Intensification of marrubiin concentration by optimization of microwave-assisted (low CO2 yielding) extraction process for Marrubium vulgare using central composite design and antioxidant evaluation

Context: Marrubium vulgare Linn (Lamiaceae) was generally extracted by conventional methods with low yield of marrubiin; these processes were not considered environment friendly.

Objective: This study extracts the whole plant of M. vulgare by microwave assisted extraction (MAE) and optimizes the effect of various extraction parameters on the marrubiin yield by using Central Composite Design (CCD).

Materials and methods: The selected medicinal plant was extracted using ethanol: water (1:1) as solvent by MAE. The plant material was also extracted using a Soxhlet and the various extracts were analyzed by HPTLC to quantify the marrubiin concentration.

Results: The optimized conditions for the microwave-assisted extraction of selected medicinal plant was microwave power of 539?W, irradiation time of 373?s and solvent to drug ratio, 32?mL per g of the drug. The marrubiin concentration in MAE almost doubled relative to the traditional method (0.69?±?0.08 to 1.35?±?0.04%). The IC₅₀ for DPPH was reduced to 66.28?±?0.6?μg/mL as compared to conventional extract (84.14?±?0.7?μg/mL). The scanning electron micrographs of the treated and untreated drug samples further support the results.

Discussion and conclusion: The CCD can be successfully applied to optimize the extraction parameters (MAE) for M. vulgare. Moreover, in terms of environmental impact, the MAE technique could be assumed as a ‘Green approach’ because the MAE approach for extraction of plant released only 92.3?g of CO₂ as compared to 3207.6?g CO₂ using the Soxhlet method of extraction.     

The influence of extraction methods on rutin yield of cassava leaves (Manihot esculenta Crantz)

Rutin, a well-known bioflavonoid, was found abundantly in cassava leaves. In the present study, extraction techniques including maceration, boiling, reflux, ultrasound-assisted extraction (UAE), and microwave-assisted extraction (MAE) were optimised to increase the yield of rutin. Extraction parameters such as solvents, solid-liquid ratio, temperature, and time were optimised to give better extraction yields for each method. HPLC analysis showed a high content of rutin which is up to 2.4% per dry weight of cassava leaves. The extraction yields under optimised condition were found to be 16.00 ± 0.21, 20.38 ± 0.66, 22.33 ± 2.3, 24.49 ± 0.41, and 23.37 ± 1.00 g rutin per kg dry weight for maceration, boiling, reflux, UAE and MAE methods, respectively. Specifically, UAE reduced the extraction time to 90 min, using only 40–60% of aqueous ethanol. Meanwhile, MAE completed the extraction under 5 min and no significant differences in output was observed between the use of water and aqueous ethanol. Accordingly, with the extraction efficiency of up to 99 and 94%, respectively, both processes provided better results. The subsequent green purification using chilling method produced a typical cassava bioflavonoid containing 82% of rutin and 17% of nicotiflorin. This study informs a new abundant source of rutin and provides the optimum condition of extraction methods for high yield of rutin from cassava leaves.     

Microwave hydrodiffusion and gravity model with a blend of high and low power microwave firing for improved yield of phenolics and flavonoids from oyster mushroom

Oyster mushrooms are a globally important entity both commercially and therapeutically with immense food value. The work describes a successful application of a unique method of microwave hydrodiffusion and gravity combining optimal mix of high and low power microwave firing for the extraction of phenolics and flavonoids principles from oyster mushroom for the first time. Initial firing of microwaves at higher power level (510 W for 2 min and 340 W for 2 min) was applied followed by sustained microwave firing at 170 W till the completion of the extraction as indicated by physical oozing out of the aqueous extract. Such a combination of microwave firing in microwave hydrodiffusion and gravity (MHG) method (26 min) successfully produced two times more yield of extract with 57.7% ± 2.8 and 82.3% ± 3.7 more content of phenolics and flavonoids, respectively, when compared to 5 h Soxhlet extraction and that too with a better reproducibility. Mapping of individual phenolics/flavonoids were carried out and SEM images were studied for better understanding of the operational aspects. The research aims towards canvassing green technologies as in the near future only those technologies shall survive which are in tandem with environment.     

Extraction of bioactive compounds from buriti (Mauritia flexuosa L.) fruit by eco-friendly solvents: Chemical and functional characterization

Buriti (Mauritia flexuosa L.) is a palm tree found in several regions of Latin America. Buriti fruit is a rich source of bioactive compounds such as carotenoids and phenolic compounds. Thus, the aim of this study was to extract bioactive compounds from buriti fruit by ethanol and a supramolecular solvent system (SUPRAS) formed by octanoic acid aggregates. The extracts were evaluated for total carotenoids, β-carotene, phenolic compounds and antioxidant capacity. Additionally, SUPRAS extracts were characterized for antibacterial activity and modulating effect. The extraction of β-carotene with SUPRAS showed a yield of 5.82 ± 0.05 mg/g for the peel and 26.7 ± 0.02 mg/g for the pulp. In relation to total phenolic compounds, the yields were 32.1 ± 1.2 μg GAE/g for the peel and 24.53 ± 4.9 μg GAE/g for the pulp. The presence of gallic acid, quercetin and catechin stand out regarding the phenolics identified. The extracts showed antioxidant activity, with an emphasis on the extracts obtained by SUPRAS, which presented EC₅₀ (concentration required to obtain a 50% antioxidant effect) for the ABTS radical sequestration of 3.00 μg/mL for the peel and 0.84 μg/mL for the pulp. When combined with norfloxacin and gentamicin antibiotics, the extracts also showed a synergistic action against multi-resistant strains of Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli. Thus, the extraction of bioactive compounds from buriti fruit using a safe, biocompatible, biodegradable and environmentally friendly solvent such as SUPRAS represents potential for developing new pharmaceuticals, cosmetics and functional foods.     

Economic pre-feasibility of supercritical fluid extraction of antioxidants from fruit residues

During the different stages of the supply chain, large amounts of waste are generated, being mostly disposed of in landfills. Most of these residues have a high content of valuable compounds with a high antioxidant capacity. These compounds have a high potential for use in different industries, such as the pharmaceutical and food industries. Supercritical fluid extraction is one of the technologies available for extracting effectively these components from wastes. Several studies have focused on this technology to show its advantages. This work aims evaluates the production of bioactive compound from different fruit residues to analyze their economic pre-feasibility using supercritical fluid extraction. Five fruit wastes were used for the analysis: mango peel, yellow passion fruit seed, raspberry seeds, mandarin peel, and açaí berry exhausted pulp. As results there was found that the use of a co-solvent increases production costs. Another influencing factor was the extraction yield related to the raw material used. The appropriate selection of these variables makes it possible to obtain economically viable processes. The yellow passion fruit presents the better economic indicators for obtaining an oil extract mainly composed of palmitic acid. With the raw material flow of 100 kg/h, a profit margin of 86.94% was obtained, which was the higher than for other raw materials. Additionally, the minimum flow of yellow passion fruit is 5.13 kg/h to obtain an NPV of zero at ten years.     

Optimization of microwave-assisted extraction of solanesol from potato leaves and stems

Microwave-assisted extraction (MAE) was developed for the fast extraction of solanesol from potato leaves and stems. The ratio of raw material to ethanol, extraction time, extraction temperature, and microwave irradiation power were interdependent. The yield of solanesol reached its maximum 98.57% with 1:8.0 g/ml, 40 min, 55°C, and 2.0 KW, respectively. MAE was comparable to other extraction methods, including solvent extraction, heat-reflux, and Soxhlet-extraction. MAE reduced extraction time, solvent consumption, and increased yields of solanesol. Solanesol concentration was performed by RP-high performance liquid chromatography. The method was rapid, simple, accurate, and reproducible.     

Green extraction of Buchanania siamensis and water-based formulations

Buchanania siamensis is a local plant in eastern and north-eastern Thailand belonging to the Anacardiaceae family. The plant provides shading and is a good source of food and medicine since its fruits possess antioxidant and anti-cancer effects. The green, environmental-friendly, extraction of B. siamensis, under ultrasound- and microwave-assisted extraction, using solvents consisting of a variation of tween 80, propylene glycol (PG), and ethanol concentrations were investigated. Extraction by aqueous PG under ultrasound-assisted extraction (UAE) conditions yielded the highest total phenolic (TPC) and total flavonoid contents (TFC). The optimum condition, which was using a 40 mL/g ratio, 50% PG at 60°C for 20 min, yielded the extract with TPC at 49.12 ± 0.51 mgGAE/g DW and TFC at 14.18 ± 0.48 mgQE/g DW. The extract exhibited antioxidant activity at IC₅₀ 2.49 ± 0.03 mg/mL. Water-based formulations (solution and hydrogel) of this extract were successfully fabricated. The formulations demonstrated stable properties, TPC, myriscitrin, quercetrin content, and antioxidant properties after the heating and cooling test (6 cycles). Consequently, the B. siamensis extract, rich in antioxidative agents, was proper for incorporation into water-based cosmeceutical products.     

Recovery of phenolic compounds from spent coffee grounds through optimized extraction processes

The aim of the present study was to compare novel methods and technologies of phenolics extraction from spent coffee grounds, as “green” alternative techniques of the conventional ethanol extraction. Three extraction techniques, ultrasound- and microwave-assisted extraction, along with the use of β-cyclodextrin as a solvent have been studied and optimized as the eco-friendly solutions for the efficient extraction of phenolic compounds from Spent Coffee Grounds (SCG). The investigated factors were temperature (20–60 °C), solvent concentration (0–100% v/v water ethanol, EtOH), amplitude (20–60%), and liquid/solid ratio (5–60 mL/g) for ultrasound-assisted extraction (UAE). In the case of microwave extraction (MAE), the effects of solvent concentration (0–100% v/v), liquid/solid ratio (5–60 mL/g), and power (100–600 Watt) were studied. Ethanol (0–100% v/v) and β-cyclodextrin (β-CD, 1–18.5 mg/mL) were also compared as solvents, under the same range of liquid/solid ratio (5–60 mL/g) and temperatures (20–60 °C). In addition, the effect of the drying method (oven drying and freeze-drying), as well as a defatting pretreatment of SCG, were studied under the optimum conditions of each extraction method. The optimum extraction yield (31.79 ± 0.25 mg GAE/g SCG) was achieved using MAE with a liquid/solid ratio of 60 mL/g, a power level 600 W, and a solvent concentration of 68% v/v EtOH. While the maximum yield (34.43 mg GAE/g SCG) was also obtained by the same method from the freeze-dried sample.     

Comparing ultrasonic- and microwave-assisted methods for extraction of phenolic compounds from Kabkab date seed (Phoenix dactylifera L.) and stepwise regression analysis of extracts antioxidant activity

This study aimed to extract bioactive compounds (phenolics and flavonoids and condensed tannins) from roasted date seed (Phoenix dactylifera L. cv Kabkab) using various solvent systems (W: water, AE: aqueous-ethanol, AA: aqueous-acetone) and extraction method (ultrasonic-assisted (UAE), microwave-assisted (MAE) and the combination of these two methods (UMAE) maceration (ME) and Decoction-Infusion (DIE) Extraction). Moreover, the feasibility of antioxidant activity prediction was investigated based on stepwise regression analysis and phytochemical properties. Extraction yield, Total phenolic content (TPC), total flavonoid content (TFD) and antioxidant activity (DPPH*, ABTS*⁺, FRAP and averaged antioxidant activity: AAA) of the extracts were evaluated. The main effect of solvent systems and extraction methods on phytochemical compounds and antioxidant activity were significant (P < 0.01). Water and aqueous-ethanol solvents extracted higher phytochemical compounds than aqueous-acetone (P < 0.05). Although the highest extraction performance was observed for the ME method, the novel methods show an acceptable result in a much shorter time. Among novel methods, the highest and lowest performances were recorded for UAE and MAE. There was no significant difference between novel-green methods (e.g., UAE and UMAE). Although the lowest phytochemical yield was obtained for MAE, this performance was obtained in less than 5 min. The highest and lowest correlation between the phytochemical compositions and antioxidant activity parameters were for DPPH* and AAA, respectively. Stepwise-regression analysis showed the weakest and strongest prediction models for AAA (10-fold S = 0.098 and 10-fold R² = 87.07) and ABTS*⁺ (10-fold S = 0.129 and 10-fold R² = 78.25), respectively.     

Use of generally recognized as safe or dietary compounds to inhibit buprenorphine metabolism: potential to improve buprenorphine oral bioavailability

The present study evaluated the potential of five generally recognized as safe (GRAS) or dietary compounds (α‐mangostin, chrysin, ginger extract, pterostilbene and silybin) to inhibit oxidative (CYP) and conjugative (UGT) metabolism using pooled human intestinal and liver microsomes. Buprenorphine was chosen as the model substrate as it is extensively metabolized by CYPs to norbuprenorphine and by UGTs to buprenorphine glucuronide. Chrysin, ginger extract, α‐mangostin, pterostilbene and silybin were tested for their inhibition of the formation of norbuprenorphine or buprenorphine glucuronide in both intestinal and liver microsomes. Pterostilbene was the most potent inhibitor of norbuprenorphine formation in both intestinal and liver microsomes, with IC₅₀ values of 1.3 and 0.8 μM, respectively, while α‐mangostin and silybin most potently inhibited buprenorphine glucuronide formation. The equipotent combination of pterostilbene and ginger extract additively inhibited both pathways in intestinal microsomes. Since pterostilbene and ginger extract showed potent CYP and/or UGT inhibition of buprenorphine metabolism, their equipotent combination was tested to assess the presence of synergistic inhibition. However, because the combination showed additive inhibition, it was not used while performing IVIVE analysis. Based on quantitative in vitro–in vivo extrapolation, pterostilbene (21 mg oral dose) appeared to be most effective in improving the mean predicted F_oral and AUC_∞^PO of buprenorphine from 3 ± 2% and 340 ± 330 ng*min/ml to 75 ± 8% and 36,000 ± 25,000 ng*min/ml, respectively. At a 10‐fold lower dose of pterostilbene, the predicted buprenorphine F_oral approximated sublingual bioavailability (~35%) and showed a 2–4 fold reduction in the variability around the predicted AUC_∞^PO of buprenorphine. These results demonstrate the feasibility of using various GRAS/dietary compounds to inhibit substantially the metabolism by CYP and UGT enzymes to achieve higher and less variable oral bioavailability. This inhibitor strategy may be useful for drugs suffering from low and variable oral bioavailability due to extensive presystemic oxidative and/or conjugative metabolism.     

Stability,permeation, and cytotoxicity reduction of capsicum extract nanoparticles loaded hydrogel containing wax gourd extract

The aim of this study was to develop hydrogel loaded with capsicum extract nanoparticles and wax gourd extract for transdermal delivery of capsaicin. The addition of wax gourd extract was supposed to reduce cytotoxicity of capsaicin in capsicum extract against HaCaT keratinocyte cell line. Capsicum extract nanoparticles were prepared by solvent displacement method using hyaluronic acid as a stabilizer. The physical and chemical stability of capsicum extract nanoparticles were investigated by dynamic light scattering technique and UV–Visible spectrophotometry, respectively. Hydrogel loaded with capsicum extract nanoparticles and wax gourd fruit extract was then formulated by using Carbopol 940® as a gelling agent for transdermal delivery. The skin permeability of capsaicin from the hydrogel was evaluated by Franz diffusion cell approach. The cytotoxicity reduction of capsicum extract nanoparticles and capsicum extract nanoparticles by mixing with wax gourd extract was determined by MTT assay The results showed that capsicum extract nanoparticles exhibited an average diameter of 168.4 ± 5.3 nm with a polydispersity index and zeta potential value of 0.26 ± 0.01 and −45.7 ± 7.1 mV, respectively. After two month-storage, particle size, polydispersity index, and zeta potential values of capsicum extract nanoparticles stored at 4° C, 30° C, and 45 °C did not significantly change. The capsaicin content decreased to 78%, 71%, and 72% when stored at 4 °C, 30 °C, and 45 °C for three months, respectively. The pH values of hydrogel containing capsicum extract nanoparticles were found to be in the range of 5.58–6.05 indicating good stability. The hydrogel exhibited a pseudoplastic character. The rate of permeation flux of capsaicin from hydrogel was 7.96 µg/cm²/h. A significant increase in cell viability was observed when the cells were incubated with capsicum extract nanoparticles mixed with wax gourd, compared to capsicum extract nanoparticles alone. The wax gourd extract in the hydrogel protected HaCaT cells from capsaicin cytotoxicity, thus may provide a new approach for delivery of capsaicin to reduce cytotoxicity to skin cells.     

Ultrasonication and encapsulation of Butcher broom (Ruscus Hyrcanus L.) extract and its bioactive effects on qualitative properties,oxidative stability and shelf life of cake

Three solvents of pure-water (PW), 50:50 ethanol-water (EW) and pure-ethanol (PE) were used to obtain phytochemicals of Butcher's broom (Ruscus Hyrcanus L) leaves (BBL) separately with 10% concentrations (w/w) at 20 °C for 48 h. Since the extract yield of EW was higher than other solvents, therefore 100, 300, 500 and 700 mg/L of BBL were macerated separately at similar conditions and ultrasonicated in 37 kHz at 35 °C for 35 min. The yield of ultrasound assisted extraction (UAE) of EW had significantly (P ≤ 0.05) higher DPPH, ferric reducing antioxidant power (FRAP), phenolic, anthocyanin and flavonoids than EW extract before sonication at each BBL concentration. Then the yield of UAE at 700 mg/L was nano-capsulated (NC) with the mixture of (50:50) maltodextrin/gum Arabic (MD/GA). The NC efficiency and the coated-particle sizes were 80.48% and 95.17 nm, respectively. Furthermore, the scanning electron microscopy (SEM) and test of Fourier-transform infrared spectroscopy (FTIR) confirmed successful encapsulation and interactions between the final extract yield and coating layer of MD/GA. The UAE extracts of 700 mg/L with and without encapsulation were separately added into the prepared cake dough at 0.7%. Analysis of baked cakes during 45-day storage at 20 °C showed that thiobarbituric acid, peroxide values, yeast, mold and total viable count of NC-samples were significantly (P ≤ 0.05) lower than non-capsulated UAE-sample and control (cake without any extract). The MD/GA layer of NC-sample could protect the BBL phytochemicals and extend the shelf life of cake significantly longer (without negative quality indictors) due to its stable coating at 150 °C baking temperature.     

Valorisation of Salicornia ramosissima biowaste by a green approach – An optimizing study using response surface methodology

The purpose of this study was to optimize the extraction of antioxidants from Salicornia ramosissima by-product through Response Surface Methodology (RSM). The impacts of time (10–60 min) and temperature (40–80 °C) in total phenolic content (TPC) and antioxidant/antiradical activity were assessed. The optimal extract was attained at the binomial 80 °C/10 min and characterized regarding phenolic composition, pigments quantification, radicals quenching capacity, anti-acetylcholinesterase (AChE) activity, and cell viability effects. The optimal extract was characterized by low carotenoids content (8.15 μg/g on dry weight (dw)) and high amounts of hydroxycinnamic acid derivatives (23.12 mg/g dw). A remarkable scavenging efficiency was observed, particularly for HOCl (IC₅₀ = 27.61 μg/mL) and ONOO^? (IC₅₀ = 47.32 μg/mL). No adverse effects were detected on HaCaT and HFF-1 viability up to 1000 μg/mL. Additionally, the neuroprotective properties of the optimal extract were demonstrated by AChE inhibition (32.34% at 1000 μg/mL). This study encourages the sustainable recovery of antioxidants from the undervalued biowaste of S. ramosissima as a new raw material for different industries.     

Species specificity profiling of rat and human organic cation/carnitine transporter Slc22a5/SLC22A5 (Octn2/OCTN2)

The purpose of this study was to characterize the uptake of carnitine, the physiological substrate, and the uptake of 3-(2,2,2-trimethylhydrazinium)propionate, a consensus substrate by rat Octn2 and human OCTN2 transporters as well as to characterize drug-mediated inhibition of l-carnitine uptake by the rat and human orthologs overexpressed in CHO-K1 cells. l-carnitine and 3-(2,2,2-trimethylhydrazinium)propionate were found to be a lower affinity substrate for rat Octn2 (K_M = 32.66 ± 5.11 μM and 23.62 ± 4.99 μM respectively) than for human OCTN2 (K_M = 3.08 ± 0.74 μM and 7.98 ± 0.63 μM). The intrinsic clearance (CL_int) value for carnitine was higher for the human than for the rat transporter (22.82 ± 5.57 ml/min*mg vs 4.008 ± 0.675 ml/min*mg). For 3-(2,2,2-trimethylhydrazinium)propionate, in contrast, the CL_int value for rat Octn2 was higher than for human OCTN2 (323.9 ± 72.8 ml/min*mg vs 65.11 ± 5.33 ml/min*mg).Furthermore, many pharmacologically important drugs were shown to affect l-carnitine transport by Octn2/OCTN2. The correlation between the IC₅₀ datasets for the rat and human transporter resulted in an r value of 0.47 (p > 0.05). However, the greatest difference was less than seven-fold and 13 of 15 compounds yielded a difference less than 3-fold.Thus, the transporters from these two species showed an overlapping but somewhat different substrate and inhibitor specificity.     

Solid self-emulsifying nitrendipine pellets: Preparation and in vitro/in vivo evaluation

Objective of this study is to develop and evaluate the new solid self-emulsifying (SE) pellets of poorly soluble nitrendipine (NTD). These pellets were prepared via extrusion/spheronization technique, using liquid SEDDS (NTD, Miglyol^® 812, Cremophor^® RH 40, Tween 80, and Transcutol^® P), adsorbents (silicon dioxide and crospovidone), microcrystalline cellulose and lactose. The resulting SE pellets with 30% liquid SEDDS exhibited uniform size (800–1000 μm) and round shape, droplet size distribution following self-emulsification was nearly same to the liquid SEDDS (72 ± 16 nm and 64 ± 12 nm). The in vitro release was similar for the two SE formulations (over 80% within 30 min), both significantly higher than the conventional tablets (only 35% within 30 min). The oral bioavailability was evaluated for the SE pellets, liquid SEDDS and conventional tablets in fasted beagle dogs. AUC of NTD from the SE pellets showed 1.6-fold greater than the conventional tablets and no significant difference compared with the liquid SEDDS. In conclusion, our studies illustrated that extrusion/spheronization technique could be a useful large-scale producing method to prepare the solid SE pellets from liquid SEDDS, which can improve oral absorption of NTD, nearly equivalent to the liquid SEDDS, but better in the formulation stability, drugs leakage and precipitation, etc.     

Evaluation of green extraction methods on bioactive compounds and antioxidant capacity from Bougainvillea glabra bracts

Bougainvillea glabra is a native plant of South America, and its bracts are still an underexploited source of betalains and phenolic compounds. Microwave (MAE), ultrasound (UAE), conventional aqueous and ethanolic exhaustive extractions were used for the recovery of total phenolic compounds (TPC) and betalains from B. glabra bracts. The highest yields of TPC, betalains, and antioxidant capacity by ABTS were obtained by ethanolic exhaustive extraction followed by MAE at 100 and 600 W. Deoxyribose protection assay measurement by hydroxyl radical scavenging showed high values for extracts obtained by exhaustive extraction and MAE, ranging from 89% to 83%, indicating a high capacity to capture hydroxyl radicals. Agarose gel electrophoresis assay showed that the extracts obtained by UAE, conventional aqueous, and ethanolic extraction were able to protect DNA from free radicals and did not show pro-oxidant effect. Seventeen phenolic compounds and seventeen betalains were identified by HPLC/MS in the extract of B. glabra bracts obtained by exhaustive extraction, composed mainly of kaempferol derivatives (76.5%) and acylated betalains (71.0%), respectively. The results suggest that MAE may be an alternative to recover betalains and phenolic compounds from B. glabra bracts due to the lower extraction times and the absence of organic solvent.     

Characterization of a green expanded DGT methodology for the in-situ detection of emerging endocrine disrupting chemicals in water systems

Performances of sustainable and easy-to-make agarose (1.5%) – AC (1%) DGTs to monitor emerging EDCs were investigated. TGA, EDX showed agarose (1.5%) and agarose (1.5%) – AC (1%) biodegradable gels have constant water-carbon ratio along the two-dimensions ensuring consistent diffusive property and binding abilities. Gaussian simulations gave back lengths and widths of (12.32 Å; 5.11 Å), (12.32 Å; 6.77 Å), (10.18 Å; 4.36 Å), (10.23 Å; 4.83 Å) for E2, EE2, BPA and BPAF, respectively. FE-SEM characterization showed agarose (1.5%) has average pore number of 22 ± 5 μm² with d.nm Ø 30 nm, while agarose (1.5%) – AC (1%) 13 ± 2 μm² with d.nm Ø 45, ensuring free analytes diffusion. Effective diffusion coefficients at 25 °C were (3.97 ± 0.98 · 10-6 cm² s^-1), (3.18 ± 0.04 · 10-6 cm² s^-1), (3.09 ± 0.61 · 10^?6 cm² s^-1), (3.55 ± 1.03 · 10^-6 cm² s^-1) for E2, EE2, BPA and BPAF respectively. Kinetic studies showed agarose (1.5%) – AC (1%) uptakes, over the first 60 min, 97%, 95%, 98%, 98% of E2, EE2, BPA, BPAF, respectively with relative binding rates of 2.48, 3.55, 2.05, 1.20 ng cm^-2 min^-1. Agarose (1.5%) – AC (1%) DGT resulted suitable for EDCs in trace detection.     

Gastroprotective potential of methanolic extract and dimethyl cardamonin from Campomanesia reitziana fruits in mice

This study evaluated the gastroprotective potential of methanolic extract from fruits of Campomanesia reitziana (MECR) and its isolated chalcone dimethyl cardamonin (DMC). The phenolic compound in the extract, and the free radical scavenging activity of MECR and DMC, were quantified. The gastroprotective activity of MECR (30–300 mg/kg, p.o) and DMC (1 and 3 mg/kg, p.o) was determined by ethanol/HCl-induced gastric ulcers in mice. Histological, histochemical, and biochemical analyses were performed in the ulcerated tissue. MECR showed a high content of phenolic compounds, including DMC, and was able to scavenge DPPH radicals by 29.58% at 1000 μg/mL. However, DCM (1–1000 μg/mL) did not reduce DPPH radicals. Pre-treatment with MECR at doses of 100 and 300 mg/kg reduced the gastric lesions by 35.07 and 79.47%, respectively (ulcerated-vehicle group 10.72 ± 0.88 mm²). Moreover, the extract increased the mucin content by 1044.44% and superoxide dismutase activity by 20.04%, and decreased the lipoperoxide levels by 39.39%, compared to the ulcerated-vehicle group (0.27 ± 0.04 pixels × 10³/field; 57.37 ± 1.59 U SOD/mg of protein and 29.57 ± 2.99 mmol LOOH/mg of tissue, respectively). However, MECR did not prevent the depletion of reduced glutathione or the decrease in catalase activity. Pre-treatment with DMC, at 1 and 3 mg/kg, also reduced the gastric ulcers by 53.83 and 52.64%, respectively. In summary, these findings confirm the gastroprotective activity of MECR and DMC, and open an interesting field concerning the gastroprotective potential of dimethyl cardamonin, given its potent antinociceptive and anti-inflammatory activity already described.

     

Anti-inflammatory properties of fruit juices enriched with pine bark extract in an in vitro model of inflamed human intestinal epithelium: The effect of gastrointestinal digestion

Enrichment of fruit juices with pine bark extract (PBE) could be a strategy to compensate for phenolic losses during the gastrointestinal digestion. A coculture system with Caco-2 cells and RAW 264.7 macrophages was established as an in vitro model of inflamed human intestinal epithelium for evaluating the anti-inflammatory capacity of fruit juices enriched with PBE (0.5 g L⁻¹) before and after in vitro digestion. The digestion of both PBE-enriched pineapple and red fruit juice led to significant changes in most of the analysed phenolic compounds. The in vitro inflammatory state showed cell barrier dysfunction and overproduction of IL-8, nitric oxide (NO) and reactive oxygen species (ROS). In the inflamed cells, incubation with nondigested samples reduced (P < 0.05) the production of IL-8 and NO compared with digested samples. ROS production increased in the inflamed cells exposed to digested commercial red fruit juice (86.8 ± 1.3%) compared with fresh juice (77.4 ± 0.8%) and increased in the inflamed cells exposed to digested enriched red fruit juice (82.6 ± 1.6%) compared with the fresh enriched juice (55.8 ± 6%). The anti-inflammatory properties of PBE-enriched fruit juices decreased after digestion; further research on the bioavailability of the assayed compounds is needed to properly assess their usefulness for the treatment of gut inflammation.     

Glucuronidation of piceatannol by human liver microsomes: major role of UGT1A1, UGT1A8 and UGT1A10

Objectives Piceatannol, a dietary polyphenol present in grapes and wine, is known for its promising anticancer and anti‐inflammatory activity. The aim of this study was to analyse the concentration‐dependent glucuronidation of piceatannol in vitro. Methods To determine the glucuronidation of piceatannol, experiments were conducted with human liver microsomes as well as using a panel of 12 recombinant UDP‐glucuronosyltransferase isoforms. Furthermore, the chemical structures of novel glucuronides were identified by liquid chromatography‐tandem mass spectrometry (LC‐MS/MS). Key findings Along with piceatannol it was possible to identify three metabolites whose structures were identified by LC‐MS/MS as piceatannol monoglucuronides (M1–M3). Formation of M1 and M3 exhibited a pattern of substrate inhibition, with apparent K_i and V_max/K_m values of 103 ± 26.6 µm and 3.8 ± 1.3 µl/mg protein per min, respectively, for M1 and 233 ± 61.4 µm and 19.8 ± 9.5 µl/mg protein per min, respectively, for M3. In contrast, formation of metabolite M2 followed classical Michaelis–Menten kinetics, with a K_m of 18.9 ± 8.1 µm and a V_max of 0.21 ± 0.02 nmol/mg protein per min. Incubation in the presence of human recombinant UDP‐glucuronosyltransferases (UGTs) demonstrated that M1 was formed nearly equally by UGT1A1 and UGT1A8. M2 was preferentially catalysed by UGT1A10 and to a lesser extent by UGT1A1 and UGT1A8. The formation of M3, however, was mainly catalysed by UGT1A1 and UGT1A8. Conclusions Our results elucidate the importance of piceatannol glucuronidation in the human liver, which must be taken into account in humans after dietary intake of piceatannol.