Multiple names for multi-scope analytical methods, more than a semantic issue.

The use of multiple names for multi-scope analytical methods is often confusing. More precise classification to better accommodate the scope of analytical validation and application is needed.

Tannat grape pomace as an ingredient for potential functional biscuits: bioactive compound identification, in vitro bioactivity, food safety, and sensory evaluation.

Grape pomace, the main by-product of wine process, shows high potential for the development of functional foods, being a natural source of bioactive compounds and dietary fiber. Thus, the present study proposes the development of five potential functional biscuits. The five formulations were achieved by varying the Tannat grape pomace powder (TGP, 10-20% w/w total wet dough) and sweetener sucralose (2-4% w/w total wet dough) content through a factorial design with central points. TGP microbiological and pesticides analysis were performed as a food safety requirement. Identification of bioactive compounds by HPLC-DAD-MS, in vitro bioactivity (total phenol content, antioxidant by ABTS and ORAC-FL, antidiabetic and antiobesity by inhibition of α-glucosidase and pancreatic lipase, respectively) and sensory properties of the biscuits were evaluated. TGP microbiological and pesticides showed values within food safety criteria. Sensory profiles of TGP biscuits were obtained, showing biscuits with 20% TGP good sensory quality (7.3, scale 1-9) in a cluster of 37 out of 101 consumers. TGP addition in biscuits had a significant (p < 0.05) effect on total phenolic content (0.893-1.858 mg GAE/g biscuit) and bioactive properties when compared to controls: 11.467-50.491 and 4.342-50.912 µmol TE/g biscuit for ABTS and ORAC-FL, respectively; inhibition of α-glucosidase and pancreatic lipase, IC50 35.572-64.268 and 7.197-47.135 mg/mL, respectively. HPLC-DAD-MS results showed all the identified phenolic compounds in 20/4% biscuit (TGP/sucralose%) were degraded during baking. Malvidin-3-O-(6'-p-coumaroyl) glucoside, (+)-catechin, malvidin-3-O-glucoside, and (-)-epicatechin were the main phenolic compounds (in descendent order of content) found. The bioactive properties could be attributed to the remaining phenolic compounds in the biscuits. In conclusion, TGP biscuits seemed to be a promising functional food with potential for ameliorating oxidative stress, glucose and fatty acids levels with good sensory quality.

Multiresidue analysis of basic, neutral, and acidic pesticides in biobeds' biomixture.

Contain between 1 and 3 bullet points highlighting the customization rather than the steps of the procedure. An analytical methodology was adjusted and validated for the analysis of 15 selected pesticides currently employed in extensive agriculture. The main application of this methodology is studying the pesticides degradation behavior using biobeds as a friendly environmental tool for the treatment of wastewaters generated in fields. The scope of the method was selected based on the most used pesticides in soybean crops in Uruguay. The novelty of this work is the inclusion of neutral and acidic herbicides such as 2,4-D, clethodim, dicamba, together with fungicides and insecticides which are usually included in Multi Residue Methods. An acetonitrile extraction methodology without a clean-up step yielded acceptable results for all the analytes. The instrumental analysis was performed using HPLC-MS/MS. The selected methodology was validated according to the SANTE guidelines. The recoveries were between 65 and 130% with RSD < 20%. The instrumental LOQs were fixed at 1 µg/L for all the compounds except for clethodim, and the method LOQs were 1 mg/kg in biomixture dry basis. These LOQs values are acceptable for biodegradation studies in biobeds. A multiresidue methodology was: • Validated for 15 pesticides in biomixture. • Acidic herbicides were included in the scope. • The method was employed for the environmental monitoring of pesticide degradation in biobeds.

Determination of multiclass, semi-polar pesticide residues in fatty fish muscle tissue by gas and liquid chromatography mass spectrometry.

With the aim of monitoring multiclass semi-polar pesticide residues in freshwater fatty fish, two QuEChERS approaches (so-called acetate buffered and unbuffered versions) were evaluated for the determination of 77 pesticide residues. Compounds were selected according to the dominant rainfed agriculture activities in South America. Unbuffered QuEChERS was finally chosen for validation purposes owing that it provided the best results in terms of recovery yields. Method performance was evaluated in two instrumental systems, liquid chromatography - tandem mass spectrometry (LC-MS/MS) in Scheduled MRM™ algorithm available on hybrid quadrupole - linear ion trap (QLIT) instrument, and gas chromatography - mass spectrometry (GC-MS) under selected ion monitoring (SIM) mode. Spiking experiments were carried out to determine the trueness, precision, linearity, limit of quantification of the method as well as matrix effect. The Unbuffered QuECHERS method described here: •Was validated for the analysis of 67 pesticide residues in fish muscle tissue.•Presented quantification limits in the range 1-15 µg kg-1 for the vast majority of the studied compounds.•Enable environmental monitoring of pesticide residues in fish due to their low LOQs.

Phenolic Profiling and Antioxidant Capacity of Eugenia uniflora L. (Pitanga) Samples Collected in Different Uruguayan Locations.

The use of nutrient-rich foods to enhance the wellness, health and lifestyle habits of consumers is globally encouraged. Native fruits are of great interest as they are grown and consumed locally and take part of the ethnobotanic knowledge of the population. Pitanga is an example of a native fruit from Uruguay, consumed as a jelly or an alcoholic beverage. Pitanga has a red-violet pigmentation, which is a common trait for foods that are a good source of antioxidants. Hence, fruits from different Uruguayan regions were analyzed via miniaturized sample preparation method, HPLC-DAD-ESI/MSn and RP-HPLC-DAD techniques to identify and quantify phenolic compounds, respectively. The antioxidant capacity was evaluated via DPPH and ORAC (Oxygen Radical Absorbance Capacity) assays. A multivariate linear regression was applied to correlate the observed antioxidant capacity with the phenolic content. Furthermore, Principal Components Analysis was performed to highlight characteristics between the various samples studied. The main results indicated differences between northern and southern Uruguayan samples. Delphinidin-3-hexoside was present in southern samples (mean of 293.16 µmol/100 g dry weight (DW)) and absent in the sample collected in the north (sample 3). All the samples contain high levels of cyanidin-3-hexoside, but a noticeable difference was found between the northern sample (150.45 µmol/100 g DW) and the southern sample (1121.98 µmol/100 g DW). The antioxidant capacity (mean ORAC of 56370 µmol Trolox®/100 g DW) were high in all the samples compared to the Food and Drug Administration (FDA) database of similar berry-fruits. The results of this study highlight the nutraceutical value of a native fruit that has not been exploited until now.

Miniaturized QuEChERS based methodology for multiresidue determination of pesticides in odonate nymphs as ecosystem biomonitors.

The impacts of the modern, agrochemicals based agriculture that threatens the overall systems sustainability, need to be monitored and evaluated. Seeking for agroecosystems monitors, the present article focus in the occurrence and abundance of aquatic macroinvertebrates, that have been frequently used as bioindicators of water quality due to their relationship with land use. Some of these organisms are on the top of the food chain, where bioaccumulation and biomagnification processes can be observed, and they can turn into secondary pollution sources of systems and terrestrial organisms as well. Odonate nymphs, which belong to the functional group of predators, were selected for this study. A methodology to determine 73 pesticide residues in odonate nymphs by LC-MS/MS and GC-MS/MS was developed. A QuEChERS sample preparation strategy was adapted. As it is complex to obtain samples especially in disturbed ecosystems, the method was minimized to a sample size of 200mg of fresh nymphs. The method was validated and good recoveries (71-120%) with RSDs below 20% for the majority of the studied pesticides at least at two of the assayed levels 1, 10 and 50µgkg-1 were obtained. For 32 analytes the limit of quantitation was 1µgkg-1 and 10µgkg-1 for the others. The lineal range was observed between 1-100µgkg-1 in matrix-matched and solvent calibration curves for most of the assessed pesticides. LC-MS/MS matrix effects were evaluated, 40% of the analytes presented low or no signal suppression. Only flufenoxuron presented high matrix effects. The obtained methodology is adequate for pesticide multiresidue analysis in aquatic macroinvertebrates (odonates) aiming to contribute to the ecological state evaluation of freshwater ecosystems.

Determination of pesticide residues in globe artichoke leaves and fruits by GC-MS and LC-MS/MS using the same QuEChERS procedure.

Aiming to select the most suitable sample preparation for the multiresidue analysis of pesticides in globe artichoke (Cynara cardunculus L.), a modified QuEChERS, a matrix solid phase dispersion and a dispersive ethyl acetate extraction were compared. Trueness and precision were determined at 0.2mgkg-1 for the three methods. The modified QuEChERS showed the best performance. The scope of the method was enlarged to 35 GC and 63 LC amenable pesticides, its overall performance was evaluated and validated to artichoke leaves and fruits according to DG-SANTE Guidelines. Different matrix effects were observed for most of the pesticides which were higher for leaves than fruits. Difenoconazole and flutriafol suffered signal suppression in leaves extracts but showed positive matrix effects in fruits. All pesticides were analyzed at or below their Maximum Residue Levels fixed for globe artichoke by the European Union. The method was successfully applied for the analysis of commercial samples.

Antimutagenic and antioxidant properties of the aqueous extracts of organic and conventional grapevine Vitis labrusca cv. Isabella leaves in V79 cells.

Grapes are one of the most commonly consumed fruit, in both fresh and processed forms; however, a significant amount is disposed of in the environment. Searching for a use of this waste, the antigenotoxic, antimutagenic, and antioxidant activities of aqueous extracts from organic and conventional Vitis labrusca leaves were determined using V79 cells as model. The antigenotoxic activity was analyzed by the alkaline comet assay using endonuclease III and formamidopyrimidine DNA glycosylase enzymes. The antimutagenic property was assessed through the micronucleus (MN) formation, and antioxidant activities were assessed using 2',7'-dichlorodihydrofluorescin diacetate (DCFH-DA) assay and 2,2-diphenyl-1-picrylhydrazyl (DPPH(●)) radical scavenging, as well as with superoxide dismutase (SOD) and catalase (CAT) activity assays. In addition, phenolic content and ascorbic acid levels of both extracts were determined. Data showed that both organic and conventional grapevine leaves extracts possessed antigenotoxic and antimutagenic properties. The extract of organic leaves significantly reduced intracellular reactive oxygen species (ROS) levels in V79 cells, and displayed greater ability for DPPH(●) scavenging and higher SOD and CAT activities than extract from conventional leaves. Further, the extract from organic leaves contained higher phenolic and ascorbic acid concentrations. In summary, extracts from organic and conventional grape leaves induced important in vitro biological effects.

Development of analytical methodologies to assess recalcitrant pesticide bioremediation in biobeds at laboratory scale.

To assess recalcitrant pesticide bioremediation it is necessary to gradually increase the complexity of the biological system used in order to design an effective biobed assembly. Each step towards this effective biobed design needs a suitable, validated analytical methodology that allows a correct evaluation of the dissipation and bioconvertion. Low recovery yielding methods could give a false idea of a successful biodegradation process. To address this situation, different methods were developed and validated for the simultaneous determination of endosulfan, its main three metabolites, and chlorpyrifos in increasingly complex matrices where the bioconvertor basidiomycete Abortiporus biennis could grow. The matrices were culture media, bran, and finally a laboratory biomix composed of bran, peat and soil. The methodology for the analysis of the first evaluated matrix has already been reported. The methodologies developed for the other two systems are presented in this work. The targeted analytes were extracted from fungi growing over bran in semisolid media YNB (Yeast Nitrogen Based) with acetonitrile using shaker assisted extraction, The salting-out step was performed with MgSO4 and NaCl, and the extracts analyzed by GC-ECD. The best methodology was fully validated for all the evaluated analytes at 1 and 25mgkg(-1) yielding recoveries between 72% and 109% and RSDs <11% in all cases. The application of this methodology proved that A. biennis is able to dissipate 94% of endosulfan and 87% of chlorpyrifos after 90 days. Having assessed that A. biennis growing over bran can metabolize the studied pesticides, the next step faced was the development and validation of an analytical procedure to evaluate the analytes in a laboratory scale biobed composed of 50% of bran, 25% of peat and 25% of soil together with fungal micelium. From the different procedures assayed, only ultrasound assisted extraction with ethyl acetate allowed recoveries between 80% and 110% with RSDs <18%. Linearity, recovery, precision, matrix effect and LODs/LOQs of each method were studied for all the analytes: endosulfan isomers (α & ß) and its metabolites (endosulfan sulfate, ether and diol) as well as for chlorpyrifos. In the first laboratory evaluation of these biobeds endosulfan was bioconverted up to 87% and chlorpyrifos more than 79% after 27 days.

Vasorelaxant Effect of a Baccharis trimera Infusion on Precontracted Rat Aortic Rings.

Baccharis trimera (Less.) DC is a South American plant that in folk medicine is considered to produce reduction in blood pressure. One aspect of this putative effect is the vasorelaxation. The aim of this work was to evaluate the ability of a B. trimera extract to relax rat aortic rings precontracted with noradrenaline. As the infusion is the usual way of intake of this plant, an infusion of B. trimera was prepared using 100 g of the plant (leaves) boiled in water, frozen and lyophilized. Working solutions were prepared using different concentrations of the dried extract diluted in Krebs Henseleit solution. It was proved that the infusion relaxed the aortic rings in a dose dependent manner 100 minutes after adding the exract to the bath. Considering as 100% the maximum contraction achieved with noradrenaline, a relaxation of 101.1 ± 2.3% was observed with the highest dose of the infusion used in these experiments (0.32 mg/mL). While in control rings relaxation was 12.9 ± 2.4%. In aortic rings denuded from endothelium the percentage of vasoralaxation did not show statistically significant differences when compared to intact rings. These data support the hypothesis of a vasorelaxant effect of this plant and constitutes the first approach to the scientific basis of a potential antihypertensive effect.

Comparison and evaluation of two methods for the pesticide residue analysis of organophosphates in yerba mate

Abstract Microwave Assisted Extraction and a modified CEN-QuEChERS methodology were evaluated as extraction and clean up procedures for the simultaneous analysis of 42 organophosphate pesticides in yerba mate (Ilex paraguaiensis). The obtained extracts were analyzed by gas chromatography using a flame photometric detector. Linearity, recovery percentages, relative standard deviations, detection and quantification limits and matrix effects were determined according to DG-SANCO guidelines for both methods. At 0.2 and 0.5 mg/kg the evaluated methods showed percentages recoveries between 70 and 120% for most of the analytes. Using Microwave Assisted Extraction methodology, 33 pesticide residues could be properly analyzed whereas only 27 could be determined with the proposed modified QuEChERS. All relative standard deviation were below 18% except for omethoate and disulfoton sulfone when evaluated by the modified QuEChERS. The limits of detection in both methodologies were 0.2 mg/kg for most of the analyzed compounds. The average detection limit for QuEChERS was 0.04 mg/kg. For 19 of the analytes determined through Microwave Assisted Extraction the lowest validated level were 0.004 mg/kg. Signal suppression/enhancement was observed for most of the pesticides, thus matrix-matched calibration curves were used for quantification. The Microwave Assisted Extraction and QuEChERS procedures studied could detect the organophosphate pesticides above the MRL fixed for "mate" by the European Union. They have been successfully applied for the determination of organophosphate pesticide residues in commercial samples and the positives were confirmed through GC–(ITD)-MS.

Detection of pesticides in active and depopulated beehives in Uruguay.

The influence of insecticides commonly used for agricultural purposes on beehive depopulation in Uruguay was investigated. Honeycombs, bees, honey and propolis from depopulated hives were analyzed for pesticide residues, whereas from active beehives only honey and propolis were evaluated. A total of 37 samples were analyzed, representing 14,800 beehives. In depopulated beehives only imidacloprid and fipronil were detected and in active beehives endosulfan, coumaphos, cypermethrin, ethion and chlorpyrifos were found. Coumaphos was present in the highest concentrations, around 1,000 µg/kg, in all the propolis samples from active beehives. Regarding depopulated beehives, the mean levels of imidacloprid found in honeycomb (377 µg/kg, Standard Deviation: 118) and propolis (60 µg/kg, Standard Deviation: 57) are higher than those described to produce bee disorientation and fipronil levels detected in bees (150 and 170 µg/kg) are toxic per se. The other insecticides found can affect the global fitness of the bees causing weakness and a decrease in their overall productivity. These preliminary results suggest that bees exposed to pesticides or its residues can lead them in different ways to the beehive.

Acaricidal activity of Palicourea marcgravii, a species from the Amazon forest, on cattle tick Rhipicephalus (Boophilus) microplus.

Leaves of Palicourea marcgravii were extracted successively with hexane, ethyl acetate and ethanol in order to evaluate their acaricidal activity on larvae and adult stages of the cattle tick Rhipicephalus (Boophilus) microplus. The ethyl acetate extract showed the highest bioactivity of the tested extracts, which contained 0.12% monofluoroacetic acid. On engorged female, the ethyl acetate extract showed a lethal concentration 50% - LC(50)=30.08 mg ml(-1), inhibitory concentration 50% - IC(50)=5.79 mg ml(-1) and lethal time 50% - LT(50)=4.72 days; 100% reproduction was controlled at concentrations of 50 mg ml(-1) and on larvae the ethyl acetate extract showed a LC(50)=2.46 mg ml(-1). No alkaloids were detected in any of the extracts. This is the first report on the acaricidal activity of P. marcgravii extracts against R. microplus as well as the acaricidal properties of a plant species containing monofluoroacetic acid.

Antifeedant activity of citrus waste wax and its fractions against the dry wood termite, Cryptotermes brevis.

The wood protective action of citrus wax, a waste from the citrus industry that is a mixture of citrus fruit epicuticular waxes and essential oils, was evaluated against the termite Cryptotermes brevis Walker (Isoptera: Kalotermitidae). The antifeedant index (AI) of the total wax and fractions was calculated. The total citrus wax exhibited an AI50 value of 24.69 mg/cm³, the wax after hydrodistillation showed the strongest antifeedant property (AI50 11.68 mg/cm³). Fractionation of the wax and gas chromatography-mass spectrometric analysis allowed the identification of coumarins and furancoumarins as the active compounds. These results suggest the potential use of these industrial residues as a natural approach to termite control.

Development of methods for multiresidue analysis of rice post-emergence herbicides in loam soil and their possible applications to soils of different composition.

Two simple and straightforward sample preparation methods were developed for the multiresidue analysis of post-emergence herbicides in loam soil that are commonly used in rice crop cultivation. A number of strategic soil extraction and cleanup methods were evaluated. The instrumental analysis was performed by HPLC with a diode array detector. The best compromise between the recoveries (69-98%) and good repeatability (RSD < 15%) was obtained using either methanol or methanol-ethyl acetate (70 + 30, v/v) as extraction solvents without any further cleanup. For the first time, residues of metsulfuron methyl, bensulfuron ethyl, bentazone, cyhalofop butyl, pyrazosulfuron ethyl, propanil, and clomazone were analyzed simultaneously. Quinclorac and bispyribac sodium were also assayed, but their recoveries were below 50%. Both methods had an LOD of 0.7 microg/kg and could accurately determine the residues at the 2 microg/kg level. These two methods could not be applied directly to other soil types as the recoveries strongly depended on the soil composition. The developed methodologies were successfully applied in monitoring 87 real-world soil samples, in which only propanil (6 to 12 microg/kg) and clomazone (15 to 20 microg/kg) residues could be detected.

Toxicity of Piper aduncum L. (Piperales: Piperaceae) from the Amazon forest for the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

The mortality of 14-21-day-old Rhipicephalus (Boophilus) microplus larvae, and the mortality and fertility of groups of engorged adult females exposed to different concentrations of hexane, ethyl acetate and ethanol extracts of spiked pepper (Piper aduncum) were evaluated, using a completely randomized design with five treatment groups, two control groups, and two replicates for the larvae and five replicates for the adult females. Similar methodology was used to investigate the toxicity of the essential oil hydro-distillate (94.84% dillapiole) obtained from the P. aduncum crude hexane extract. The LC(50) of the hexane extract was 9.30 mg ml(-1) for larvae and the reproduction reduction ranged from 12.48% to 54.22%, while 0.1mg/ml(-1) of the essential oil induced 100% mortality in larvae. Literature reports on natural products active against R. microplus were listed and compared with the results presented here. These results indicate that P. aduncum extracts, and particularly its essential oil, are potential alternative control agents for R. microplus.