffusion cell by applying 33.3 mA cm-2. Nevertheless, when Fe@Fe2O3/GC was used in the HEF approach to depollute real water matrices, no complete BQ concentration was removal achieved after 300 min of treatment, achieving between 80 and 95% of effectiveness.

Combined effects of the vessel type and bottle closure during Chilean Sauvignon Blanc wine storage over its volatile profile.

A Sauvignon Blanc wine was subjected to a maturation period of six months by using four different types of vessels in triplicate: cylindrical stainless steel tanks, oval-shaped polyethylene tanks, cubic-shaped polyethylene tanks, and clay jars. After maturation in the different vessels, wines were bottled using three different closures (natural cork, synthetic cork, and screwcaps). The volatile compound profiles of the wine samples were recorded by SPME-GC-MS throughout vessel maturation as well as after the bottle storage period. In general terms, wines stored in stainless steel tanks showed the highest contents of volatile compounds when compared with the other tested vessels. Moreover, wines from bottles capped with screwcaps showed the highest contents of most of the volatile compounds when compared with the other closures. Moreover, an interaction between the vessel and the closure was observed: when screwcaps were used during bottle aging, the resulting wines were very similar to those matured in stainless steel vessels. These results suggest that the use of screwcaps hides the differences originating from wine composition during maturation in vessels other than stainless steel.

An Electroanalytical Solution for the Determination of Pb2+ in Progressive Hair Dyes Using the Cork-Graphite Sensor.

Lead is one of the most toxic metals for living organisms: once absorbed by soft tissues, it is capable of triggering various pathologies, subsequently bioaccumulating in the bones. In consideration of this, its detection and quantification in products for human consumption and use is of great interest, especially if the procedure can be carried out in an easy, reproducible and economical way. This work presents the results of the electroanalytical determination of lead in three different commercial products used as progressive hair dyes. Analyses were performed by cyclic voltammetry (CV) and differential pulse stripping voltammetry (DPSV) using a composite cork-graphite sensor in 0.5M H2SO4 solution or 0.1M acetate buffer (pH 4.5), in the presence and absence of hair dye samples. The H2SO4 solution gave better results in terms of analyte sensitivity than the acetate buffer electrolyte. In both cases, well-defined signals for lead were obtained by DPSV analyses, enabling the calibration curve and figures of merit to be determined. The limits of detection (LOD) were found to be approximately 1.06 µM and 1.26 µM in H2SO4 and acetate buffer, respectively. The DPSV standard addition method was successfully applied to quantify the lead in hair dye samples, yielding values below 0.45% in Pb. All three analyzed samples were shown to comply with the limit set by the Brazilian Health Regulatory Agency, i.e., 0.6% lead in this type of product. The comparison of the electroanalytical results with those obtained by the reference method, based on the use of inductively coupled plasma optical emission spectrometry (ICP-OES), confirmed that the electroanalytical detection approach is potentially applicable as a strategy for quality control.

Elemental characterization of sparkling wine and cork stoppers.

The variations of the elemental concentrations in sparkling white wine and respective cork stoppers throughout 18 months of storage time were determined with the PIXE (Particle-Induced X-ray Emission) technique. Three portions of the cork stoppers were analyzed: the top part (external layer), the inner part (bulk layer) and the bottom layer (which was in contact with the sparkling wine). Elements such as Na, Mg, Si, P, S, Cl, K, Ca, Mn, Fe, Zn and Rb were determined for both cork stoppers and sparkling wine samples. Similar concentrations of Si, P, S, Cl and Ca were found in the external and bottom layers of the corks. Distinct behaviors of the changes in the elemental concentrations as a function of the time were observed for cork stoppers and sparkling wines. The concentrations of Mg, S, K, Ca, Cu, Sr and Ba increased in the bottom layer of the cork as a function of storage time. On the other hand, concentrations of Al, Si, Cl, Ti, Zn and Br proved to be invariant, while the concentrations of P and Fe showed a slight decrease. Concerning the sparkling wine, an increasing trend of elemental concentrations was observed for most elements throughout the storage time. A diffusion mechanism of elements in the cork and the role of the secondary fermentation in the bottle are discussed.

Green Composite Sensor for Monitoring Hydroxychloroquine in Different Water Matrix.

Hydroxychloroquine (HCQ), a derivative of 4-aminoquinolone, is prescribed as an antimalarial prevention drug and to treat diseases such as rheumatoid arthritis, and systemic lupus erythematosus. Recently, Coronavirus (COVID-19) treatment was authorized by national and international medical organizations by chloroquine and hydroxychloroquine in certain hospitalized patients. However, it is considered as an unproven hypothesis for treating COVID-19 which even itself must be investigated. Consequently, the high risk of natural water contamination due to the large production and utilization of HCQ is a key issue to overcome urgently. In fact, in Brazil, the COVID-19 kit (hydroxychloroquine and/or ivermectin) has been indicated as pre-treatment, and consequently, several people have used these drugs, for longer periods, converting them in emerging water pollutants when these are excreted and released to aquatic environments. For this reason, the development of tools for monitoring HCQ concentration in water and the treatment of polluted effluents is needed to minimize its hazardous effects. Then, in this study, an electrochemical measuring device for its environmental application on HCQ control was developed. A raw cork-graphite electrochemical sensor was prepared and a simple differential pulse voltammetric (DPV) method was used for the quantitative determination of HCQ. Results indicated that the electrochemical device exhibited a clear current response, allowing one to quantify the analyte in the 5-65 µM range. The effectiveness of the electrochemical sensor was tested in different water matrices (in synthetic and real) and lower HCQ concentrations were detected. When comparing electrochemical determinations and spectrophotometric measurements, no significant differences were observed (mean accuracy 3.0%), highlighting the potential use of this sensor in different environmental applications.

Applicability of Cork as Novel Modifiers to Develop Electrochemical Sensor for Caffeine Determination.

This study aims to investigate the applicability of a hybrid electrochemical sensor composed of cork and graphite (Gr) for detecting caffeine in aqueous solutions. Raw cork (RAC) and regranulated cork (RGC, obtained by thermal treatment of RAC with steam at 380 °C) were tested as modifiers. The results clearly showed that the cork-graphite sensors, GrRAC and GrRGC, exhibited a linear response over a wide range of caffeine concentration (5-1000 µM), with R2 of 0.99 and 0.98, respectively. The limits of detection (LOD), estimated at 2.9 and 6.1 µM for GrRAC and GrRGC, suggest greater sensitivity and reproducibility than the unmodified conventional graphite sensor. The low-cost cork-graphite sensors were successfully applied in the determination of caffeine in soft drinks and pharmaceutical formulations, presenting well-defined current signals when analyzing real samples. When comparing electrochemical determinations and high performance liquid chromatography measurements, no significant differences were observed (mean accuracy 3.0%), highlighting the potential use of these sensors to determine caffeine in different samples.

Designing a green device to BAµE: Recycled cork pellet as extraction phase for the determination of parabens in river water samples.

This study reports a novel and environmentally friendly method based on bar adsorptive microextraction (BAµE) with cork pellet as extraction phase for the determination of methylparaben, ethylparaben, propylparaben and butylparaben in river water samples. This natural approach consists of a cork pellet recycled from wine stoppers used as biosorbent material to replace the traditional BAµE device. The analytical determinations were performed using a high-performance liquid chromatography-diode array detector (HPLC-DAD). Parameters such as type of desorption solvent, desorption and extraction time, sample pH and ionic strength were carefully optimized through univariate and multivariate approaches. Cork pellets of 15 mm length were inserted into vials containing 15 mL of water sample adjusted at pH 3 and 25% (w/v) of NaCl. The extraction step was carried out under agitation for 45 min followed by liquid desorption with 120 µL of methanol:acetonitrile (1:1 v/v) for 30 min. Satisfactory analytical performance was obtained with coefficients of determination ranging from 0.9921 for methylparaben to 0.9994 for propylparaben; intraday precision ranged from 6.7 to 18.3%, and interday precision varied from 7.2 to 20.0%. Accuracy was assessed through relative recovery assays and varied from 53 to 124%.

A high throughput approach to rotating-disk sorptive extraction (RDSE) using laminar cork for the simultaneous determination of multiclass organic micro-pollutants in aqueous sample by GC-MS.

In this study, a high throughput approach to rotating-disk sorptive extraction (RDSE) using laminar cork as extraction phase is demonstrated for the first time in the determination of 20 multiclass organic micro-pollutants including pesticides, PAHs and UV filters compounds from aqueous samples with gas chromatography mass spectrometry (GC-MS). The influencing parameters (desorption solvent, volume and time, extraction time and sample pH and ionic strength) were carefully optimized using multivariate designs. The optimal conditions were 10 min for extraction using 35 mL of water samples and a liquid desorption using 1 mL of MeOH:AcOEt (50:50% v/v) for 20 min. A low-cost apparatus that allows six extractions simultaneously, providing a high throughput of 5 min per sample turnaround times, considering the sample preparation step was used for the first time in this modified RDSE methodology. Satisfactory analytical performance was achieved with limits of detection (LOD) between 0.08 and 1.5 µg L-1 and limits of quantification (LOQ) between 0.3 and 4.8 µg L-1. The relative recoveries for the analytes were determined using river and lake water samples spiked at different concentrations and ranging from 80% to 119% for all analytes, with relative standard deviations (RSD) lower than 20%. The extraction efficiency obtained for the proposed configuration with laminar cork was significantly superior to powdered cork, demonstrating an interesting new configuration for new applications.

Cork sheet as a sorptive phase to extract hormones from water by rotating-disk sorptive extraction (RDSE).

This work reports for the first time the use of laminar cork as a sorptive phase in a microextraction technique, rotating-disk sorptive extraction (RDSE). Typical hormones (estrone, estradiol, estriol and ethinyl estradiol) were selected as analyte models and extracted from wastewater samples on laminar cork with statistically equivalent extraction efficiency to that provided by Oasis HLB. The cork characterization was performed by confocal fluorescence microscopy (CLSM), Fourier-transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM), allowing the identification of lignin, suberin and polysaccharides (cellulose and hemicellulose) as the main components of the cork. The best conditions for extraction were as follows: rotation velocity of the disk, 2000 rpm; extraction time, 45 min; and sample volume, 20 mL. The analytical features of the developed method show that calibration curves for all analytes have R2 values higher than 0.99. The absolute recoveries were higher than 63%, and the precision, expressed as relative standard deviation, ranged from 2 to 16%. The LOD and LOQ ranges were 3-19 and 10-62 ng L-1, respectively. The proposed method was applied to the analysis of wastewater, and the concentrations of hormones in a wastewater treatment plant in Santiago, Chile, ranged from

Impact of closure type and storage temperature on chemical and sensory composition of Malbec wines (Mendoza, Argentina) during aging in bottle.

Malbec is the flagship variety of Argentina mainly due to its high oenological value and plasticity to obtain different wine styles. During bottled aging, the chemical and organoleptic composition of wines is subject to changes depending on the aging conditions (closure, oxygen level, temperature, time). However, the combined effect of these factors on chemical composition and organoleptic characteristics of Malbec wines has not been studied yet. Wines were bottled with screw cap and natural cork and were kept in chambers at 15 °C and 25 °C for 2 years. Sampling was performed at 2, 4, 6, 9, 12, 15, 18, 21 and 24 months. Concentrations of free sulfur dioxide, dissolved oxygen, anthocyanins, tannins, esters, volatile phenols, organic acids, and color saturation decreased during the storage process. While, the formation of polymeric pigments, the color attributes (lightness and hue) and the levels of alcohols, norisoprenoids, furanoids and terpenoids increased. At 24 months, Malbec wines were organoleptically different. Wines kept at 15 °C were associated with high sensory perceptions in color intensity and violet tint, those presented a positive correlation with free sulfur dioxide, tannins, and anthocyanins levels. On the contrary, wines aged at 25 °C were linked with high sensory perceptions of dried vegetative and dried fruit aromas. These descriptors were positively correlated with norisoprenoids, furanoids, and terpenoids. In general, the chemical composition and organoleptic attributes of bottled Malbec wines (Mendoza, Argentina) were stable respect closure type employed, but highly sensitive to the combined effect of time and storage temperature. This finding is key to making decisions about the wine style searched, and costs (e.g. refrigeration) involved in the conservation period until consumption.

A natural and renewable biosorbent phase as a low-cost approach in disposable pipette extraction technique for the determination of emerging contaminants in lake water samples.

This study proposes an efficient analytical methodology using a biosorbent (cork) as an extraction phase in disposable pipette extraction technique for the rapid determination of the emerging contaminants methyl paraben, ethyl paraben, benzophenone, 3-(4-methylbenzylidene) camphor and 2-(ethylhexyl)-4-(dimethylamino) benzoate in lake water samples using high-performance liquid chromatography with diode array detection. The optimized conditions were comprised of 800 µL of sample, three cycles of 30 s each for the extraction, pH 6, addition of 30% w/v of NaCl. For the desorption step, the optimized desorption conditions were achieved with 100 µL of a mixture comprised of 50% methanol and 50% acetonitrile v/v, using one cycle of 30 s. Excellent analytical performance was achieved with limits of detection of 0.6 µg/L for methyl paraben to 1.4 µg/L for 3-(4-methylbenzylidene) camphor, and the limit of quantitation varied from 2 µg/L for methyl paraben to 4.3 µg/L 3-(4-methylbenzylidene) camphor, respectively. The correlation coefficients ranged from 0.9962 for ethyl paraben to 0.9980 for methyl paraben. The method accuracy varied from 71-132%, and the intraday precision ranged from 3 to 23% (n = 3) and interday from 9 to 23% (n = 9). The robustness was evaluated through Youden and Lenth's methods and indicated no significant variations in the results.

Expanding the applicability of cork as extraction phase for disposable pipette extraction in multiresidue analysis of pharmaceuticals in urine samples.

In this paper, cork is proposed as a natural and renewable material for the extraction phase in disposable pipette extraction to be applied in the multiresidue determination of pharmaceuticals in human urine by HPLC with diode array detection. The compounds carbamazepine, losartan, ketoprofen, 17­ß­estradiol, naproxen, diazepam, 17­α­ethinylestradiol, estrone, diclofenac and ibuprofen were studied. A known amount (5 mg) of cork, with a size of 200 mesh, was inserted into a pipette tip. The method optimization was carried out with univariate and multivariate approaches. The optimized conditions were sample pH adjusted to 3, urine dilution factor of 40 (158 µL of urine diluted in 6.142 mL of ultrapure water), 9 extraction cycles each performed with 700 µL of sample, and extraction time of 10 s per cycle. Desorption was performed with 85 µL of methanol applying 6 cycles of 10 s each using the same solvent aliquot. For the clean-up step, 4 cycles were carried out each with 200 µL of methanol. The limits of quantification varied from 5 to 10 µg L-1 with determination coefficients higher than 0.9919 for the calibration curves for all the analytes. Intra-day and inter-day precision and relative recovery from urine samples donated by two volunteers were assessed based on three spiked concentrations. The analyte relative recoveries ranged from 65 to 117% (n = 3) for the two samples. Intra-day precision ranged from 1.2 to 17% (n = 3) and inter-day precision varied from 11 to 21% (n = 9).

Exploiting green sorbents in rotating-disk sorptive extraction for the determination of parabens by high-performance liquid chromatography with tandem electrospray ionization triple quadrupole mass spectrometry.

In this study, the viability of applying cork and montmorillonite clay modified with ionic liquid as biosorbents in the rotating-disk sorptive extraction technique was investigated. Specifically, this was aimed at the determination of methyl paraben, ethyl paraben, propyl paraben, and isobutyl paraben, with separation/determination by high-performance liquid chromatography coupled with mass spectrometry. The optimization of the method for both biosorbents was performed using multivariate procedures. The extraction efficiencies for the target compounds in aqueous matrices were compared to those obtained using the commercial sorbent Octadecil Silano-C18. The optimum extraction conditions for both natural sorbents were the use of an ammonia solution (pH 10) as desorption solvent and an extraction time of 30 min. The proposed methods show limits of quantification of 0.8 µg/L for cork, 3.0 µg/L for montmorillonite clay and 6.0 µg/L for Octadecil Silano-C18. The relative recoveries from river water and tap water samples ranged from 80.3 to 118.7% and 80.0 to 119.9% for cork and montmorillonite clay modified with ionic liquid, respectively. Relative standard deviations were obtained for intra- and interday precisions of <15% and <19% for cork and <19% and <17% for montmorillonite clay modified with ionic liquid.

Development of a high-throughput method based on thin-film microextraction using a 96-well plate system with a cork coating for the extraction of emerging contaminants in river water samples.

In this study, a new method was developed in which a biosorbent material is used as the extractor phase in conjunction with a recently described sample preparation technique called thin-film microextraction and a 96-well plate system. The method was applied for the determination of emerging contaminants, such as 3-(4-methylbenzylidene) camphor, ethylparaben, triclocarban, and bisphenol A in water samples. The separation and detection of the analytes were performed by high-performance liquid chromatography with diode array detection. These contaminants are considered hazardous to human health and other living beings. Thus, the development of an analytical method to determine these compounds is of great interest. The extraction parameters were evaluated using multivariate and univariate optimization techniques. The optimum conditions for the method were 3 h of extraction time, 20 min of desorption with 300 µL of acetonitrile and methanol (50:50, v/v), and the addition of 5% w/v sodium chloride to the sample. The analytical figures of merit showed good results with linear correlation coefficients higher than 0.99, relative recoveries of 72-125%, interday precision (n = 3) of 4-18%, and intraday precision (n = 9) of 1-21%. The limit of detection was 0.3-5.5 µg/L, and the limit of quantification was 0.8-15 µg/L.

A new configuration for bar adsorptive microextraction (BAµE) for the quantification of biomarkers (hexanal and heptanal) in human urine by HPLC providing an alternative for early lung cancer diagnosis.

In this paper, a remodeling of the bar adsorptive microextraction (BAµE) technique is proposed with impregnation of the derivatization reagent on the surface of the adsorptive bar containing a biosorbent material. The derivatization reagent was 2,4-dinitrophenylhydrazine (DNPH), which was adsorbed on the surface of the bar containing cork powder as the extractor phase for the determination of two aldehydes (hexanal and heptanal) which are known as lung cancer biomarkers in human urine samples. The derivatization reaction and the extraction occurred simultaneously on the surface of the bar (length 7.5 mm) under acidic conditions. The method optimization was carried out by univariate and multivariate analysis. The optimal conditions for the method were a DNPH to aldehydes ratio of 40:1, buffer solution of pH 4.0, extraction time of 60 min and liquid desorption of 10 min in 100 µL of acetonitrile. The aldehydes were analyzed by HPLC-DAD with a simple and fast (6 min) chromatographic run. The limits of detection (LODs) for hexanal and heptanal were 1.00 and 0.73 µmol L-1, respectively. The relative recoveries in urine samples ranged from 88 to 111% with relative standard deviations (RSDs) being less than 7%. The method developed is of low cost and can be successfully used for the quantification of these two lung cancer biomarkers in human urine samples, potentially providing an early diagnosis of lung cancer.

Wharton's jelly cells from sheep umbilical cord maintained with different culture media are permissive to in vitro infection by Small Ruminant Lentiviruses / Células da geleia de Wharton do cordão umbilical ovino mantidas em diferentes meios de cultivo são permissivas à infecção in vitro por lentivírus de pequenos ruminantes

This study aimed to isolate cells from the Wharton's jelly of umbilical cord (WJUC) of sheep collected during natural parturition using different culture media, in addition to reporting for the first time the permissiveness of these cells to in vitro infection by small ruminant lentiviruses. Ten umbilical cords were collected from healthy sheep. Each cord explants were grown in different media consisting of MEM, low glucose DMEM, M199, and RPMI-1640. The permissiveness of infection of sheep cells from WJUC was tested with CAEV-Cork and MVV-K1514 strains, inoculating 0.1 MOI of each viral strain. Four supernatants from each strain were obtained from WJUC sheep cell cultures infected in different media. The results demonstrated the presence of cytopathic effect after the in vitro infection by CAEV-Cork and MVV-K1514 with all of the tested culture media. Nested-PCR detected proviral DNA in all supernatants. Supernatants containing CAEV-Cork viruses had TCID 50/ml titres of 10 5.5 in MEM, 10 4.0 in low glucose DMEM, 105.0 in M199, and 10 5.7 in RPMI-1640. Supernatants containing the MVV-K1514 virus had TCID 50/ml titres of 10 4.3 in MEM, 10 3.5 in low-glucose DMEM, 10 4.7 in M199, and 10 3.5 in RPMI-1640. Sheep cells from WJUC are permissive to in vitro infection by small ruminant lentivirus.(AU)

O objetivo deste estudo foi isolar células da geleia de Wharton do cordão umbilical (GWCU) ovino coletado por ocasião do parto natural, utilizando-se diferentes meios de cultivo, além de relatar, pela primeira vez, sua permissividade à infecção in vitro por lentivírus de pequenos ruminantes (LVPRs). Dez cordões umbilicais foram coletados de ovelhas hígidas e soronegativas para LVPRs pelo teste de imunodifusão em gel de agarose (IDGA). De cada cordão, explantes foram cultivados em quatro meios distintos que consistiram em MEM, DMEM baixa glicose, meio 199 e RPMI-1640, todos acrescidos de 10% de soro fetal bovino em estufa com atmosfera úmida e 5% de CO2 a 37ºC. A permissividade de infecção das células GWCU ovino foi testada frente às cepas CAEV-Cork e MVV-K1514, inoculando-se 0,1 MOI de cada cepa viral e corando as monocamadas com May Grunwald Giemsa para visualização do efeito citopático. Foram obtidos quatro sobrenadantes CAEV-Cork e quatro MVV-K1514, provenientes do cultivo de células GWCU ovino infectadas por 21 dias em meios distintos, dos quais foram realizadas titulação em membrana sinovial caprina e extração do DNA pró-viral para realização de nested-PCR e eletroforese em gel de agarose a 2%. Os resultados demonstraram a presença de efeito citopático na infecção in vitro tanto por CAEV-Cork como por MVV-K1514 em todos os meios de cultivo, sendo visualizados sincícios e lise celular em microscópio invertido. A nested-PCR detectou o DNA pró-viral tanto do CAEV-Cork como do MVV-K1514 em todos os sobrenadantes. Os sobrenadantes contendo o vírus CAEV-Cork apresentaram títulos em TCID50/mL de 10 5,5 em MEM, 10 4,0 em DMEM baixa glicose, 10 5,0 em meio 199 e 10 5,7 em RPMI-1640. Os sobrenadantes contendo o vírus MVV-K1514 apresentaram título em TCID 50/mL de 10 4,3 em MEM, 10 3,5 em DMEM baixa glicose, 10 4,7 em meio 199 e 10 3,5 em RPMI-1640. Células GWCU ovino são permissivas à infecção in vitro pelos lentivírus de pequenos ruminantes CAEV-Cork e MVV-K1514.(AU)

Wharton's jelly cells from sheep umbilical cord maintained with different culture media are permissive to in vitro infection by Small Ruminant Lentiviruses / Células da geleia de Wharton do cordão umbilical ovino mantidas em diferentes meios de cultivo são permissivas à infecção in vitro por lentivírus de pequenos ruminantes

This study aimed to isolate cells from the Wharton's jelly of umbilical cord (WJUC) of sheep collected during natural parturition using different culture media, in addition to reporting for the first time the permissiveness of these cells to in vitro infection by small ruminant lentiviruses. Ten umbilical cords were collected from healthy sheep. Each cord explants were grown in different media consisting of MEM, low glucose DMEM, M199, and RPMI-1640. The permissiveness of infection of sheep cells from WJUC was tested with CAEV-Cork and MVV-K1514 strains, inoculating 0.1 MOI of each viral strain. Four supernatants from each strain were obtained from WJUC sheep cell cultures infected in different media. The results demonstrated the presence of cytopathic effect after the in vitro infection by CAEV-Cork and MVV-K1514 with all of the tested culture media. Nested-PCR detected proviral DNA in all supernatants. Supernatants containing CAEV-Cork viruses had TCID 50/ml titres of 10 5.5 in MEM, 10 4.0 in low glucose DMEM, 105.0 in M199, and 10 5.7 in RPMI-1640. Supernatants containing the MVV-K1514 virus had TCID 50/ml titres of 10 4.3 in MEM, 10 3.5 in low-glucose DMEM, 10 4.7 in M199, and 10 3.5 in RPMI-1640. Sheep cells from WJUC are permissive to in vitro infection by small ruminant lentivirus.(AU)

O objetivo deste estudo foi isolar células da geleia de Wharton do cordão umbilical (GWCU) ovino coletado por ocasião do parto natural, utilizando-se diferentes meios de cultivo, além de relatar, pela primeira vez, sua permissividade à infecção in vitro por lentivírus de pequenos ruminantes (LVPRs). Dez cordões umbilicais foram coletados de ovelhas hígidas e soronegativas para LVPRs pelo teste de imunodifusão em gel de agarose (IDGA). De cada cordão, explantes foram cultivados em quatro meios distintos que consistiram em MEM, DMEM baixa glicose, meio 199 e RPMI-1640, todos acrescidos de 10% de soro fetal bovino em estufa com atmosfera úmida e 5% de CO2 a 37ºC. A permissividade de infecção das células GWCU ovino foi testada frente às cepas CAEV-Cork e MVV-K1514, inoculando-se 0,1 MOI de cada cepa viral e corando as monocamadas com May Grunwald Giemsa para visualização do efeito citopático. Foram obtidos quatro sobrenadantes CAEV-Cork e quatro MVV-K1514, provenientes do cultivo de células GWCU ovino infectadas por 21 dias em meios distintos, dos quais foram realizadas titulação em membrana sinovial caprina e extração do DNA pró-viral para realização de nested-PCR e eletroforese em gel de agarose a 2%. Os resultados demonstraram a presença de efeito citopático na infecção in vitro tanto por CAEV-Cork como por MVV-K1514 em todos os meios de cultivo, sendo visualizados sincícios e lise celular em microscópio invertido. A nested-PCR detectou o DNA pró-viral tanto do CAEV-Cork como do MVV-K1514 em todos os sobrenadantes. Os sobrenadantes contendo o vírus CAEV-Cork apresentaram títulos em TCID50/mL de 10 5,5 em MEM, 10 4,0 em DMEM baixa glicose, 10 5,0 em meio 199 e 10 5,7 em RPMI-1640. Os sobrenadantes contendo o vírus MVV-K1514 apresentaram título em TCID 50/mL de 10 4,3 em MEM, 10 3,5 em DMEM baixa glicose, 10 4,7 em meio 199 e 10 3,5 em RPMI-1640. Células GWCU ovino são permissivas à infecção in vitro pelos lentivírus de pequenos ruminantes CAEV-Cork e MVV-K1514.(AU)

A novel approach to bar adsorptive microextraction: Cork as extractor phase for determination of benzophenone, triclocarban and parabens in aqueous samples.

This study describes the use of cork as a new coating for bar adsorptive microextraction (BAµE) and its application in determining benzophenone, triclocarban and parabens in aqueous samples by HPLC-DAD. In this study bars with 7.5 and 15 mm of length were used. The extraction and liquid desorption steps for BAµE were optimized employing multivariate and univariate procedures. The desorption time and solvent used for liquid desorption were optimized by univariate and multivariate studies, respectively. For the extraction step the sample pH was optimized by univariate experiments while the parameters extraction time and ionic strength were evaluated using the Doehlert design. The optimum extraction conditions were sample pH 5.5, NaCl concentration 25% and extraction time 90 min. Liquid desorption was carried out for 30 min with 250 µL (bar length of 15 mm) or 100 µL (bar length of 7.5 mm) of ACN:MeOH (50:50, v/v). The quantification limits varied between 1.6 and 20 µg L(-1) (bar length of 15 mm) and 0.64 and 8 µg L(-1) (bar length of 7.5 mm). The linear correlation coefficients were higher than 0.98 for both bars. The method with 7.5 mm bar length showed recovery values between 65 and 123%. The bar-to-bar reproducibility and the repeatability were lower than 13% (n = 2) and 14% (n = 3), respectively.

Use of green coating (cork) in solid-phase microextraction for the determination of organochlorine pesticides in water by gas chromatography-electron capture detection.

A novel method for the determination of organochlorine pesticides in water samples with extraction using cork fiber and analysis by gas chromatography with electron capture detector was developed. Also, the procedure to extract these pesticides with DVB/Car/PDMS fiber was optimized. The optimization of the variables involved in the extraction of organochlorine pesticides using the aforementioned fibers was carried out by multivariate design. The optimum extraction conditions were sample temperature 75 °C, extraction time 60 min and sodium chloride concentration 10% for the cork fiber and sample temperature 50 °C and extraction time 60 min (without salt) for the DVB/Car/PDMS fiber. The quantification limits for the two fibers varied between 1.0 and 10.0 ng L(-1). The linear correlation coefficients were >0.98 for both fibers. The method applied with the use of the cork fiber provided recovery values between 60.3 and 112.7 and RSD≤25.5 (n=3). The extraction efficiency values for the cork and DVB/Car/PDMS fibers were similar. The results show that cork is a promising alternative as a coating for SPME.