Benzo(b)fluoranthene and benzo(k)fluoranthene removal by microalgae in the presence of triazine herbicides: Matrix solid-phase dispersion and solid-phase extraction (MSPD/SPE) for HPLC-UV analysis of the different culture components.

Over the last decade, human activities in the industrial and agricultural sectors have significantly increased the concentration of persistent and harmful pollutants in aquatic ecosystems. The use of microorganisms is a green strategy for the bio-removal of certain contaminants. However, other pollutants in the same ecosystems can reduce their degrading activity and even affect their survival. Therefore, this study aimed to evaluate the efficiency of benzo(b)fluoranthene (BbF) and benzo(k)fluoranthene (BkF) removal by Selenastrum capricornutum in the presence of triazine herbicides, compounds mainly used in broadleaf weeds. The interest of this work focused on identifying in which of the microalgal components the degrading activity is best evidenced and affected. For this purpose, the use of solid-phase extraction (SPE) and matrix solid-phase dispersion (MSPD) extraction procedures and HPLC-UV analysis allowed the BbF and BkF trace quantification in biomass, liquid medium, and cell lysate separately from cultures exposed to these polycyclic aromatic hydrocarbons (PAHs) alone or with herbicides. The recovery percentages were between 78 and 94 %, good linearity (r2 ≈ 0.99), precision values measured as RSD < 15 %, and limits of detection (LOQs) at levels of ng mL-1 and ng mg-1 were obtained. The individual PAH amounts measured in the components of microalgae cultures show similar removal kinetics (removal percentages: 82-89 %). Likewise, the analysis demonstrated that the removal of PAHs is not affected in the presence of triazine herbicides (atrazine and cyanazine) and with similar removal percentages (79-86 %) compared to those cultures exposed to individual PAHs (74-83 %). These results support the possible real-world applications of PAH removal by extracts from S. capricornutum in aquatic environments contaminated with PAHs and near agriculture areas where triazine herbicides are used.

Coriander (Coriandrum sativum L.) from Alentejo (South Portugal)-Ethnobotany and Potential Industrial Use.

Coriander is a medicinal and aromatic plant (MAP) traditionally cultivated and used in Alentejo, Portugal. However, few publications are available about its ethnobotanical applications. Four independent ethnobotanical surveys were carried out: throughout the region (2002-2003), in three villages (2013), and in city markets (2007 and 2022). Coriander was the most common fresh cultivated MAP (75% of the total area) and also the most representative MAP fresh herb in city markets. The leaves, mostly, were used fresh or frozen or transformed in piso. Some of the recipes have agro-industrial potential, such as piso and aromatized olive oil. Coriander essential oils (EOs) were isolated by hydrodistillation from aerial parts with inflorescence emergence (APIs) and from fruits, and fatty acids (FA) by solvent extraction from the fruits. Gas chromatography and gas chromatography-mass spectrometry analysis showed EOs dominated by n-decanal (21-24%), 2-trans-decenal (12-18%) and n-nonane (10-17%) in APIs, and linalool dominance (73-78%) in the fruits. Petroselinic acid (32-55%) was the dominant fatty acid. A literature survey on conventional and nonconventional extraction techniques showed a constancy in the dominant compounds isolated, highlighted piso as a home-made green-extraction procedure, but also reflected the relevance of coriander as a MAP with diverse industrial potential uses.

Phytochemical compounds with promising biological activities from Ascophyllum nodosum extracts using microwave-assisted extraction.

Phytochemical-rich antioxidant extracts were obtained from Ascophyllum nodosum (AN) using microwave-assisted extraction (MAE). Critical extraction factors such as time, pressure, and ethanol concentration were optimized by response surface methodology with a circumscribed central composite design. Under the optimal MAE conditions (3 min, 10.4 bar, 46.8 % ethanol), the maximum recovery of phytochemical compounds (polyphenols and fucoxanthin) with improved antioxidant activity from AN was obtained. In addition, the optimized AN extract showed significant biological activities as it was able to scavenge reactive oxygen and nitrogen species, inhibit central nervous system-related enzymes, and exhibit cytotoxic activity against different cancer cell lines. In addition, the optimized AN extract showed antimicrobial, and anti-quorum sensing activities, indicating that this extract could offer direct and indirect protection against infection by pathogenic microorganisms. This work demonstrated that the sustainably obtained AN extract could be an emerging, non-toxic, and natural ingredient with potential to be included in different applications.

Utilization of Extraction Procedures for Evaluating Environmental Risk from Waste Materials.

Several procedures for extracting content from different waste materials types were investigated, with the aim of evaluating their environmental impact. The waste materials consisted of wastes from bauxite ore processing by means of the Bayer process (red mud, Ajka, Hungary), bauxite ore using the sintering process followed by the Bayer process (brown-red mud, Ziar nad Hronom, Banská Bystrica region, Slovakia) and sulphide ores (metal-rich post-flotation tailing, Lintich, Slovakia). The extraction procedures were carried out with the aim of isolating "mobilizable" fractions using 0.05 M ethylenediaminetetraacetic acid (EDTA) and 0.43 M acetic acid (AA) (representing environmental risk during changes in normal environmental conditions) and "maximum potentially mobilizable" fractions using 2 M HNO3 (representing the total environmental risk). The content of chosen toxic heavy metals (THMs) (Cd, Cr, Cu, Pb, Ni, Zn) and Fe, Mn as metals creating Fe/Mn oxides in the extracts and solutions after microwave digestion was determined using high-resolution continuum source flame atomic absorption spectrometry (HR CS FAAS). On the basis of the results obtained in this study, it is possible to state that different origin of waste materials is reflected in different mobility of toxic heavy metals into the surrounding environment. From the point of view of toxic heavy metals mobility, disposal site of wastes after bauxite processing are much less of a threat to the environment than disposal site of flotation sludge after processing sulphide ores. The single extraction of 0.43 M AA is more effective than the extraction of 0.05 M EDTA for the purposes of determining the content of metals in the mobilizable fraction of tailing waste materials. The mobility of the studied toxic heavy metals in the Lintich tailing decreases in the direction from the lagoon to the dam, which may indicate the fact that the dam serves to a certain extent to inhibit the mobility of metals into the surrounding ecosystem.

Cannabis Extraction Technologies: Impact of Research and Value Addition in Latin America.

The Cannabis genus of plants has been widely used in different cultures for various purposes. It is separated into three main species: sativa, indica, and ruderalis. In ancient practices, the plant was used as a multipurpose crop and valued for its fiber, food, and medicinal uses. Since methodologies for the extraction, processing, and identification of components have become available, medical, and food applications have been increasing, allowing potential development in the pharmaceutical and healthy functional food industries. Although the growing legalization and adoption of cannabis for the treatment of diseases are key factors pushing the growth of its market, the biggest challenge is to obtain higher-quality products in a time- and cost-effective fashion, making the process of extraction and separation an essential step. Latin American countries exhibit great knowledge of extraction technologies; nevertheless, it is still necessary to verify whether production costs are economically profitable. In addition, there has been an increase in commercial cannabis products that may or may not be allowed, with or without quality fact sheets, which can pose health risks. Hence, legalization is mandatory and urgent for the rest of Latin American countries. In this article, the phytochemical compounds (cannabinoids, terpenes, and phenolic compounds), the current status of legalization, extraction techniques, and research advances in cannabis in Latin America are reviewed.

Effect of incubation time of three single extraction procedures on trace element extraction from sediment and soil.

The use of standard single-extraction procedures to evaluate the mobility and availability of trace elements is a common practice in most laboratories dealing with soil or sediment analysis. Most standard single-extraction procedures describe incubations last for 2 h. However, these were tested and validated for soil analysis. Applying them for sediment analysis without further investigation might be misleading and should be reviewed with care. This paper investigates the effect of incubation time on the extraction efficiency of three standard single-extracting reagents (0.01 M CaCl2, 1 M NH4NO3, and 0.05 M EDTA). Incubation experiments with sediment and soil samples lasting for 2 h, 10 h, and 10 d were performed. The results indicated that 2 h appears sufficient to reach equilibrium using CaCl2 or NH4NO3 for soil analysis; but when analyzing sediments, incubation for 10 d resulted in higher concentrations. Incubation experiments with 0.05 M EDTA showed that incubation for 2 h was enough to extract Cd from the soil sample, Mn and to a lesser extent Cd from the sediment samples; while for the other elements, incubation for 10 d yielded higher concentrations for both sample types compared to that obtained after 2 h and 10 h separately. Relative to the pseudo-total metal contents, more than 55% of all studied elements were extracted by using 0.05 M EDTA, indicating high bioavailable metal fraction.

Insights into using green and unconventional technologies to recover natural astaxanthin from microbial biomass.

Microorganisms such as bacteria, microalgae and fungi, are natural and rich sources of several valuable bioactive antioxidant's compounds, including carotenoids. Among the carotenoids with antioxidant properties, astaxanthin can be highlighted due to its pharmaceutical, feed, food, cosmetic and biotechnological applications. The best-known producers of astaxanthin are yeast and microalgae cells that biosynthesize this pigment intracellularly, requiring efficient and sustainable downstream procedures for its recovery. Conventional multi-step procedures usually involve the consumption of large amounts of volatile organic compounds (VOCs), which are regarded as toxic and hazardous chemicals. Considering these environmental issues, this review is focused on revealing the potential of unconventional extraction procedures [viz., Supercritical Fluid Extraction (SFE), Ultrasound-Assisted Extraction (UAE), Microwave-Assisted Extraction (MAE), High-Pressure Homogenization (HPH)] combined with alternative green solvents (biosolvents, eutectic solvents and ionic liquids) for the recovery of microbial-based astaxanthin from microalgae (such as Haematococcus pluvialis) and yeast (such as Phaffia rhodozyma) cells. The principal advances in the area, process bottlenecks, solvent selection and strategies to improve the recovery of microbial astaxanthin are emphasized. The promising recovery yields using these environmentally friendly procedures in lab-scale are good indications and directions for their effective use in biotechnological processes for the production of commercial feed and food ingredients like astaxanthin.

Surface Microanalysis and Sequential Chemical Extraction as Tools for Reliable Environmental Mobility Assessment of Sb and Other Metals.

Sequential extraction procedures (SEPs) are widely used in environmental studies to infer the chemical and/or mineralogical forms of pollutants of concern in soils and sediments. Although there is no general agreement among the scientific community, these methods have shown some limitations, especially those with a lack of objectivity in their interpretation. In this work, a soil sampling campaign was carried out in an area affected by an abandoned Sb mine. Samples (0-15 cm) were carefully prepared and analyzed by an SEP. They were also studied by conventional mineralogical methods (optical and electronic microscopy, both scanning and transmission, with a coupled energy dispersive X-ray (EDX) spectrometer). When comparing the results obtained from both techniques, some discrepancies are highlighted, with As, Cu, Pb, Sb and Zn as elements of concern. For Sb, Cu, Pb and As the predominant fraction (excluding the residual one) is that associated with organic matter (from 8.54 for Sb to 18.90% for Cu). The fractions of pollutants linked to Fe and/or Mn oxides are quantitatively important for As, Pb and Zn (6.46%, 12.05% and 7.43%, respectively) and almost negligible for Cu and Sb. On the contrary, analyses carried out by EDX at a grain scale pointed out that no detectable quantities of the elements of concern were present on the surface of the organic particles. Sb and Pb were always detectable in Fe oxides (up to 1.84 and 5.76%, respectively). Regarding the role of the clayey fraction, the only disagreement between the employed SEP and the microanalysis is in relation to As. Arsenic bound to clay minerals was found to be an order of magnitude lower than As associated with Fe oxides (0.56% and 6.46%, respectively); in contrast, EDX microanalyses showed similar As contents in both groups. Given the objectiveness of EDX microanalysis, these differences should be considered inaccuracies in the interpretation of the sequential extraction results.

Cytotoxicity of an Innovative Pressurised Cyclic Solid-Liquid (PCSL) Extract from Artemisia annua.

Therapeutic treatments with Artemisia annua have a long-established tradition in various diseases due to its antibacterial, antioxidant, antiviral, anti-malaria and anti-cancer effects. However, in relation to the latter, virtually all reports focused on toxic effects of A. annua extracts were obtained mostly through conventional maceration methods. In the present study, an innovative extraction procedure from A. annua, based on pressurised cyclic solid-liquid (PCSL) extraction, resulted in the production of a new phytocomplex with enhanced anti-cancer properties. This extraction procedure generated a pressure gradient due to compressions and following decompressions, allowing to directly perform the extraction without any maceration. The toxic effects of A. annua PCSL extract were tested on different cells, including three cancer cell lines. The results of this study clearly indicate that the exposure of human, murine and canine cancer cells to serial dilutions of PCSL extract resulted in higher toxicity and stronger propensity to induce apoptosis than that detected by subjecting the same cells to Artemisia extracts obtained through canonical extraction by maceration. Collected data suggest that PCSL extract of A. annua could be a promising and economic new therapeutic tool to treat human and animal tumours.

Study on the possibility of developing food-grade hydrophobic bio-aerogels by using an oleogel template approach.

The feasibility of producing food-grade hydrophobic bio-aerogels by supercritical-carbon dioxide (SC-CO2) extraction of oil from oleogels was investigated for the first time. Medium chain triglycerides (MCT) oil was gelled using ethylcellulose (EC) at increasing concentration (10, 15, 20% w/w) and grade (EC20, 45, 100), eventually in combination with fillers. Different SC-CO2 oil extraction procedures were tested. The acquired results show that both oleogel formulation and extraction conditions can steer the EC scaffold structure. The increase in EC concentration and grade resulted in oleogels more structurally stable to SC-CO2 extraction. The application of a pulsed extraction procedure allowed obtaining a low-density (0.39 â€‹g/cm3) EC scaffold presenting 60% oil. Addition of freeze dried lettuce powder improved macrostructure homogeneity. The obtained results lay the foundations for developing food-grade hydrophobic bio-aerogels, which are expected to present unique oil absorption and bioactive delivery features.

Evaluation of stabilizing material and stabilization efficiency through comparative study of toxic heavy metal transfer between corn and peanut grown in stabilized field soil.

Soil contaminated with toxic heavy metals (THMs) was stabilized by adding a combination of waste resources in 7.0 wt%, including coal-mine drainage sludge, waste cow bone, and steelmaking slag, in the ratio of 5:35:60. Subsequently, corn and peanut were cultivated in treated soil to investigate the effects of the waste resources on THM mobility in soil and translocation to plants. Sequential extraction procedures (SEP) was used to analyze mobile phase THMs which could be accumulated in the plants. SEP shows that mobile Pb, Cd, Cu, Zn, Ni, Cr, and As were reduced by 8.48%, 29.22%, 18.85%, 21.66%, 4.58%, 62.78%, and 20.01%, respectively. The bioaccumulation of THMs was clearly hindered by stabilization; however, the increment in the amount of immobile-phase THMs and change in the amount of translocated THMs was not proportional. The corn grains grown above the soil surface were compared with the peanut grains grown beneath the soil surface, and the results indicating that the efficiency of stabilization on THM translocation may not depend on the contact of grain to soil but the nature of plant. Interestingly, the results of bioaccumulation with and without stabilization showed that the movement of some THMs inside the plants was affected by stabilization.

Selecting efficient methodologies for estimation of As and Hg availability in a brownfield.

The determination of soil metal(loid) availability presents controversy and there is no consensus or uniformity on used analytical methods. In this study nine single extraction methods (H2O, CaCl2, NaNO3, NH4NO3, DTPA, EDTA, HCl, LMWOA, TCLP) and four sequential extraction procedures (Tessier, BCR, Wenzel and Fernández-Martínez) have been compared to estimate the availability of As and Hg in two soils from a highly polluted brownfield, especially with As. The metal(loid) concentrations were also determined in three native plant species (Lotus corniculatus, Betula celtiberica and Dactylis glomerata) collected in the habitat under study. Each single extractant showed a particular capacity of As/Hg extraction because they do not extract the same forms of each element. The availability of As and Hg depended on the element characteristics, soil properties, type of extractant and degree of pollution, thus the use of a single extraction procedure provides limited information of metal(loid) availability and to reach general conclusions is difficult. Regarding the sequential extractions, each procedure showed a specific pattern for As and Hg regardless of the soil. Thus, the choice of one or other method depends on the environmental conditions, metal(loid) and soil properties. In risk assessment studies it would be recommendable to select one of the more aggressive extractants, so as not to underestimate the environmental risk. In this regard, the sequential extraction procedures render more detailed information about metal(loid) potential availability in relation to soil properties. The analysis of native plant species showed higher metal(loid) concentrations in roots than in aerial parts and differences were observed depending on the metal(loid) and the species. In general, plants showed a higher BCFs for Hg than As even though the total and available As concentrations were higher than those found for Hg, which highlights the influence of plant species on the metal(loid) uptake.

[Evaluation of different procedures for the extraction of phenolic compounds from a medicinal plant: Verbena officinalis]. / Évaluation de différents procédés d'extraction des composés phénoliques d'une plante médicinale : Verbena officinalis.

Since ancient times, plants have been the main source of bioactive molecules, such as phenolic compounds, capable of remedying various diseases. However, polyphenols' content and efficiency vary greatly as a function of several intrinsic and extrinsic factors. To optimize the procedure for the extraction of active molecules from the medicinal plant Verbena officinalis, effects of the plant origin, selected solvent, and extraction method were assessed. V. officinalis aerial parts were collected in two different regions of Tunisia (Bizerte and Ain Draham), and their bioactive molecules were extracted by maceration, decoction, and by the Soxhlet apparatus, either with water or with ethanol. Significant variability in the extracts' contents of phenolic compounds as well as their antioxidant and antimicrobial capacities were noted depending on the different studied factors. In particular, ethanol extracts were found to generally contain higher concentrations of phenolic compounds and more potent antioxidant capacities than water extracts. However, when tested against various pathogenic bacteria, water extracts were most often at least as active as ethanol extracts to inhibit bacteria growth in vitro. Finally, differences were also observed between V. officinalis samples from Bizerte compared to Ain Draham area. All of these results emphasize the need of adapting various parameters for the optimal extraction of bioactive molecules from a medicinal plant such as V. officinalis.

Title: Évaluation de différents procédés d'extraction des composés phénoliques d'une plante médicinale : Verbena officinalis. Abstract: Depuis la nuit des temps, les plantes ont été la source principale de molécules bioactives, tels les composés phénoliques, capables de remédier à diverses maladies. Cependant, le contenu et l'activité des polyphénols dépendent d'un certain nombre de facteurs intrinsèques et extrinsèques. Dans le but d'optimiser les procédés d'obtention des principes actifs de la verveine (Verbena officinalis), les effets de la provenance de la plante, du solvant et de la méthode d'extraction ont été évalués. Ainsi, la partie aérienne de V. officinalis a été collectée dans deux régions différentes de la Tunisie (Bizerte et Ain Draham). L'extraction a été réalisée par macération, décoction et par l'appareil de Soxhlet tantôt avec de l'eau tantôt avec de l'éthanol pur. Une variabilité significative des teneurs en composés phénoliques ainsi que des capacités antioxydantes et antimicrobiennes des extraits a été observée en fonction des facteurs étudiés. De manière générale, les extraits éthanoliques sont plus riches en composés phénoliques et présentent des activités antioxydantes plus fortes que les extraits aqueux. Cependant, vis-à-vis de différentes souches pathogènes, les extraits aqueux sont souvent au moins aussi puissants que les extraits éthanoliques pour inhiber la croissance bactérienne in vitro. De plus des différences notables sont observées selon que V. officinalis provient de la région de Bizerte ou d'Ain Draham. Ces résultats montrent que la prise en compte de plusieurs paramètres est nécessaire pour optimiser l'efficacité des procédures d'extraction des molécules bioactives de V. officinalis.

Effect of extraction procedures on the chemical structure, antitumor and anticoagulant properties of ulvan from Ulva lactuca of Tunisia coast.

The effect of extraction procedures on chemical composition, structural, antitumor and anticoagulant properties of the sulphated polysaccharide 'ulvan' from the green seaweed Ulva lactuca were investigated. The structural features of ulvans were carried out by FTIR and by one- and two- dimensional 1H and 13C NMR spectroscopic. The ulvans were mainly composed of rhamnose, xylose, and uronic acid. Chemical and spectroscopic analyses demonstrated that ulvans were constituted of (1→4)-ß-glucuronic acid, (1→3,4)-α-L-rhamnose-3-sulphate and (1→4)-α-xylose. The extraction procedures effect were observed in chemical structure, Mw and biological activities. Cytotoxic activity of enzymatic-chemical extract on cervical cancer cells (HeLa) (IC50 = 1000 µg/mL) was higher than on normal peripheral blood lymphocytes cells (PBL). Acid extracts promoted to reduce HeLa cells and to grow PBL cells. At high concentrations, acid extracts showed the highest APTT and TT clotting time. Antitumoral and anticoagulant activities of ulvans from Ulva lactuca promote their use as effective therapeutic agent.

Microfluidic-enabled versatile hyphenation of electromembrane extraction and thin film solid phase microextraction.

In the present study, a versatile combination of electromembrane extraction (EME) with thin film solid phase microextraction (TF-SPME) was introduced using a microfluidic chip device. The device consisted of two single channels on two separate layers. The upper channel was dedicated to donor phase flow pass, while the beneath channel was used as a reservoir for stagnant acceptor solution. A slide of fluorine doped tin oxide (FTO) was accommodated in the bottom of the acceptor phase channel. A thin layer of polyaniline was electrodeposited on the FTO surface to achieve the required thin film for TF-SPME. A stainless-steel wire was embedded in the donor phase channel and another wire was also attached to the FTO surface. The channels were separated by a piece of polypropylene membrane impregnated with 1-octanol and the whole chip was fixed with bolts and nuts. The driving force for the extraction was an 8 V direct current (DC) voltage applied across the supported liquid membrane (SLM). Under the influence of the electrical field, analytes immigrated from sample towards the acceptor phase and then adsorbed on the thin film of the solid phase. Finally, the analytes were desorbed by successive movement of a desorption solvent in the acceptor phase channel followed by injection of the desorption solution to HPLC-UV. The applicability of the proposed device was demonstrated by the determination of four synthetic food dyes: Amaranth, Ponceau 4R, Allura Red, and Carmoisine, as the model analytes. The effective parameters on the efficiency of the both EME and TF-SPME were investigated. Under the optimized conditions, the microchip provided low LODs (1-10 µg L-1), and a wide linear dynamic range of 10-1000 µg L-1 for all analytes. The system also offered RSD values lower than 5.5% and acceptable reusability of the thin film for multiple extractions.

Microplastic in marine environment: reworking and optimisation of two analytical protocols for the extraction of microplastics from sediments and oysters.

Marine sediments and sessile biota (i.e. oysters) are nowadays recognised to be affected by microplastic (MP) pollution. NOAA proposes two distinct MP extraction protocols for sandy and bed sediments, which, however, were already demonstrated to suffer from many limitations. Conversely, to what concern oysters, works already published are usually time consuming, requiring a KOH 24-48 h oxidation step. The aim of this study is to show how iterative adaptation of the NOAA protocol allows to extract MPs, included PET, from marine sediments, regardless their characteristics. The method tested on PE-LD/PET/PA/PE-HD is based on density separation and oxidation treatments which were both carefully tuned, obtaining final recoveries higher than 85% for all the micropolymers (100% for PE and PA). Furthermore, a new protocol for the extraction of MPs from oysters was assessed, highlighting its efficacy (recoveries higher than 84% for all the plastics) and time-saving peculiarity. Finally, both protocols were successfully applied in the MPs extraction from real samples from Atlantic Ocean.•The extraction of PE-LD/PET/PA/PE-HD was optimised in sediments (regardless their characteristics) and oysters.•For sediments, density separation and oxidation procedures were carefully optimised.•For oysters, oxidation times were reduced from 24 to 48 h to 1 h.

Chemical Profile, Antioxidant, Anti-Inflammatory, and Anti-Cancer Effects of Italian Salvia rosmarinus Spenn. Methanol Leaves Extracts.

In this study, we evaluated and compared the chemical composition, the antioxidant, anti-inflammatory, and anti-proliferative effects of four methanol extracts (R1-R4), of Salvia rosmarinus Spenn. in two different sites of Southern Italy obtained by maceration or ultrasound-assisted extraction. Extracts of S. rosmarinus collected on the Ionian coast are indicated with the abbreviations R1 (maceration) and R2 (ultrasound-assisted extraction). Extracts of S. rosmarinus collected on the Tyrrhenian coast are indicated with the abbreviations R3 (maceration) and R4 (ultrasound-assisted extraction). The chemical composition was analyzed using High Pressure liquid chromatography-Diod-Array detection-Electrospray ionization-Quadrupole-Mass Spectroscopy (HPLC-DAD-ESI-Q-MS). The antioxidant activity was analyzed by 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) 2,2-diphenyl-1-picrylhydrazyl (DPPH), ß-carotene bleaching, and Ferric Reducing Antioxidant Power (FRAP) assays. Antioxidant features were also assessed in lipopolysaccharide (LPS)-stimulated RAW-264.7 murine macrophages, evaluating Reactive Oxygen Species (ROS) production; in the same experimental model, the anti-inflammatory activity of the extracts was investigated. Interestingly, all extracts displayed antioxidant and anti-inflammatory properties. They exhibited significative nitrite production inhibitory activity, whith IC50 values ranging from 3.46 to 5.53 µg/mL, without impairing cell viability. The anti-inflammatory activity was also investigated by Western Blotting and immunofluorescence assay, highlighting the R3 and R4 extracts ability to reduce NF-κB translocation, as well as to disrupt the MAPKs signaling pathway. Extracts exhibited both potential anti-proliferative activity on breast cancer cells, inducing apoptosis, without affecting non-tumorigenic cells, and the ability to inhibit MDA-MB-231 cells' motility. Finally, the rosemary extracts treatment significantly reduced the power of conditioned media, from MCF-7 or MDA-MB-231 cells to induce nitrite production on RAW 264.7 cells, confirming their promising anti-inflammatory activity.

Evaluation of Removal Behavior of Cesium in Contaminated Soil Based on Speciation Analysis.

The removal efficiency of Cs from contaminated soil depends on its chemical species bound with the soil components. Therefore, in this study, we observed the elution behavior of Cs based on speciation analysis in a Cs removal experiment conducted on contaminated soils. The treatment method was optimized using simulated contaminated soil and applied to actual contaminated soil on a large scale as well. The elution rate of Cs was approximately 50% or more in both actual and simulated contaminated soil using the optimized treatment method. From the obtained results, a robust treatment method using an eluting reagent and a magnetic adsorbent with low energy costs is proposed. Additionally, the usefulness of speciation analysis in decontamination studies was confirmed.

Contribution of Flavonoids and Iridoids to the Hypoglycaemic, Antioxidant, and Nitric Oxide (NO) Inhibitory Activities of Arbutus unedo L.

This study aims at investigating the contribution of two classes of compounds, flavonoids and iridoids, to the bioactivity of Arbutus unedo L. leaves and fruits. The impact of different extraction procedures on phytochemicals content and hypoglycemic, antioxidant, and nitric oxide (NO) inhibitory activities of A. unedo fresh and dried plant materials was investigated. Ellagic acid 4-O-ß-D-glucopyranoside, kaempferol 3-O-glucoside, and norbergenin were identified for the first time in this genus by using liquid chromatography-electrospray ionization-quadrupole-time of flight-mass spectrometry (LC-ESI-QTOF-MS). Three iridoids (gardenoside, geniposide, unedoside) are specifically identified in the leaves. Interestingly, asperuloside was extracted only from dried fruits by ethanol with Soxhlet apparatus. Extracts were screened for their potential antioxidant activities by using the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), Ferric Reducing Activity Power (FRAP), and ß-carotene bleaching tests. Based on the Global Antioxidant Score (GAS) calculation, the most promising antioxidant extract was obtained by hydroalcoholic maceration of dried leaves that showed half maximal inhibitory concentration (IC50) of 0.42 and 0.98 µg/mL in ABTS and DPPH assays, respectively. The hypoglycaemic activity was investigated by α-amylase and α-glucosidase inhibition tests. Extracts obtained by ethanol ultrasound extraction of fresh leaves and hydroalcoholic maceration of fresh fruits (IC50 of 19.56 and 28.42 µg/mL, respectively) are more active against α-glucosidase than the positive control acarbose (IC50 of 35.50 µg/mL). Fruit extracts exhibited the highest anti-inflammatory activity.

Validation and evaluation of four sample preparation methods for the quantification of intracellular tacrolimus in peripheral blood mononuclear cells by UHPLC-MS/MS.

Rejection and toxicity occur despite monitoring of tacrolimus blood levels during clinical routine. The intracellular concentration in lymphocytes could be a better reflection of the tacrolimus exposure. Four extraction methods for tacrolimus in peripheral blood mononuclear cells were validated and evaluated with UHPLC-MS/MS. Methods based on protein precipitation (method 1), solid phase extraction (method 2), phospholipids and proteins removal (method 3) and liquid-liquid extraction (method 4) were evaluated on linearity, lower limit of quantification (LLOQ), imprecision and bias. Validation was completed for the methods within these requirements, adding matrix effect and recovery. Linearity was 0.126 (LLOQ)-15 µg/L, 0.504 (LLOQ)-15 µg/L and 0.298 (LLOQ)-15 µg/L with method 1, 2 and 3, respectively. With method 4 non-linearity and a LLOQ higher than 0.504 µg/L were observed. Inter-day imprecision and bias were ≤4.6%, ≤10.9%; ≤6.8%, ≤-11.2%; ≤9.4%, ≤10.3% and ≤44.6%, ≤23.1%, respectively, with methods 1, 2, 3 and 4. Validation was completed for method 1 and 3 adding matrix effect (7.6%; 15.0%) and recovery (8.9%; 10.8%), respectively. The most suitable UHPLC-MS/MS method for quantification of intracellular tacrolimus was protein precipitation due to the best performance characteristics and the least time-consuming rate and complexity.