Development of hydrosoluble gels with Crataegus monogyna extracts for topical application: evaluation of antioxidant activity of the final formulations

Crataegus monogyna, commonly known as hawthorn, is notorious for having different ethnopharmacological applications. The antioxidant activity of methanolic extracts from its flowers and fruits in several stages of development was formerly reported by us. Therefore it is believable that the extracts of its botanical parts might have great potential to be incorporated in innovative pharmaceutical formulations. Accordingly, several paraben free, carbopol 940 based semisolid formulations were prepared with different hawthorn parts extracts. The antioxidant properties of ethanolic and aqueous hawthorn extracts, as well as their corresponding hydrosoluble gels, were screened. Ethanolic extracts proved to be more effective, while flowers and unripe fruits were the most bioactive hawthorn parts. The prepared hydrosoluble gels, that presented an adequate consistency, a pleasant colour and a non greasy texture, kept almost entirely the antioxidant activity exhibited by the extracts from which they were prepared. In addition, the hydrosoluble gels were tested and promptly absorbed by the skin and did not show significant pH alterations during the 90 days of observation. Despite the need of further tests (viscosity, extrudability, spreadability or transepidermal water loss), the prepared formulations demonstrate interesting attributes to be explored as potential dermopharmaceutical products

Flores e frutos imaturos de Crataegus monogyna revelam elevado potencial antioxidante.

A espécie Crataegus monogyna, conhecida vulgarmente como espinheiro, escaramunheiro ou pilriteiro, está entre as plantas do Nordeste de Portugal mais recomendadas pela medicina tradicional. As suas bagas são comumente consumidas por serem consideradas nutritivas e saudáveis, podendo também ser utilizadas como suplemento alimentar, sobretudo para crianças, devido ao seu elevado conteúdo vitamínico [1]. O espinheiro é reconhecido como tendo propriedades bioativas de interesse elevado, em particular na prevenção de doenças cardiovasculares e gastrointestinais. De forma a compreender melhor o potencial terapêutico do espinheiro, a atividade antioxidante de diferentes partes da planta (botões e brácteas florais, flores e frutos) foi avaliada através de diferentes métodos químicos e bioquímicos. Os extratos etanólicos demonstraram maior potencial antioxidante que os extratos aquosos, mas os resultados foram muito satisfatórios em ambos os casos, revelando o potencial do espinheiro para ser incluído em formulações terapêuticas ou produtos dietéticos de referência

Evaluation of growth inhibitory activity of Crataegus monogyna Jacq. flower bud extracts against human tumor cell lines

The vast structural diversity of natural compounds found in plants provides unique opportunities for discovering new drugs with possible beneficial effects on human health, including regulation of proliferation and cell death pathways leading to cancer [1]. Crataegus monogyna Jacq. has been studied by our research group to confirm its different alleged health benefits [2-4]. Herein, the antiproliferative activity of phenolic extracts from C. monogyna flower buds was evaluated on four different human tumor cell lines: HeLa, cervical carcinoma; HepG2, hepatocellular carcinoma; MCF-7, breast adenocarcinoma; NCI-H460, non-small cell lung cancer. The antiproliferative activity was measured by calculating Gl50 values (50% of cell growth inhibition) for each cell line, according to the procedure adopted by the National Cancer Institute for in vitro anticancer drug screening, which uses sulforhodamine B assay to assess cell groW1h inhibition [5]. The obtained antiproliferative activity was very similar among the assayed cell lines: HeLa, Gl50 = 63.55±3.56 μg/ml; HepG2, Gl50 = 88.45±8.11 μg/ml; MCF7, Gl50 = 66.96±0.01 μg/ml ; NCI -H460, Gl50 = 67.61±4.29 μg/ml. The lower activity against HepG2 cells might be related with the type of proliferation of this cell line, the only one with a monolayer growth type among the assayed lines . The activity demonstrated by the phenolic extract of C. monogyna might be explained by their high amounts in flavonoids, which are known for modulating a variety of biological events associated with cancer progression and development, such as cell proliferation, apoptosis, cell differentiation and neovascularization [3]. Accordingly, C. monogyna may be considered as a source of important phytochemicals with bioactive properties to be explored for pharmaceutical applications

Qualidade nutricional de agrião armazenado em atmosferas modificadas enriquecidas em gás inerte

O agrião (Nasturtium offidnale R. Br. ) é uma planta de crescimento rápido da família Brassicaceae consumida em sopas e saladas. Contudo, tal como a maioria dos vegetais, é altamente perecível. A utilização de embalagens com atmosfera modificada ou protetora tem aumentado nos últimos anos dado o seu potencial para prolongar a vida útil de vegetais minimamente processados. Esta técnica consiste em alterar a atmosfera dentro da embalagem, com o objetivo de abrandar o metabolismo e de prevenir reações de oxidação, evitando assim alterações de qualidade associadas a processos de maturação e senescência. A utilização de gases inertes tem sido testada, mas os estudos são ainda muito limitados. Neste sentido, este trabalho teve como objetivo avaliar os efeitos induzidos por diferentes atmosferas na qualidade nutricional de agrião minimamente processado durante o armazenamento refrigerado e selecionar o tratamento mais eficaz na extensão da vida útil. Amostras silvestres de agrião, recolhidas na região de Bragança, foram lavadas em água corrente e uma porção imediatamente analisada (controlo não armazenado). O restante material fresco foi embalado em atmosferas enriquecidas em azoto, árgon e ar (atmosfera controlo passiva), armazenado a 4°C durante 7 dias e posteriormente analisado. Determinou-se o teor em água, proteínas, lípides, glúcidos e cinzas e o valor energético seguindo métodos oficiais de análise; os perfis em açúcares, ácidos orgânicos, tocoferóis e ácidos gordos foram analisados por técnicas cromatográficas; e as propriedades antioxidantes avaliadas por ensaios químicos e bioquímicos. Tendo em conta a contribuição de todos os parâmetros de qualidade avaliados, foi possível concluir que o embatamento passivo foi o menos eficiente em preservar os atributos de qualidade característicos das amostras controlo não armazenadas. Por outro lado, o embalamento em atmosfera enriquecida em árgon foi a escolha mais acertada para preservar a qualidade global das amostras. Adicionalmente, este estudo destacou as propriedades nutricionais e antioxidantes do agrião, realçando assim o interesse da sua reinclusão nas dietas atuais

Electron beam and gamma irradiation as feasible conservation technologies for wild Arenaria montana L.: effects on chemical and antioxidant parameters

Wild plants are widely recognized as high-potential sources of several bioactive compounds. Nevertheless, these natural matrices require effective decontamination steps before they might be considered for different industrial purposes. Irradiation techniques are being progressively acknowledged as feasible conservation methodologies, either for their high decontamination effectiveness, as well as for their innocuousness on most chemical and bioactive parameters of the material to be treated. Arenaria montana L. (Caryophyllaceae) is recognized for its phytochemical richness, having a relevant geographical distribution in the Southern Europe. Herein the effects of irradiation (gamma and electron beam up to 10 kGy) were evaluated by comparing the nutritional, chemical and antioxidant profiles in A. montana extracts. In general, the assayed parameters showed statistically significant variations in response to irradiation treatment. Furthermore, the performed LDA allowed identifying the antioxidant indicators as the most affected parameters in irradiated samples, especially when using the 10 kGy dose and e-beam irradiation.Ministry of Agriculture, Portugal (Project PRODER/FEADER/EU no. 53514), for financial support of the work and E. Pereira grant; Foundation for Science and Technology (FCT, Portugal) for financial support to CIMO (PEst-OE/AGR/UI0690/2014), CTN (RECI/AAG-TEC/0400/2012), L. Barros (BPD/107855/2015) and J.C.M. Barreira (BPD/72802/2010). And to Prof A. Chmielewski, Director of INCT, and Dr A. Rafalski, for e-beam irradiations

Bryonia dioica, Tamus communis and Lonicera periclymenum fruits: characterization in phenolic compounds and incorporation of their extracts in hydrogel formulations for topical application

Plants are being increasingly used in dermatological formulations, since their collateral effects are lower than those caused by synthetic products. Hydrogels represent efficient formulations to incorporate plant extracts, providing good percutaneous absorption, non-greasy texture and easy application. Bryonia dioica Jacq. (white-bryony), Tamus communis L. (black-bryony) and Lonicera periclymenum L. (honeysuckle) fruits have important applications as topical homemade remedies. The phenolic extracts of B. dioica, T. communis and L. periclymenum were characterized by HPLC-DAD-ESI/MS, and further incorporated into hydrosoluble gels in order to evaluate their antioxidant potential. The hydrogels were prepared with each fruit extract at the EC50 value (obtained herein for each ethanolic extract). The antioxidant activity of extracts and hydrogels was evaluated following different chemical and biochemical assays. Eighteen compounds were identified in honeysuckle (7 flavonols, 7 hydroxycinnamoyl derivatives and 4 anthocyanins), fourteen in white-bryony (9 flavonols and 5 flavones) and twelve in black-bryony (7 flavonols and 5 flavones). Honeysuckle showed the highest concentrations of phenolic compounds, but the antioxidant activity was similar among the three assayed species. The obtained formulations revealed antioxidant activity close to 50% for reducing power, scavenging activity and β-carotene/linoleate assays. The studied fruits might be considered as good source of phenolic compounds that can be used in formulations for topical application, benefiting their antioxidant effects.Fundação para a Ciência e a Tecnologia (FCT, Portugal) for financial support to CIMO (strategic project PEst-OE/AGR/UI0690/2011). J.C.M. Barreira thanks to FCT, POPH-QREN and FSE for his grant (SFRH/BPD/72802/2010). M. Dueñas thanks to the Programa Ramón y Cajal for a contract. The GIP-USAL is financially supported by the Spanish Government through the Consolider-Ingenio 2010 Programme (FUN-C-FOOD, CSD2007-00063)

Targeting excessive free radicals with peels and juices of citrus fruits: grapefruit, lemon, lime and orange

A comparative study between the antioxidant properties of peel (flavedo and albedo) and juice of some commercially grown citrus fruit (Rutaceae), grapefruit (Citrus paradisi), lemon (Citrus limon), lime (Citrus aurantiifolia) and sweet orange (Citrus sinensis) was performed. Different in vitro assays were applied to the volatile and polar fractions of peels and to crude and polar fraction of juices: 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity, reducing power and inhibition of lipid peroxidation using b-carotene–linoleate model system in liposomes and thiobarbituric acid reactive substances (TBARS) assay in brain homogenates. Reducing sugars and phenolics were the main antioxidant compounds found in all the extracts. Peels polar fractions revealed the highest contents in phenolics, flavonoids, ascorbic acid, carotenoids and reducing sugars, which certainly contribute to the highest antioxidant potential found in these fractions. Peels volatile fractions were clearly separated using discriminant analysis, which is in agreement with their lowest antioxidant potential

Perennial spotted rockrose as a source of Ellagitannins with therapeutic interest: influence of drying and extraction conditions

The ellagitannins are a diverse class of hydrolysable tannins, a type of polyphenols present in some fruits, medicinal plants, nuts and seeds. They are complex derivatives of ellagic acid characterized by one or more hexahydroxydiphenoyl moieties esterified to a sugar, usually glucose [1]. These compounds play an important role in human nutrition and are endowed with numerous biological properties, including antioxidant, anti-inflammatory, anticancer, antiatherosclerotic, antihepatotoxic, antibacterial and anti-HIV replication activities [1-3], being therefore interesting molecules for medicinal and therapeutic applications. A previous work of our research team highlighted the perennial spotted rockrose (Tuberaria lignosa (Sweet) Samp.) as a rich source of ellagitannins, namely punicalagin [2]. In this study, the influence of drying and extraction conditions on the ellagitannins content from perennial spotted rockrose flowering aerial parts was evaluated. Wild specimens were gathered in the Northeast region of Portugal and then freeze-dried immediately after gathering or shade-dried in a dark and dry place at room temperature for 30 days. Subsequently, dried samples were extracted adding them to boiling water and left to stand at room temperature for 5 min, or boiling them for 5 min plus 5 min at room temperature. The ellagitannins analysis was performed by HPLC-DAD-ESI/MS. The effects of drying and extraction conditions were evaluated by analysis of variance (ANOVA) using the General Linear Model procedure. The most abundant ellagitannins were punicalagin isomers and punicalagin gallate isomers, being the highest levels detected in freeze-dried samples. Additionally, higher amounts of these compounds were achieved with longer extraction time (5 min + 5 min), in both shade- and freeze-dried samples. Thus, from the obtained results, it might be concluded that freeze-drying and longer extraction time are the preferable conditions to obtain high levels of ellagitannins with therapeutic interest from perennial spotted rockrose samples

Postharvest quality changes in fresh-cut watercress stored under conventional and inert gas-enriched modified atmosphere packaging

The effect of modified atmosphere packaging (MAP) on the postharvest quality of fresh-cut watercress (Nasturtium officinale R. Br.) stored at 4 ºC for 7 d was studied. A portion of watercress was immediately analyzed (non-stored control) and the remaining fresh material was stored packaged under atmospheres enriched with N2, Ar, air, or vacuum. The analyzed parameters included colour, total soluble solids, pH, macronutrients, the individual profiles of sugars, organic acids, tocopherols and fatty acids, and total phenolics and flavonoids. Furthermore, four in vitro assays were performed to evaluate the antioxidant activity. After assessing the effect on individual quality parameters, it was possible to conclude that air was the less efficient atmosphere in preserving quality attributes of the non-stored control samples during cold storage. In turn, Ar-enriched MAP was the most suitable choice to preserve the overall postharvest quality. The present study also highlighted the nutritional and antioxidant properties of watercress, as well as the interest of its inclusion in human diets.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support to CIMO (PEst-OE/AGR/UI0690/2014), REQUIMTE (UID/QUI/50006/2013), J. Pinela (SFRH/BD/92994/2013), J.C.M. Barreira (SFRH/BPD/72802/2010) and L. Barros (“Compromisso para a Ciência 2008” contract)

Development of paraben-free hydrogel based on plant extracts for topical application

Historically, medicinal preparations are derived from plants and their employment in dermatological and cosmetic products is increasing [1]. Topical application of products containing compounds with free radical scavenging properties protects tissues from oxidative damage [2]. The skin is an important protective barrier between the environment and the inner milieu, being highly exposed to oxidative stress, either from exogenous as well as endogenous sources [3]. The antioxidant potential of herbal extracts or pure isolated compounds have been extensively studied, but very few reports are available on the antioxidant properties of final formulations in which they have been included. In the present study, aqueous and ethanolic extracts from the flower buds of hawthorn (Crataegus monogyna Jacq.) were incorporated into hydrosoluble gels. Gels have been largely used in cosmetic products and as a dermatologic base, as they are easily dispersed, nonoily and can carry hydrosoluble active principles. Gel consistency was optimized when carbopol 940 was added at 1%. The prepared formulations presented a light green colour, a non-greasy texture and were promptly absorbed by the skin. Since the inclusion of parabens is nowadays poorly accepted by the consumers, imidazolidinyl urea was included as the antimicrobial component. Regarding pH evaluation, there were no significant alterations during the 90 days of observation, with values ranging between 5.5 and 6.5. The antioxidant activity of the prepared hydrogels was assessed and compared with a blank formulation (with all the components used in the hydrogel formulation, except the extract) and also with the results obtained for the extracts alone, at the same concentration (1 00 jlg/ml). The antioxidant activity measured in each hydrosoluble gel is very close to the value obtained for the isolated extract, in what regards inhibition of lipid peroxidation using thiobarbituric acid reactive substances (TBARS) and Beta carotene bleaching inhibition, 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity and reducing power. Hydrogels prepared with ethanolic extracts showed higher antioxidant activity than aqueous extracts, unless in Beta carotene bleaching inhibition assay. In general, it became evident that the inclusion of extracts in the prepared hydrosoluble gels caused very limited losses in their bioactivity