Antinociceptivna i protuupalna svojstva vodeno-etanolnog ekstrakta pokožice grožđa vrste Vitis labrusca izolirane iz otpada vinske industrije

Research background. Extracts from grape pomace, including the wine, show many biological effects such as antioxidant and anti-inflammatory activities. Unfortunately, winemakers discard the bagasse, so the waste is not exploited, although it contains bioactive compounds with antioxidant and anti-inflammatory properties. The work aims to analyze the hydroethanolic extract of peels from Vitis labrusca agro-industrial waste and to evaluate its antinociceptive and anti-inflammatory properties. This study is relevant for reusing a residue and adding value to the grape economic chain. Experimental approach. A representative sample of pomace was obtained and the peels were used to produce the extract. The phenolic compounds were determined by mass spectrometry in multiple reaction monitoring mode and Folin-Ciocalteu colorimetric method, using gallic acid as standard. The biological analyses were carried out using mice orally treated with crude extract at doses of 30, 100 and 300 mg/kg. We evaluated mechanical hyperalgesia by the von Frey method, thermal heat hyperalgesia using a hot plate at 55 °C, paw edema using a pachymeter, and neutrophil recruitment by measurement of myeloperoxidase activity. The nephrotoxicity and hepatotoxicity were evaluated by biochemical analyses using blood samples that were collected after the Vitis labrusca administration. Results and conclusions. In all wet winemaking residues peel mass fraction was 75%, and in dry residues 59%. We identified nine anthocyanins (3-O-glucosides: peonidin, delphinidin, petunidin and malvidin; 3-p-coumaroyl-glucosides: cyanidin, peonidin, petunidin and malvidin, and malvidin-3,5-diglucoside), five flavonoids (apigenin-7-glucoside, luteolin-7-glucoside, quercetin-3-galactoside, isorhamnetin-3-glucoside and myricetin-3-rutinoside), and mass fraction of phenolic compounds, expressed as gallic acid equivalents, was 26.62 mg/g. In vivo assays showed that Vitis labrusca extract at mass fractions 100 and 300 mg/kg reduced carrageenan-induced mechanical and thermal hyperalgesia, 50% of the paw edema, and neutrophil recruitment. In addition, there were no indications of nephrotoxicity and hepatotoxicity. Our extract obtained from winemaking residue has analgesic and anti-inflammatory properties, related at least in part to the presence of phenolic compounds, and it is not toxic to renal and hepatic tissues. Novelty and scientific contribution. This bio-product can be used as an alternative to synthetic anti-inflammatory agents with the same pharmacological potential and fewer side effects. We demonstrated that Vitis labrusca winemaking waste can be used for the production of antinociceptive and anti-inflammatory products (nutraceutical, pharmaceutical and cosmetics) without toxicity, contributing to the environmental economy.Pozadina istraživanja. Ekstrakt komine grožđa, kao i vino, ima mnoga biološka svojstva, poput antioksidacijskog i protuupalnog učinka. Nažalost, proizvođači vina odbacuju neiskorišteni trop iako sadržava bioaktivne spojeve s antioksidacijskim i protuupalnim svojstvima. Svrha je ovoga rada bila ispitati antinociceptivna i protuupalna svojstva vodeno-etanolnog ekstrakta pokožice grožđa vrste Vitis labrusca iz agroindustrijskog otpada. Značaj je ovoga istraživanja u tome što se korištenjem otpada daje dodatna ekonomska vrijednost grožđu u uzgojnom lancu. Eksperimentalni pristup. Ekstrakt je dobiven iz pokožica izdvojenih iz reprezentativnog uzorka komine. Udjel fenolnih spojeva određen je praćenjem višestrukih reakcija pomoću masene spektrometrije i metodom Folin-Ciocalteu, uz galnu kiselinu kao standard. Biološka aktivnost ekstrakata ispitana je na miševima koji su hranjeni sirovim ekstraktom u dozama od 30, 100 i 300 mg/kg. Ispitani su sljedeći parametri: mehanička hiperalgezija pomoću von Frey filamenata, toplinska hiperalgezija na vrućoj ploči pri 55 °C, edem šape pomoću pomičnog mjerila i aktivnost mijeloperoksidaze kao pokazatelj aktivacije neutrofila. Nefrotoksičnost i hepatotoksičnost su ispitane biokemijskim pretragama uzoraka krvi miševa hranjenih ekstraktom grožđa vrste Vitis labrusca. Rezultati i zaključci. Maseni udjel pokožica u svim uzorcima otpada vinske industrije iznosio je 75 % mokre tvari i 59 % suhe tvari. Identificirali smo devet različitih antocijanina (3-O-glukozide peonidin, delfinidin, petunidin i malvidin; 3-p-kumaroil-glukozide cijanidin, peonidin, pe¬tunidin i malvidin, te malvidin-3,5-diglukozid), pet flavonoida (apigenin-7-glukozid, luteolin-7-glukozid, kvercetin-3-galaktozid, izorhamnetin-3-glukozid i miri¬cetin-3-rutinozid), a maseni udjel fenolnih spojeva, izražen kao ekvivalent galne kiseline, bio je 26,62 mg/g. Ispitivanja in vivo pokazala su da su ektrakti grožđa vrste Vitis labrusca masenog udjela 100 i 300 mg/kg smanjili mehaničku i toplinsku hiperalgeziju kod miševa nakon injekcije karagenana, reducirali edem šape za 50 % i smanjili broj neutrofila. Osim toga, nije bilo pokazatelja nefrotoksičnosti i hepatotoksičnosti. Ekstrakt dobiven iz otpada vinske industrije ima analgetska i protuupalna svojstva, djelomično zbog toga što sadržava fenolne spojeve, a nije toksičan za tkiva bubrega i jetre. Novina i znanstveni doprinos. Dobiveni se bioproizvod može upotrijebiti kao alternativa sintetičkim protuupalnim agensima, s istim farmakološkim potencijalom a manje nuspojava. Pokazali smo da se vinski otpad grožđa vrste Vitis labrusca može upotrijebiti za proizvodnju antinociceptivnih i protuupalnih proizvoda (nutraceutičkih, farmaceutskih i kozmetičkih) koji nemaju toksični učinak, te na taj način pridonijeti zaštiti okoliša

The Lipid Mediator Resolvin D1 Reduces the Skin Inflammation and Oxidative Stress Induced by UV Irradiation in Hairless Mice

UV irradiation-induced oxidative stress and inflammation contribute to the development of skin diseases. Therefore, targeting oxidative stress and inflammation might contribute to reduce skin diseases. Resolvin D1 (RvD1) is a bioactive metabolite generated during inflammation to actively orchestrate the resolution of inflammation. However, the therapeutic potential of RvD1 in UVB skin inflammation remains undetermined, which was, therefore, the aim of the present study. The intraperitoneal treatment with RvD1 (3-100 ng/mouse) reduced UVB irradiation-induced skin edema, myeloperoxidase activity, matrix metalloproteinase 9 activity, and reduced glutathione depletion with consistent effects observed with the dose of 30 ng/mouse, which was selected to the following experiments. RvD1 inhibited UVB reduction of catalase activity, and hydroperoxide formation, superoxide anion production, and gp91phox mRNA expression. RvD1 also increased the Nrf2 and its downstream targets NQO1 and HO-1 mRNA expression. Regarding cytokines, RvD1 inhibited UVB-induced production of IL-1β, IL-6, IL-33, TNF-α, TGF-β, and IL-10. These immuno-biochemical alterations by RvD1 treatment had as consequence the reduction of UVB-induced epidermal thickness, sunburn and mast cell counts, and collagen degradation. Therefore, RvD1 inhibited UVB-induced skin oxidative stress and inflammation, rendering this resolving lipid mediator as a promising therapeutic agent

Optimization of ultrasound‐assisted extraction of grape‐seed oil to enhance process yield and minimize free radical formation

Grape seeds are a relatively abundant source of oil and bioactive compounds. To use this byproduct, the current work aimed to optimize the ultrasound‐assisted extraction (UAE) of grape‐seed oil to obtain greater process yield and minimize free radical formation in the oil. RESULTS The optimal condition was 15 °C with an ultrasonic wave amplitude of 42 µm, leading to a process yield of 82.9% and content of free radicals of 14.7 × 1017 kg−1 and 3.4 × 1018 kg−1 for samples stored for 7 and 30 days, respectively. No significant differences in fatty acid composition and acidity and iodine values were observed between samples. The oil obtained by ultrasound had greater phenolic compound content and antioxidant activity by ferric reduction than the control sample (without ultrasound application). However, higher content of free radicals and peroxide value was observed. CONCLUSION Sonication improved extraction yield when compared to the process without ultrasound application. Moreover, UAE favored the extraction of phenolic compounds. As it enhanced process yield with the minimum formation of free radicals, UAE is a promising oil‐extraction technology981350195026CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESSem informaçãoSem informaçã

Influence of vehicle on antioxidant activity of Quercetin: a liquid crystalline formulation

El efecto de una formulación líquido cristalina en la actividad antioxidante in vitro de la quercetina fue investigada utilizando diferentes metodologías con objetivo de escoger el método más adecuado para hacer el control de calidad y estudios de la estabilidad de la actividad antioxidante de la formulación líquido cristalina adicionada de la quercetina. Para todos los métodos empleados, la quercetina presentó actividad dosis dependiente con IC50 de 0.215 μg/mL en la inhibición de la peroxidación lípida, 1.125 μg/mL en la inhibición de la quimioluminiscência generada por el sistema H2O2 -HRP-luminol y 0.834 μg/mL en el ensayo de DPPH• y la actividad antioxidante de la quercetina adicionada a la formulación líquido cristalina fue valorada con suceso en todas las metodologías empleadas, ya que no hubo interferencia de los otros componentes da formulación y todas las metodologías se mostraron sensibles para evaluación de la quercetina. Así, la formulación líquido cristalina conteniendo vitamina E TPGS/ MIP / PG-H2 0 (1:1) en la porcentaje de 63.75/21.25/15 (p/p/p), respectivamente, pode ser potente para administración cosmética o farmacéutica, una vez que preservó el efecto antioxidante de la quercetina y todas las metodologías empleadas pueden ser utilizadas en el control de calidad y estudios de estabilidad de la actividad antioxidante da formulación líquido cristalina contenido quercetinaThe effect of a liquid crystalline formulation in the in vitro antioxidant activity of quercetin was investigated using different methodologies in order to choose the most suitable method to perform the quality control and stability studies of the antioxidant activity of the liquid crystalline formulation added with quercetin. For all the assays, quercetin showed a dose-dependent activity having an IC50 of 0.215 μg/mL in inhibiting lipid peroxidation, 1.125 μg/mL in inhibiting chemiluminescence produced in the H2O2 /luminol/horseradish peroxidase (HRP) system and 0.834 μg/mL in DPPH• assay and the antioxidant activity of quercetin in liquid crystalline formulation was successfully evaluated in all the methodologies applied, once no interference by other components of the formulation was found and all the methodologies were sensitive for the quercetin evaluation. Thus, the liquid crystalline formulation containing vitamin E TPGS/ IPM / PG-H2 0 (1:1) in the percentage of 63.75/21.25/15 (w/w/w), respectively, might be potential for cosmetic and pharmaceutical use, once it preserved the antioxidant effect of quercetin and all the methodologies applied could be used to perform the quality control and stability studies of the antioxidant activity of the liquid crystalline formulation added with quercetin.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Assessment of the antioxidant activity of two plant extracts containing isoflavonoids by different in vitro methods

Las especies reactivas del oxígeno (EROS) han sido relacionadas con diferentes enfermedades. Las EROS inducen daños y perjuicios a las membranas celulares vía la peroxidación lipídica (LPO), lo que contribuye a la oxidación del ADN y carcinogénesis. En ese sentido, varios estudios se centralizaron en la actividad anti-cancerosa de isoflavonoides presentes en la soja y en el trébol rojo. Sin embargo, para alcanzar su efecto terapéutico y permitir su amplia utilización, todo depende de la regularización y calidad de los extractos. En este estudio dos extractos comerciales, Isoflavin Beta ® (mezcla del isoflavonoides) y el extracto de trébol rojo, fueron caracterizados teniendo en cuenta sus constituyentes (flavonoides totales, isoflavonoides, polifenoles totales y proteínas totales). Además, sus actividades antioxidantes también fueron evaluadas por diferentes métodos. Los dos presentaron la actividad antioxidante significativa en todas las pruebas. Las concentraciones eficaces dependieron de cada prueba, lo que probablemente esté relacionado con sus constituyentes. Se concluiye que la composición química y la evaluación in vitro de la actividad antioxidante deben contribuir para la estandarización de los extractos de las plantas.The reactive oxygen species (ROS) may ultimately cause or participate in the induction of a wide range of diseases including cancer, atherosclerosis, rheumatoid arthritis and diabetes. Thus, many studies focus in the possible anticancerogenic activity of isoflavonoids present in soy and red clover. However, achieving therapeutic effect and a broader use depends on the extract standardization and quality. Therefore, in the present study two commercial extracts (Isoflavin Beta® - mixture of isoflavonoids and the dry red clover extract) were characterized on their constituents (total flavonoids, isoflavonoids, total polyphenols and total proteins), and in vitro antioxidant activities. Both extracts presented significant antioxidant activity in all tests. Moreover, the active doses depended on each test, probably because of extract composition. Concluding, chemical composition and in vitro antioxidant activity might help to standardize plant extracts.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Evaluation of the antioxidant activity as an additional parameter to attain the functional quality of natural extracts

Debido a las diferencias en la calidad funcional de extractos naturales, nosotros hemos advertido también diferencias en su efectividad, por lo que se pretende estimar la actividad antioxidante de extractos naturales para lograr su calidad funcional . Fue observado que todos los extractos (propolis marrón y verde, Ginkgo biloba e Isoflavin Beta®) y el patrón usado (quercetina) mostraron actividad antioxidante de una manera dosis-dependiente con valores de IC que van de 0,21 a 155,28 al µg mL (inhibición de la peroxidación lipídioca y captación de radicales libres del DPPH ). Se observó una alta correlación (r = 0,9913) entre los métodos antioxidantes y por otro lado la actividad antioxidante no estuvo relacionada con el contenido del polifenoles ni de flavonoides. Como el análisis del DPPH es un método rápido, presenta costos bajos e incluso tiene una correlación alta con otros métodos antioxidantes, este método podría utilizarse como un parámetro adicional en el control de calidad de extractos naturales.Due to differences in the functional quality of natural extracts, we have also faced differences in their effectiveness. So, it was intended to assess the antioxidant activity of natural extracts in order to attain their functional quality. It was observed that all the extracts (brown and green propolis, Ginkgo biloba and Isoflavin Beta®) and the standard used (quercetin) showed antioxidant activity in a dose-dependent manner with IC values ranging from 0.21 to 155.28 µg mL (inhibition of lipid peroxidation and scavenging of the DPPH assays). We observed a high correlation (r = 0.9913) among the antioxidant methods; on the other hand, the antioxidant activity was not related to the polyphenol and flavonoid content. As the DPPH assay is a fast method, presents low costs and even has a high correlation with other antioxidant methods, it could be applied as an additional parameter in the quality control of natural extracts.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Photostability of Quercetin under exposure to UV irradiation

Recientemente fue demostrado que formulaciones tópicas que contienen quercetina inhibieron la tensión oxidante y la inflamación cutánea inducida por la radiación UVB. No obstante, no fue aún investigada la fotoestabilidad de este flavonoide bajo la exposición a la radiación de UV y es conocido que la estabilidad fotoquímica de esta sustancia es esencial para su actividad. Por consiguiente, en este trabajo se investigó la fotoestabilidad de una solución de quercetina en propilenglicol bajo exposición a radiación UVB. Fue analizado el contenido de quercetina en cada solución por CLAR y su absorción al ultravioleta y su actividad antioxidante por DPPH• •Se validaron los métodos CLAR y de DPPH . y los resultados demostraron que estas metodologí- as son adecuadas y confiables para cuantificación y determinación de la actividad antioxidante de la quercetina, respectivamente. El estudio de fotoestabilidad mostró que la quercetina es resistente a la degradación causada por la exposición a la radiación UVB.Recently, it was demonstrated that topical formulations containing quercetin were able to inhibit the UVB-induced cutaneous oxidative stress and inflammation. Nevertheless, the photostability of this flavonoid under exposure to UV irradiation have not been investigated and it is well-known that the photochemical stability of this substance is essential for its activity. Therefore, in the present work was investigated the photostability of a propylene glycol solution of quercetin under forced exposure to UVB irradiation. The content of quercetin in each solution was determined quantitatively by HPLC, its ultraviolet spectra (UV-vis) and its antioxidant activity by DPPH• •The HPLC and DPPH . methods were validated and the results demonstrated that these methodologies are adequate and reliable to quantify quercetin and to determine its antioxidant activity, respectively. The photostability study showed that quercetin is resistant of degradation caused by UVB radiation exposure.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Method validation and stability study of quercetin in topical emulsions

This study validated a high performance liquid chromatography (HPLC) method for the quantitative evaluation of quercetin in topical emulsions. The method was linear within 0.05 - 200 μg/mL range with a correlation coefficient of 0.9997, and without interference in the quercetin peak. The detection and quantitation limits were 18 and 29 ng/mL, respectively. The intra- and inter-assay precisions presented R.S.D. values lower than 2%. An average of 93% and 94% of quercetin was recovered for non-ionic and anionic emulsions, respectively. The raw material and anionic emulsion, but not non-ionic emulsion, were stable in all storage conditions for one year. The method reported is a fast and reliable HPLC technique useful for quercetin determination in topical emulsions