Dispersive liquid-liquid microextraction (DLLME) for determination of tricyclic antidepressants in whole blood and plasma samples and analysis by liquid chromatography with diode array detector (LC-DAD).

Microextractions have been developed for the tricyclic antidepressants (TCAs) analysis in biological matrices, including dispersive liquid-liquid microextraction (DLLME). The proposed DLLME employed 490 µL of biological sample (whole blood or plasma), which were added 15 mg of NaCl, 10 µL of medazepam as internal standard (10 µg/mL) and 100 µL of 2 M NaOH. This mixture was homogenized by vortex (2800 rpm/10 s) and 400 µL of hexane (extractor solvent) with 600 µL of methanol (dispersing solvent) were added to the sample. After the vortex step (2800 rpm/5 s), an ultrasonic bath for 300 s was employed. Then, this content was centrifuged (10 min/10000 rpm), organic phase was collected and dried under air flow. After, 30 µL of the mobile phase was used for resuspension and 20 µL is injected into LC-DAD. This method was optimized and fully validated according to UNODC and SWGTOX guidelines, reaching limits of detection equivalent to analytical methodologies that employ mass spectrometry (MS). Also, it was applied in real cases involving suspected exposure to TCAs. So, the developed DLLME for the determination of TCAs in whole blood and plasma samples proved to be a simple, reliable, robust and reproducible method that can be used in toxicology and clinical laboratories.

Phytochemical characterisation, antioxidant capacity, and in vitro toxicity of Richardia brasiliensis gomes crude extracts.

Richardia brasiliensis is a species used in folk medicine and rich in active compounds. In this study, the extracts were submitted to UHPLC-ESI-MS/MS analysis and total polyphenols, tannins, and flavonoids assays. Besides, it was determined its antioxidant capacity, oxidative stress markers and toxicological profile. Fourteen polyphenols were found and, in the dosages, a slight change in the concentrations in each extract was observed. Regarding the antioxidant capacity, the responses were different in the methods used. There was an increase in lipid peroxidation, and NO, however total ROS remained unchanged. The cells remained more than 90% viable and the extracts did not cause damage to single strands of DNA, with the exception of the crude autumn and spring extracts at 500 µg/mL. The results found in this study suggest that extracts are potentially toxic to human leukocyte cells in high concentrations; however, more studies should be performed in different cell lines.

Enhancement of the Functional Properties of Mead Aged with Oak (Quercus) Chips at Different Toasting Levels.

Consumers increasingly prefer and seek functional beverages, which, given their characteristics, provide important bioactive compounds that help prevent and treat chronic diseases. Mead is a traditional fermented alcoholic beverage made from honey solution. The aging process of mead with oak chips is innovative and bestows functional characteristics to this beverage. Thus, in this study, we sought to develop and characterize a novel functional beverage by combining the health benefits of honey with the traditional aging process of alcoholic beverages in wood. Phenolic compounds, flavonoids, and antioxidant capacity were analyzed in mead using oak chips at different toasting levels and aged for 360 days. LC-ESI-QTOF-MS/MS was used to analyze the chemical profile of different meads. Over time, the aging process with oak chips showed a higher total phenolic and flavonoid content and antioxidant capacity. Eighteen compounds belonging to the classes of organic acids, phenolic acids, flavonoids, and tannins were identified in meads after 360 days. Our findings revealed that the addition of oak chips during aging contributed to p-coumaric, ellagic, abscisic, and chlorogenic acids, and naringenin, vanillin, and tiliroside significantly impacted the functional quality of mead.

Integrative Techniques Confirms the Presence of Bemisia tabaci Parasitoids: Encarsia formosa, Encarsia porteri and Eretmocerus mundus (Hymenoptera: Aphelinidae) on Soybean and Tomatoes in South Brazil.

Parasitoid wasps from the Aphelinidae family (Hymenoptera) are important control agents of Bemisia tabaci (Gennadius, 1889) cryptic species, both through reproduction and feeding processes. Identifying native parasitoid species within agricultural systems affected by Bemisia whitefly species is the first step to developing guidelines for the creation and release of biological control agents aiming at this highly damaging pest species complex. Taxonomic and phylogenetic analyses based on morphological and molecular characters, respectively, confirmed the occurrence of Encarsia formosa (Gahan, 1924) in greenhouse tomatoes from Santa Maria, Encarsia porteri (Mercet, 1928) in open-field soybean from Santa Maria, and Eretmocerus mundus Mercet, 1931 in greenhouse tomatoes from São José do Hortêncio, all within Rio Grande do Sul state (South Brazil). This is the first report of En. formosa, En. porteri and Er. mundus parasitising B. tabaci in South Brazil, and the first En. porteri partial mtCOI gene sequence being reported and characterised. The high temperature inside the tomato greenhouses can be a possible cause for the predominance of Er. mundus in São José do Hortêncio, and sex ratios in the surveyed populations point to female and male prevalence within Encarsia and Eretmocerus genera, respectively. The combined use of taxonomic and molecular characterisation highlights the importance of combining both morphological and molecular approaches in the assessment of previously unidentified whitefly parasitoids.

Chemical study, antioxidant activity, and genotoxicity and cytotoxicity evaluation of Ruellia angustiflora.

Ruellia angustiflora (Acanthaceae) is known as flower-of-fire, and its leaves are traditionally employed to promote wound healing. This study was aimed at extracting and characterizing the chemical constituents of the extracts of R. angustiflora obtained by ultrasound-assisted and supercritical fluid extractions, and subsequently investigate their antioxidant potential and cyto-genotoxicity. The extract obtained by ultrasound (UAE-EtOH) was characterized by ultraperformance liquid chromatography-mass spectrometer (UPLC-MS), and the extract obtained via supercritical fluid (SFE-CO2) by gas chromatograph-mass spectrometer (GC-MS). The antioxidant potential was verified by the antiradical activity against the 1,1-diphenyl-2-picrylhydrazyl radical (DPPH), and the cyto-genotoxicity evaluation to test cell viability and DNA damage was performed in peripheral blood mononuclear cells (PBMC) cultures. The SFE-CO2 extract presented some fatty acids, triterpenes, tetraterpenes, tocopherols and phytosterols. The UAE-EtOH extract contained phenolic acids and flavonoids, and showed the highest antioxidant capacity. Neither extract was genotoxic or cytotoxic at the tested concentrations.

Separation of microbial oil produced by Mortierella isabellina using polymeric membranes.

The objective of this work was to concentrate, through a membrane separation process, the fatty acids from oil/solvent mixture. The oil was obtained by ultrasound-assisted extraction from freeze-dried cells of Mortierella isabellina. The concentration of the fatty acids was investigated using flat-sheet polymer membranes of ultrafiltration and nanofiltration. The effects of temperature and pressure were evaluated by the retention of the fatty acids. Oil retentions between 45.23 and 58.20% to ultrafiltration membrane and 43.50 and 56.00% to nanofiltration membrane were observed. The best condition for the ultrafiltration membrane was 4 bar and 40 °C and for nanofiltration membrane was 12 bar and 50 °C. The oil contains a high concentration of oleic acid and palmitic acid that is a desirable property for the biodiesel production. The results showed the applicability of this technology in the solvent recovery step whereas the oil recovered contains a high concentration of fatty acids.

Optimization of subcritical water hydrolysis of pecan wastes biomasses in a semi-continuous mode.

Pecan cultivation has increased in recent years. Consequently, the amount of lignocellulosic residuals from its production has expanded. Thus, there is a necessity to explore and add value to their coproducts. The objective of this work was to obtain reducing sugars from pecan biomasses by the optimization of the subcritical water hydrolysis technology in a semi-continuous mode and the physicochemical and morphological characterization of these materials, such as SEM, TGA and FT-IR analysis. Temperatures of 180, 220 and 260 °C, water/solids mass ratio of 15 and 30 g water/g biomass and total reaction time of 15 min were used. The highest reducing sugar yield was 27.1 g/100 g of biomass, obtained at 220 °C and R-15 for pecan shells. TGA, SEM and FT-IR analysis indicated the modifications of structures and compositions of biomasses in fresh and hydrolyzed samples.

One maternal lineage leads the expansion of Thaumastocoris peregrinus (Hemiptera: Thaumastocoridae) in the New and Old Worlds.

The bronze bug, Thaumastocoris peregrinus, an Australian native insect, has become a nearly worldwide invasive pest in the last 16 years and has been causing significant damage to eucalypts (Myrtaceae), including Eucalyptus spp. and Corymbia spp. Its rapid expansion leads to new questions about pathways and routes that T. peregrinus used to invade other continents and countries. We used mtDNA to characterize specimens of T. peregrinus collected from 10 countries where this species has become established, including six recently invaded countries: Chile, Israel, Mexico, Paraguay, Portugal, and the United States of America. We then combined our mtDNA data with previous data available from South Africa, Australia, and Europe to construct a world mtDNA network of haplotypes. Haplotype A was the most common present in all specimens of sites sampled in the New World, Europe, and Israel, however from Australia second more frequently. Haplotype D was the most common one from native populations in Australia. Haplotype A differs from the two major haplotypes found in South Africa (D and G), confirming that at least two independent invasions occurred, one from Australia to South Africa, and the other one from Australia to South America (A). In conclusion, Haplotype A has an invasion success over many countries in the World. Additionally, analyzing data from our work and previous reports, it is possible to suggest some invasive routes of T. peregrinus to predict such events and support preventive control measures.

Antioxidant compounds extracted from Diaporthe schini using supercritical CO2 plus cosolvent.

Endophytic fungi have been highlight in the production of secondary metabolites with different bioactive properties, such as in the production of the antioxidant compounds. Therefore, the objective of this work was the extraction of the antioxidant compounds from the biomass of Diaporthe schini using supercritical carbon dioxide (CO2) without and with ethanol as cosolvent. The biomass was produced by submerged fermentation and the parameters evaluated in the extraction process were: pressure (150-250 bar), temperature (40-60 ºC) and cosolvent [biomass: cosolvent ratio, 1:0, 1:0.75 and 1:1.5 (w/v)]. Extraction yield, antioxidant activity and chemical composition of the extracts were determined. The highest extraction yield (3.24 wt.%) and the best antioxidant activity against the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical (96.62%) were obtained at 40 ºC, 250 bar and biomass:cosolvent ratio of 1:1.5 (w/v). The chemical compounds 1,4-diaza-2,5-dioxo-3-isobutyl bicyclo[4.3.0]nonane and benzeneethanol identified in GC/MS could be responsible for the antioxidant activity found in this study.

Development of nanoemulsions containing Physalis peruviana calyx extract: A study on stability and antioxidant capacity.

The aim of this study was to develop and evaluate the physicochemical and antioxidant stability of nanoemulsions containing a Physalis peruviana calyx extract (CPp-NE) and free extracts under different storage conditions (7 and 25 °C) and with absence or incidence of light for 120 days. The calyx extracts were prepared with ethanol 60% and characterized for later preparation of the nanoemulsions by spontaneous emulsification. The formulations presented nanometric sizes, low polydispersity index, negative zeta potential, acid pH, rutin content (11 µg·mL-1), and encapsulation efficiency of 85%. Regarding the stability, the droplet size and PdI of the CPp-NE stored at refrigeration temperature in the dark, room temperature in the dark, and refrigeration temperature with light incidence were stable for 120 days and with no visible changes in the formulations. The antioxidant capacity was related to the reducing capacity, and the best results were found for nanoemulsions stored at room temperature and in absence of light. In addition, CPp-NE presented higher antioxidant and reducing capacity in relation to the free extracts.

Improving the soluble lipase-catalyzed biodiesel production through a two-step hydroesterification reaction system.

The application of lipases in liquid formulation instead of immobilized forms in the enzymatic biodiesel synthesis can make the process cost-efficient, more competitive, and sustainable. However, despite the benefits, the long reaction times required to achieve satisfactory yields is still a drawback of this biotechnological process. In this sense, employing the novel low-cost soluble NS40116 lipase, this paper proposes an innovative two-step hydroesterification reaction (TSHR) system as a technique of improving the reaction rate of an enzymatic biodiesel production. With the employment of two central composite statistical design to optimize the parameters of each of the reactions involved, the influence of the parameters "water concentration added to the reaction," "methanol-to-oil molar ratio," and "lipase load" on the process yield, besides the acid value of the samples, was investigated. After only 8 h of reaction, the highest fatty acid methyl ester yield reached was 97.1% with an acid value of 4.62 mg KOH g-1 utilizing a total of 8 wt% water, methanol-to-oil molar ratio of 6.3:1, and 0.70 wt% of lipase. Furthermore, the statistical models for both reactions indicated to be significant with 95% of reliability. Considering that the papers published using soluble lipases in a one-step batch process normally reach similar yields to those obtained in this research after 16 h to 24 h of reaction, the proposed system demonstrated to be a promising option of process configuration for the enzymatic production of biodiesel.

Enzyme-Catalyzed Production of FAME by Hydroesterification of Soybean Oil Using the Novel Soluble Lipase NS 40116.

The performance of lipase NS 40116, a novel and promising soluble enzyme obtained from modified Thermomyces lanuginosus microorganism, was investigated in the production of biodiesel (fatty acid methyl esters-FAME) by hydroesterification. In order to investigate the potential of the biocatalyst in its soluble form, this work reports the effect of water content and enzyme load, as well as the recovery and reuse of the biocatalyst. A FAME yield of 94.30% after 12 h was achieved at 35 °C by combining 0.50 wt% of lipase, 15 wt% of water, and a methanol:oil molar ratio of 4.5:1. The analysis of the time course reaction suggests that the triacylglycerides (TAGs) are hydrolyzed by the enzyme in a first step, generating free fatty acids (FFAs), followed by the esterification of these FFAs into FAME. In relation to the reusability assays, the lipase kept approximately 90% of its catalytic activity after five cycles of reuse. In this context, the findings of this study demonstrate that lipase NS 40116 can efficiently catalyze hydroesterification reactions under mild conditions, arising as a competitive alternative for biodiesel synthesis.

Antifungal and antibacterial activity of extracts produced from Diaporthe schini.

The aim of this work was the production of bioactive metabolites by submerged fermentation from the fungus Diaporthe schini, followed by their extraction, separation and characterization. Different solvents (methanol, dichloromethane and hexane) were used for the extraction of metabolites from the fermentation broth and the extracts obtained were evaluated by in vitro antibacterial and antifungal activity. The separation and characterization of the extract from the hexane extraction was performed by column chromatography and GC-MS, respectively. The extracts had a great inhibitory action on the Gram-positive bacteria Staphylococcus epidermidis and Staphylococcus aureus, on the Gram-negative bacteria Enterobacter aerogenes and Klebsiella pneumoniae and on the fungus Candida krusei. The main metabolites produced were: 13-docosenamide, (Z)-; 2-hexadecene, 3,7,11,15-tetramethyl; 9-octadecenamide and 11-octadecenoic acid. Studies related to the antibacterial and antifungal activities of metabolites extracted from microorganisms are found in the literature. However, works about the identification of metabolites produced by submerged fermentation from Diaporthe schini were not found until the present moment. This work is an initial study where the conditions of the process can be optimized by looking for the production of a specific compound and can be a promising source for obtaining new drugs.

Obtaining fermentable sugars and bioproducts from rice husks by subcritical water hydrolysis in a semi-continuous mode.

This work aimed at producing fermentable sugars and bioproducts from rice husks by subcritical water hydrolysis at 25 MPa in a semi-continuous mode. The influences of temperature (180 °C; 220 °C; 260 °C) and liquid/solid ratio (7.5 g water/g husks; 15 g water/g husks) on reducing sugar yield (YRS), efficiency (E), kinetic profiles (0-15 min), composition of sugars, inhibitors and organic acids, and physicochemical characteristics of the remaining solid material were evaluated and discussed in the work. The highest YRS (18.0 ±â€¯2.9 g/100 g husks) and E (39.5 ±â€¯1.7 g sugars/100 g carbohydrates) were obtained at 220 °C and 7.5 gwater/g husks. In such condition, the hydrolyzed solutions presented cellobiose (18.0 g/L), xylose 17.7 g/L), arabinose (3.6 g/L), glucose (1.5 g/L), and levulinic acid (0.7 g/L). The fermentable sugars and bioproducts can be applied in several industrial fields, especially for the production of bioethanol and other higher value-added chemical compounds.

First report of the production of a potent biosurfactant with α,ß-trehalose by Fusarium fujikuroi under optimized conditions of submerged fermentation.

Biosurfactants have many advantages over synthetic surfactants but have higher production costs. Identifying microorganisms with high production capacities for these molecules and optimizing their growth conditions can reduce cost. The present work aimed to isolate and identify a fungus with high biosurfactant production capacity, optimize its growth conditions in a low cost culture medium, and characterize the chemical structure of the biosurfactant molecule. The fungal strain UFSM-BAS-01 was isolated from soil contaminated with hydrocarbons and identified as Fusarium fujikuroi. To optimize biosurfactant production, a Plackett-Burman design and a central composite rotational design were used. The variables evaluated were pH, incubation period, temperature, agitation and amount of inoculum in a liquid medium containing glucose. The partial structure of the biosurfactant molecule was identified by nuclear magnetic resonance spectrometry. F. fujikuroi reduced surface tension from 72 to 20mNm-1 under the optimized conditions of pH 5.0, 37°C and 7 days of incubation with 190rpm agitation. The partial identification of the structure of the biosurfactant demonstrated the presence of an α,ß-trehalose. The present study is the first report of the biosynthesis of this compound by F. fujikuroi, suggesting that the biosurfactant produced belongs to the class of trehalolipids.

First report of the production of a potent biosurfactant with ,-trehalose by Fusarium fujikuroi under optimized conditions of submerged fermentation

Abstract Biosurfactants have many advantages over synthetic surfactants but have higher production costs. Identifying microorganisms with high production capacities for these molecules and optimizing their growth conditions can reduce cost. The present work aimed to isolate and identify a fungus with high biosurfactant production capacity, optimize its growth conditions in a low cost culture medium, and characterize the chemical structure of the biosurfactant molecule. The fungal strain UFSM-BAS-01 was isolated from soil contaminated with hydrocarbons and identified as Fusarium fujikuroi. To optimize biosurfactant production, a PlackettBurman design and a central composite rotational design were used. The variables evaluated were pH, incubation period, temperature, agitation and amount of inoculum in a liquid medium containing glucose. The partial structure of the biosurfactant molecule was identified by nuclear magnetic resonance spectrometry. F. fujikuroi reduced surface tension from 72 to 20 mN m1 under the optimized conditions of pH 5.0, 37 °C and 7 days of incubation with 190 rpm agitation. The partial identification of the structure of the biosurfactant demonstrated the presence of an ,-trehalose. The present study is the first report of the biosynthesis of this compound by F. fujikuroi, suggesting that the biosurfactant produced belongs to the class of trehalolipids.

First report of the production of a potent biosurfactant with α, ß-trehalose by Fusarium fujikuroi under optimized conditions of submerged fermentation

Abstract Biosurfactants have many advantages over synthetic surfactants but have higher production costs. Identifying microorganisms with high production capacities for these molecules and optimizing their growth conditions can reduce cost. The present work aimed to isolate and identify a fungus with high biosurfactant production capacity, optimize its growth conditions in a low cost culture medium, and characterize the chemical structure of the biosurfactant molecule. The fungal strain UFSM-BAS-01 was isolated from soil contaminated with hydrocarbons and identified as Fusarium fujikuroi. To optimize biosurfactant production, a Plackett-Burman design and a central composite rotational design were used. The variables evaluated were pH, incubation period, temperature, agitation and amount of inoculum in a liquid medium containing glucose. The partial structure of the biosurfactant molecule was identified by nuclear magnetic resonance spectrometry. F. fujikuroi reduced surface tension from 72 to 20 mN m−1 under the optimized conditions of pH 5.0, 37 °C and 7 days of incubation with 190 rpm agitation. The partial identification of the structure of the biosurfactant demonstrated the presence of an α,β-trehalose. The present study is the first report of the biosynthesis of this compound by F. fujikuroi, suggesting that the biosurfactant produced belongs to the class of trehalolipids.