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- Componente volátil de lúpulo: uma análise comparativa entre clones espontâneos e variedadesPublication . Goes, Hugo; Pedro, Luís G.; Sousa, Maria JoãoO Humulus lupulus L. é uma espécie pertencente à família Cannabaceae. O lúpulo, como é vulgarmente conhecido, é uma planta herbácea perene, dióica e normalmente diplóide (2n=20) [1]. É na produção de cerveja que o lúpulo apresenta o seu maior valor económico a nível internacional. Foi na Flandres que começou verdadeiramente a produção de cerveja, uma vez que desde o início da sua fabricação na Mesopotâmia, até então, a cerveja era uma bebida adocicada, baseada na fermentação de malte, com o nome de Ale sendo a sua conservação um dos maiores problemas. A utilização de lúpulo veio resolver esse problema, visto que produz compostos com acção bactericida, em particular contra bactérias Gram-negativas [2]. A sua produção estendeu-se por toda a Europa e, daí, ao resto do mundo. Actualmente a procura de novos aromas tem vindo a aumentar, impulsionada pela recente expansão da produção artesanal de cerveja em Portugal. Uma vez que existe lúpulo espontâneo em grande parte do país, a recolha e análise dos aromas desses lúpulos poderá levar-nos ao desenvolvimento de novos e mais aromas. As componentes voláteis extraídas de lúpulos espontâneos recolhidos na zona de Bragança, foram analisadas e comparadas com as de variedades comerciais. Material e métodos: O material vegetal foi colhido em diferentes zonas do distrito de Bragança, junto a cursos de água. Os cones femininos foram recolhidos e os voláteis foram extraídos num sistema Likens-Nickerson e posteriormente analisados por CG e CG-EM. Resultados e conclusão: Os resultados obtidos evidenciam uma certa semelhança na componente monoterpénica, com o β-mirceno como composto maioritário (75 e 64%, respectivamente na amostra do cultivar e na amostra do espontâneo) e diferenças notórias na componente sesquiterpénica, bem evidente nos casos do α-humulene (12% no cultivar, 0,2% no espontâneo) e do trans-β-farnesene (não detectado no cultivar, 9% no espontâneo). É também merecedor de realce a maior riqueza da componente sesquiterpénica do clone espontâneo, em particular nos compostos oxigenados
- Obtenção de plantas de Humulus lupulos L. resistentes a virusPublication . Sousa, Maria JoãoA biotecnologia vegetal tornou-se uma ferramenta fundamental no melhoramento das mais variadas espécies. Na sua vertente de transformação genética, abriu novos horizontes de interesse agrícola e económico, no controlo de doenças e pragas. No lúpulo, a perda de produtividade, devido a infeções virais, levou a que a estratégia de transformação genética, para introdução de resistência a vírus, se tornasse extremamente interessante. Neste trabalho, procurou-se estabelecer condições para obter um sistema de transformação eficiente, com base na introdução do gene da cápside viral. Testou-se o sistema de micropropagação e de regeneração em duas cultivares (Eroica e Brewer’s Gold) e num clone espontâneo (Bragança). Utilizaram-se diferentes meios base e diferentes explantes (folhas, pecíolos e entrenós). Este sistema de regeneração, permitiu desenvolver dois processos de transformação: 1) mediado por Agrobacterium e 2) bombardeamento de partículas. Foram testados diferentes parâmetros de transformação (plasmídeos, tempos de corte, co-cultura, meios de seleção). Verificou-se a transformação por diferentes técnicas de análise molecular e obteve-se transformação estável em qualquer dos processos. Após 2 anos em terra, as plantas transformadas utilizando os dois processos, foram testadas para o gene da cpArMV, apresentando resultados positivos.
- Humulus lupulus L. analysis and comparison of volatile of spontaneous and commercial varieties: phytotherapeutic potentialitiesPublication . Goes, Hugo; Sousa, Maria João; Pedro, Luís G.Humulus lupulus L. is a species in the Cannabaceae family. Hop, as it is commonly known, is a perennial, dioecious and normally diploid (2n = 20) herbaceous plant [1]. It is in beer production that hops have their greatest economic value at the international level. Due to the production of compounds with bactericidal action, in particular against Gramnegative bacteria, the hops came to solve problems related to the conservation of beer [2]. In addition Hops contain compounds that confer sedative, diuretic and antiarthritic properties [3]. In the ethnobotanical use, the dried flowers were used in pillows called "hop pads", to combat insomnia [4]. The use of dried and green hop inflorescences for diuretic uses and disorders of the digestive tract, were verified in the Montesinho park area [5]. In China, alcoholic extracts of hops were used to treat leprosy, tuberculosis and dysentery [2]. Its use in infusions as a tonic, since the middle Ages where it considered a medicinal herb [6]. The results obtained show a certain similarity in the monoterpene component, with β-myrcene as the major compound (more than 64% in cultivar and spontaneous samples) and notorious differences in the sesquiterpene component, evident in cases of α-humulene (12% in cultivars, 0.2% and in spontaneous) and trans-β-farnesene (not detected in cultivars, 9% in spontaneous). Also important is the richness of the sesquiterpene component in spontaneous clone, in particular in the oxygenated compounds. This profile, of both cultivar and spontaneous, shows potential for antiseptic effects in the digestive tract, especially when these are associated with bacteria such as Helicobacter pylori [7], which will be in agreement with ethnobotanical uses of Hop. Since there is spontaneous hops in a large part of Portugal, the collection and analysis of the aromas of these hops may lead to the development of new and more fragrances, with interest in different areas, like beer production or cosmetics. The volatile components extracted from spontaneous hops collected in the Bragança area were analyzed and compared with those of commercial varieties.
- Development of a cosmetic anti-aging gel formulation with extracts of spontaneous plants and cultivars of Humulus lupulus L.Publication . Santos, Briolanja; Pereira, Olívia R.; Afonso, Andrea Luísa Fernandes; Pedro, Luís G.; Cardoso, Susana M.; Sousa, Maria JoãoHop (Humulus lupulus L.) is known worldwide as an essential flavor in the beer industry. Its major compounds have been demonstrated to be associated to health benefits, due to its antimicrobial, antioxidant, anti-inflammatory and anticancer activities [1]. This study intended to develop and to evaluate the stability of an anti-aging cosmetic gel by incorporation of different percentages of hydroalcoholic extracts obtained from cones (flowers) and vegetative parts (leaves) of hop spontaneous and of the Cascade and Polaris cultivars and essential oil of Thymus zygis. In addition, the phytochemical profile and antimicrobial activities determination of hop extracts and of essential oil of thymus were performed. A methylcellulose gel formulation was used as base to develop the hop cosmetic product. Subsequently, several parameters were evaluated to address its physical-chemical stability. In addition to that, the phenolic profile of hop extracts was determined by UHPLC-DAD-ESI-MS2 [2] while the composition of thyme essential oil was done by GC and GC-MS. The ability to inhibit the microbial growth was performed by microdilution broth method and the diffusion in agar test. Contrary to hop extracts, thyme oil has shown to exert antibacterial and antifungal activities and thus, it was added to the formulation as preservative. Carvacrol, terpinen-4-ol and p-cimene, determined as major compounds in this oil, could be important contributors for these activities. The extracts obtained from hop cones were rich in phenolic compounds such as cohumulone, humulone and xanthohumol, which have been claimed as possessing anti-wrinkle effect [3]. Concerning stability of the formulations, no phase separation was observed in the centrifugation, mechanical vibration tests or the color tests. However, changes were observed in the UV-Visible spectra between 300nm to 350nm wavelengths, probably by presence of the phenolic compounds such as example xanthohumol, cohumulona, humulona and lupulona [3]. In texture by back extrusion, only the formulation with the extracts of the Polaris variety has high consistency and low viscosity. In the light test there was phase separation in the samples due to the occurrence of dehydration, to minimize this effect, it must be properly in a non-transparent packaging to protect from light. Overall, the incorporation of hydroalcoholic extracts of hop and essential oil of thyme in methylcellulose gel formulations, especially with Polaris variety with 2.5% hydroalcoholic extract of hop cones, allow to obtained a stable cosmetic with potential as anti-aging take into account the chemical composition.
- Hop by-products: pharmacological activities and potential application as cosmeticsPublication . Pereira, Olívia R.; Santos, Gleiciara; Sousa, Maria JoãoHops (Humulus lupulus L.) are known worldwide as a raw material in beer production due their flavor and preservative values. The beneficial properties of the plant have been mostly associated with the female hop inflorescences (cones), which is also the part used in the brewing industry. However, some studies indicate the presence of compounds associated with health benefits in the vegetative parts of hops or small-caliber cones, which discarded in hop collection. Moreover, large quantities of by-products remain in the forms of spent grains and spent hops/hot trub and are produced by breweries raising environmental and economic sustainability concerns. This review focuses on the phytochemicals and biological and pharmacological activities of hop and their potential use in skin care products and also intends to explore the potential of the hop’ discarded parts and brewery industry by-products for production in the cosmetics industry.
- Humulus lupulus L.: cosmetic application of extracts obtained from cones and vegetative partsPublication . Alves, Silena Silva Delgado; Afonso, Andrea Luísa Fernandes; Cardoso, Susana M.; Sousa, Maria João; Pereira, Olívia R.Nowadays there is an increasing interest in the search for bioactive compounds obtained from aromatic and medicinal plants. Hop (Humulus lupulus L.) is known worldwide as an essential raw material for the beer industry, however, recent studies have revealed health benefits. In the beer production process, only the cones of hop are used, while the vegetative part of the plant is discarded and not valorized albeit its richness in interesting compounds. In this work, we have studied the phytochemical profile of polar extracts of hop and of the essential oil of T. zygis subsp. zygis, and further evaluated their antibacterial and antifungal capacities. Moreover, gel formulations incorporating distinct levels of hop extracts (1.25%, 2.5% and 5%) from cones and vegetative parts of the varieties Nugget and Polaris of hop and spontaneous hop, combined with essential oil of Thymus zygis subsp. zygis, were developed and the stability of the new cosmetic products was evaluated. UHPLC-DAD-ESI-MS2 analysis was used for phytochemical study of hydroalcoholic hop extracts. The antimicrobial properties of these extracts and of the essential oil of T. zygis subsp. zygis were determined by microdilution broth method and the diffusion in agar test. Additionally, the stability of the gel formulations was evaluated through a series of physical-chemical tests such as the pH, density, texture evaluation, mechanical vibration, centrifugation, relative humidity, light test, consecutive extreme temperature variation and evaluation of organoleptic characteristics. The stability tests did not reveal phase separation or liquefaction of the prepared gel samples. There were no changes recorded in vibration test, in the organoleptic characteristics of the formulations, the relative humidity test, as well as in the extreme temperature variation test. The formulations showed an acid pH, between 5.93±0.047 and 6.16±0.047. The apparent density of the formulations were between 1.08±0.00 Kg/m-3 and 1.12±0.00 Kg/m-3. Although hop extracts had no antimicrobial effect, the essential oil of T. zygis subsp. zygis showed antibacterial and antifungal activities against all strains of bacteria and fungi tested, with higher potency against S. aureus (MIC = 0.031%) than for P. aeruginosa (MIC = 1.25%). The study of the phenolic profile of cones and vegetative parts of the polar extracts of hop, allowed the identification of thirty compounds, of which alpha and beta acids, phenolic acids and flavonoids. In conclusion, the study explored the vegetative part of hop that remain poorly studied allowing propose them as ingredient in cosmetic area.
- Development of a cosmetic anti-aging gel formulation with extracts of spontaneous plants and cultivars of Humulus lupulus L.Publication . Santos, Briolanja; Pereira, Olívia R.; Afonso, Andrea Luísa Fernandes; Pedro, Luís G.; Cardoso, Susana M.; Sousa, Maria JoãoHop (Humulus lupulus L.) is known worldwide as an essential flavor in the beer industry. Its major compounds have been demonstrated to be associated to health benefits, due to its antimicrobial, antioxidant, anti-inflammatory and anticancer activities [1]. This study intended to develop and to evaluate the stability of an anti-aging cosmetic gel by incorporation of different percentages of hydroalcoholic extracts obtained from cones (flowers) and vegetative parts (leaves) of hop spontaneous and of the Cascade and Polaris cultivars and essential oil of Thymus zygis. In addition, the phytochemical profile and antimicrobial activities determination of hop extracts and of essential oil of thymus were performed. A methylcellulose gel formulation was used as base to develop the hop cosmetic product. Subsequently, several parameters were evaluated to address its physical-chemical stability. In addition to that, the phenolic profile of hop extracts was determined by UHPLC-DAD-ESI-MS2 [2] while the composition of thyme essential oil was done by GC and GC-MS. The ability to inhibit the microbial growth was performed by microdilution broth method and the diffusion in agar test. Contrary to hop extracts, thyme oil has shown to exert antibacterial and antifungal activities and thus, it was added to the formulation as preservative. Carvacrol, terpinen-4-ol and p-cimene, determined as major compounds in this oil, could be important contributors for these activities. The extracts obtained from hop cones were rich in phenolic compounds such as cohumulone, humulone and xanthohumol, which have been claimed as possessing anti-wrinkle effect [3]. Concerning stability of the formulations, no phase separation was observed in the centrifugation, mechanical vibration tests or the color tests. However, changes were observed in the UV-Visible spectra between 300nm to 350nm wavelengths, probably by presence of the phenolic compounds such as example xanthohumol, cohumulona, humulona and lupulona [3]. In texture by back extrusion, only the formulation with the extracts of the Polaris variety has high consistency and low viscosity. In the light test there was phase separation in the samples due to the occurrence of dehydration, to minimize this effect, it must be properly in a non-transparent packaging to protect from light. Overall, the incorporation of hydroalcoholic extracts of hop and essential oil of thyme in methylcellulose gel formulations, especially with Polaris variety with 2.5% hydroalcoholic extract of hop cones, allow to obtained a stable cosmetic with potential as anti-aging take into account the chemical composition.
- Humulus lupulus L. analysis and comparison of volatile of spontaneous and commercial varieties: phytotherapeutic potentialitiesPublication . Goes, Hugo; Sousa, Maria João; Pedro, Luís G.Humulus lupulus L. is a species in the Cannabaceae family. Hop, as it is commonly known, is a perennial, dioecious and normally diploid (2n = 20) herbaceous plant [1]. It is in beer production that hops have their greatest economic value at the international level. Due to the production of compounds with bactericidal action, in particular against Gramnegative bacteria, the hops came to solve problems related to the conservation of beer [2]. In addition Hops contain compounds that confer sedative, diuretic and antiarthritic properties [3]. In the ethnobotanical use, the dried flowers were used in pillows called "hop pads", to combat insomnia [4]. The use of dried and green hop inflorescences for diuretic uses and disorders of the digestive tract, were verified in the Montesinho park area [5]. In China, alcoholic extracts of hops were used to treat leprosy, tuberculosis and dysentery [2]. Its use in infusions as a tonic, since the middle Ages where it considered a medicinal herb [6]. The results obtained show a certain similarity in the monoterpene component, with β-myrcene as the major compound (more than 64% in cultivar and spontaneous samples) and notorious differences in the sesquiterpene component, evident in cases of α-humulene (12% in cultivars, 0.2% and in spontaneous) and trans-β-farnesene (not detected in cultivars, 9% in spontaneous). Also important is the richness of the sesquiterpene component in spontaneous clone, in particular in the oxygenated compounds. This profile, of both cultivar and spontaneous, shows potential for antiseptic effects in the digestive tract, especially when these are associated with bacteria such as Helicobacter pylori [7], which will be in agreement with ethnobotanical uses of Hop. Since there is spontaneous hops in a large part of Portugal, the collection and analysis of the aromas of these hops may lead to the development of new and more fragrances, with interest in different areas, like beer production or cosmetics. The volatile components extracted from spontaneous hops collected in the Bragança area were analyzed and compared with those of commercial varieties.