Flores das espécies Sambucus nigra, de origem européia, e Sambucus australis, nativa da América do Sul (Caprifoliaceae), denominadas sabugueiro e sabugueirodo- brasil, respectivamente, são utilizadas popularmente sob forma de infusão, como antiinflamatórias, laxativas e para condições febris resultantes de afecções do trato respiratório. Estudos prévios para S. nigra indicam compostos fenólicos como principais constituintes químicos, sendo estes relacionados às principais atividades biológicas avaliadas. Objetivando comparar esta espécie com Sambucus australis, foram realizadas análises botânicas macro e microscópicas das flores, identificando as principais diferenças entre as espécies, tais como o número de lóculos no ovário e presença de idioblastos cristalíferos em algumas estruturas, e observando os possíveis contaminantes (pedicelos). Também foram determinados os parâmetros farmacopéicos: cinzas totais e perda por dessecação. Após a análise química, foi escolhido o flavonóide rutina como marcador das espécies, para realizar análises quantitativas nas 31 amostras adquiridas e / ou coletadas, utilizando método de CLAE previamente validado. Soluções hidroetanólicas apresentaram maior capacidade de extração do produto alvo. Os limites mínimos de rutina observados para ambas as espécies foram de aproximadamente 0,65%. Também foram quantificados, por método espectrofotométrico, os flavonóides totais expressos em quercetina, sendo 0,93% e 1,46% os teores mínimos determinados para S.nigra e S. australis, respectivamente. O estudo da estabilidade acelerada (50°C ± 90% U.R.), avaliando a degradação dos constituintes químicos presentes permitiu sugerir a cinética de degradação de segunda ordem para da rutina nas duas espécies. Comparações de atividades biológicas das espécies foram realizadas pelos ensaios das atividades antiinflamatória (inibição do edema em pata de rato induzido por carragenina) e antioxidante (DPPH). Os resultados para o primeiro ensaio demonstraram ação equivalente em ambas as espécies para extratos hidroetanólicos a 80% (86% de inibição) e aquosos (81%), com atividade semelhante ao padrão indometacina (~83%); para a atividade antioxidante os extratos hidroetanólicos a 80% foram mais ativos (CE50 = 16 μg/ml) que os aquosos (CE50 = 27 μg/ml) em S. australis, e ambos extratos, superiores ao 28 extrato padronizado Gingko biloba (CE50 = 40 μg/ml) e aos extratos de S. nigra (CE50= 50 μg/ml – hidroetanólico e CE50= 32 μg/ml – aquoso). / Sambucus nigra (from Europe) and Sambucus australis (from South America) flowers (Caprifoliaceae), called sabugueiro and sabugueiro-do-brazil, respectively, are used in the folk medicine as anti-inflammatory, laxative and for respiratory diseases. Previous studies for S. nigra indicate phenolic compounds as the main chemical constituents, being related for these ones the biological activities reported. In order to compare this species with S. australis macro and microscopical analysis of the flowers were carried out to identify the major differences between the species, such as the number of ovary locules and idioblasts in some structures, verifying the possible contaminants (pedicels). The following Pharmacopoeia parameters were also determined: total ash and loss on drying. After the chemical analysis, it was chosen the rutin flavonol as the mark constituent for both species, to carry through quantitative analysis of the 31 acquired and/or collected samples, by previously validated HPLC method. Hydroethanolic solutions had shown greater capacity of extraction of the target product. The minimum limits of rutin observed for both species had been of approximately 0.65%. The total flavonoid content was also quantified by spectrophotometer method, being expressed as quercetin, finding 0.93% and 1.46% the minimum values for S. nigra and S. australis, respectively. The accelerated stability test (50 ºC ± 90% UR) that evaluate the degradation of the constituents, allowed to verify a kinetic degradation of second order for rutin in both species. Comparison of biological activities of these species was carried out using the antiedematogenic (rat paw edema) and antioxidant (DPPH) activities. The results for the first test showed an equivalent action for the hydroethanolic (86% of edema inibition) and aqueous (81%) extracts, exhibiting a similar activity to indometacin (~83%). For the antioxidant activity, the ethanolic extracts (CE50 = 16 μg/ml) were more active than the aqueous (CE50 = 27 μg/ml ) for S. australis, and, both extracts were superiors to the Ginko biloba (CE50 = 40 μg/ml) and to the S. nigra extracts (CE50 = 50 μg/ml – hydroethanolic and CE50 = 32 μg/ml - aqueous).

Orientador: Alba Regina Monteiro Souza Brito / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-18T00:28:36Z (GMT). No. of bitstreams: 1 Socca_EduardoAugustoRabelo_M.pdf: 1268769 bytes, checksum: eee2c6686e3a3fb123101e6954836a0f (MD5) Previous issue date: 2010 / Resumo: Retocolite ulcerativa idiopática e doença de Crohn são doenças inflamatórias intestinais caracterizadas por inflamação crônica da mucosa, resultando em diarréia, fezes sanguinolentas, dores abdominais, anemia, febre, fadiga e perda de peso, tanto em homens quanto em mulheres. Acredita-se que essas manifestações sejam resultado de uma interação multifatorial envolvendo indivíduos geneticamente susceptíveis, condições ambientais específicas, desbalanço na microflora intestinal e desajuste da resposta imune. Drogas derivadas do acido 5-aminossalicilico (sulfassalazina, mesalamina), corticosteroides e agentes imunomoduladores são utilizadas, em conjunto no tratamento dessas patologias. No entanto tais drogas apresentam efeitos adversos importantes, o que acaba por motivar pesquisas envolvendo produtos naturais como alternativas de tratamento. Neste contexto, este trabalho avaliou os efeitos de antocianinas presentes nos frutos de Sambucus nigra L. (sabugueiro), espécie arbustiva pertencente a família Adoxaceae, em modelo de colite ulcerativa induzida pelo acido 2,4,6-trinitrobenzeno sulfônico (TNBS). Os frutos do sabugueiro apresentam grandes concentrações de metabolitos secundários como antocianinas, alem de outros compostos fenólicos, que acabam por conferir aos frutos propriedades antioxidantes, anti-inflamatorias, imunomoduladoras e laxativas. Neste trabalho, foram avaliadas as propriedades antioxidantes e anti-inflamatorias da fração de antocianinas monoméricas presentes nos frutos de S. nigra. Os resultados comprovaram o efeito antioxidante in vitro das antocianinas, sendo elas eficazes em reduzir tanto o radical 2,2-difenil-1-picrilhidrazil (DPPH), teste que avalia a redução do radical via transferência de elétrons, quanto a taxa de oxidação do radical 2,2 azobis amidinopropano (AAPH), teste que verifica a oxidação do radical via transferência de átomos de hidrogênio. Nos ensaios in vivo concluiu-se que a dose de 5mg.Kg-1 de antocianinas apresentou a melhor resposta em reduzir a lesão causada pelo TNBS, sendo efetiva em manter os níveis de GSH (5,709 ± 0,931) comparado ao grupo TNBS (0,8525 ± 0,298) e ao grupo salina (6,610 ± 3,926). Essas antocianinas foram efetivas ainda em aumentar a atividade da SOD (8,487 ± 2,505), quando comparada com o grupo TNBS (3,884 ± 0,925) e ao grupo salina (12,240 ± 4,199), e reduzir a atividade da MPO (4,519 ± 2,016), quando comparada ao grupo TNBS (7,572 ± 2,572) e ao grupo salina (1,314 ± 0,319). Os resultados obtidos na avaliação da atividade das enzimas GPx (17,03 ± 3,951 no grupo tratado e 22,13 ± 11,510 no grupo TNBS) e GR (0,6524 ± 0,1180 no grupo tratado e 0,7249 ± 0,3968 no grupo TNBS), comparados ao grupo salina (84,22 ± 41,88) e (2,131 ± 0,9858) respectivamente, indicam que houve queda na ativação destas enzimas apos 24h de indução da colite, sendo que esta situação não foi revertida apos administração das antocianinas. Do mesmo modo não foram encontradas alterações nos níveis de LPO (5,756 ± 1,884 no grupo tratado e 5,113 ± 0,8254 no grupo TNBS) comparados ao grupo salina (4,688 ± 1,126) . Já os ensaios anti-inflamatórios revelaram atividade antiinflamatória promissora, visto que as antocianinas foram capazes de manter os níveis de IL-10 (466,7 ± 56,32) próximos aqueles obtidos no grupo não-colitico (492,4 ± 154,5), quando comparados ao grupo TNBS (264,8 ± 66,35), e de reduzir a produção de IL-12 (202,3 ± 53,33) quando comparados com os animais não tratados (319,3 ± 111,5) e ao grupo salina (149,8 ± 51,76). Neste estudo concluiu-se que as antocianinas presentes nos frutos de sabugueiro apresentam atividade antioxidante, alem de aumentar os níveis de IL-10, citocina essa que, provavelmente, participa na redução dos níveis de citocinas pro - inflamatórias como IL-12 e, consequentemente, a expressão de mediadores inflamatórios / Abstract: Ulcerative colitis and Crohn's disease are inflammatory bowel disease characterized by chronic inflammation of the mucosa, resulting in diarrhea, bloody stools, abdominal pain, anemia, fever, fatigue and weight loss in both men and women. It is believed that these manifestations are the result of a multifactorial interaction involving genetically susceptible individuals, environmental conditions, imbalance in intestinal microflora and immune response imbalance. Drugs derived from 5-aminosalicylic acid (sulfasalazine, mesalamine), corticosteroids and immunomodulatory agents are used together to treat these diseases. However, such drugs have significant adverse effects, which ultimately motivate research involving natural products as alternative treatments. In this context, this study evaluated the effects of anthocyanins in the fruits of Sambucus nigra L. (Elderberry), shrub species belonging to the family Adoxaceae, in a model of ulcerative colitis induced by 2,4,6-trinitrobenzenes sulfonic acid (TNBS). The fruits of elderberry have large concentrations of secondary metabolites such as anthocyanins and other phenolic compounds, which ultimately give the fruit antioxidant, anti-inflammatory, immunomodulatory and laxative properties. In this study we investigated the antioxidant and anti-inflammatory properties of the monomeric anthocyanins fraction in the fruits of S. nigra. The results confirmed the in vitro antioxidant effect of anthocyanins, which were effective in reducing both the 2,2-diphenyl-1-picrylhydrazyl (DPPH) test to evaluate the reduction of the radical via electron transfer and the rate of oxidation 2,2 azobis amidinopropane (AAPH), a test that checks radical oxidation via the transfer of hydrogen atoms. In vivo tests concluded that the dose of anthocyanins 5mg.Kg-1 had the best response to reduce the damage caused by TNBS, being effective in maintaining the levels of GSH (5.709 ± 0.931) compared to TNBS group (0, 8525 ± 0.298) and the saline group (6.610 ± 3.926). These anthocyanins were also effective in increasing the activity of SOD (8.487 ± 2.505) compared with the TNBS group (3.884 ± 0.925) and the saline group (12.240 ± 4.199), and reduce the activity of MPO (4.519 ± 2.016) when compared to TNBS group (7.572 ± 2.572) and the saline group (1.314 ± 0.319). The results obtained in the enzymatic activities of GPx (17.03 ± 3.951 in the treated group and 22.13 ± 11.510 in group TNBS) and GR (0.6524 ± 0.1180 in the treated group and 0.7249 ± 0.3968 TNBS group) compared to saline group (84.22 ± 41.88) and (2.131 ± 0.9858) respectively, indicate that there was a decrease in the activation of these enzymes after 24h of colitis induction. Likewise there were no changes in the levels of LPO (5.756 ± 1.884 in the treated group and 5.113 ± 0.8254 in group TNBS) compared to saline group (4.688 ± 1.126). The anti-inflammatory assays have shown promising antiinflammatory activity, whereas anthocyanins were able to maintain levels of IL-10 (466.7 ± 56.32) than those obtained in non-colitis group (492.4 ± 154 , 5), when compared to TNBS group (264.8 ± 66.35), and reduce the production of IL-12 (202.3 ± 53.33) compared with untreated animals (319.3 ± 111 , 5) and the saline group (149.8 ± 51.76). In this study it was concluded that anthocyanins present in elderberry fruits have antioxidant activity, and increased levels of IL-10, this cytokine, which probably participates in reducing levels of proinflammatory cytokines such as IL-12 and consequently the expression of inflammatory mediators / Mestrado / Fisiologia / Mestre em Biologia Funcional e Molecular

This diploma thesis is concerned with determination anthocyanins dyes in elderberry (Sambucus nigra L.). In theoretical part there is described general characterization of the plant, its chemical composition, short view of elderberry breeding and possibilities of usage in food and medical industry. Bigger attention is paid to anthocyanins dyes. There is described their characterization, occurrence, factors that have an impact on their stability. Biosynhesis of the pigments and metabolism, benefit effects and their usage in food chemistry are then shortly sumarized. Methods of determination of anthocyanins are discused too. Next chapter deals with high performance liquid chromatography. At the end of theoretical part, there is sumarized findings of anthocyan dyes analysis by expert scientists in various matrix. In experimental part, total of 16 species of elderberry were analyzed. These species are from research and breeding institute of pomology in Holovousy. The dyes were identification and quantification by high performace liquid chromatography coupled with a diode array detector. Detection wavelenght was 520 nm. Gradient of mobile phase was optimilized because of shortening analyses time and resolution improvement. Four dyes were identified by HPLC: cyanidin-3-sambubioside, cyanidin-3-glukoside, cyanidin-3,5-diglukoside a cyanidin-3-sambubioside-5-glukoside. Total amount of anthocyanins ranged from 33627167,0 to 5525284,0 mAU•min-1. The most anthocyanins dyes were determined in Mammut species (33627167,0 mAU•min-1), the lowest amount was measured in Aurea species (5525284,0 mAU•min-1). The most abundant anthocyanin in all species of eldeberry fruit except Haschebrg and Aurea species was cyanidin-3-sambubioside. The concentrations of cynidin-3-glucoside in all of the berries were quantified using regression equation of calibration curve. Mammut species had the most amount of this anthocyan (0,486 mg•ml-1) and the lowest amount had Aurea species (0,033 mg•ml-1).

This work deals with the elemental analysis of different elderberry (Sambucus nigra L.) cultivars. In the theoretical part the botanical characteristics of elderberry, including its extension and places of occurrence, the possibilities of collecting the various parts of the elderberry and its use in food and cosmetics is described. A large part is devoted to the occurrence and characteristics of minerals in food, and to the methods that are suitable for elemental analysis and decomposition of biological material. The aim of the experimental part was to analyze different cultivars of elderberry and juice made from this cultivars, and to determine concentration of selected elements (Zn, Mn, Fe, Mg, Cu, P, Ca, Na and K) using ICP–OES. The total amount of 24 cultivars was analysed from which 6 varieties were harvested in two different seasons in 2011 and 2013. Measured concentrations of elements in different varieties of elderberry were mutually compared using statistical methods. The differences between the content of elements in fruits and juice, obtained by pressing the fruit, and the effect of seasonal conditions on the content of elements in the fruit juice from different elderberry cultivars were studied.

Amount of rutin and quercetin in bark and leaves from different type of Sambucus nigra was determined in this diploma thesis. Records of amounts were compared with amounts in significant sources of these flavonoids. Matters from raw matrix were obtained with pressurized solvent extraction (PSE) and pressurized hot water extraction (PHWE). There was use temperature 120°C, pressure 150 bar, three times 5-min cycles for PSE. There were used 80°C, 150 bar, three 5-min cycles for rutin extracted with PHWE and the same condition were used for extraction of quercetin only the temperature was changed to 100 °C. Analysis of rutin and quercetin were carry out high performance liquid chromatography (HPLC) with diode array detection (DAD), where was used SUPELCOSILTM LC 8DB column (5µm; 250 x 4.6 mm). Methanol: water: formic acid (36: 61.5: 2.5, pH 2.17 2.28) was used like a mobile phase. The flow of the mobile phase was set up 0.7 ml/min. Size of sample was 10 µl. The most amount of rutin was determined in the leaves of wild kind of Sambucus nigra with methanol extraction and hot water too. Extraction with PHWE was more effectively. There were determined 5.58 mg of rutin in one gram of sample from wild Sambucus nigra. There were determined the smallest amount of rutin in Sambucus nigra Körsör. There was found only 0.13 mg/g with extraction PHWE. The quercetin wasn’t determined in any Sambucus nigra leaves. There were found both of the flavonoids (rutin and quercetin) in the branch of Sambucus nigra. The rutin was determined in the small amount in wild kind but it was found in the important amount in Sambucus nigra cultivar. Whereas the quercetin was found in higher concentration than rutin when was finding the ideal conditions for extraction of branch. The quercetin was chosen like more important in light of amount. In the end the quercetin wasn’t detected in any Sambucus nigra cultivars. The most amount of quercetin was determined in wild sambucus nigra, 0.24 mg/g. The most amount of rutin was found in Sambucus nigra Albida, 2.33 mg/g.

My diploma thesis deals with the optimization and verification of conditions for separation of proteins, which are contained in the twigs of elder (Sambucus nigra L.) using the technique RP-HPLC. The measurements were made with the use of a HPLC system with a UV-VIS detector. Column Zorbax 300SB-C18 300 4,6 x 250 mm and particles the size of 5 microns were used. The theoretical part describes the attributes and usage of elder and especially its proteins. The basic characteristic of the used HPLC technique is introduced. The possibilities of how to identify proteins are described. The practical part demonstrates the individual steps of optimization of the HPLC method applied to the mixture of standard proteins. The application of this method in the real sample is also reported. The final part of the diploma thesis is focused on the comparison of analysed samples taken in different seasons.

An evidence-based systematic review of elderberry and elderflower (Sambucus nigra) by the Natural Standard Research Collaboration consolidates the safety and efficacy data available in the scientific literature using a validated, reproducible grading rationale. This article includes written and statistical analysis of clinical trials, plus a compilation of expert opinion, folkloric precedent, history, pharmacology, kinetics/dynamics, interactions, adverse effects, toxicology, and dosing.

In this thesis was determinated selected chemical and nutritional parameters in the wild elderberry and 17 cultivated varieties of elderberry. The fruits was provided by the Research and Breeding Institute of Pomology Holovousy Ltd. The theoretical part deals describes elderberry, chemical composition of elderberry, especially biological active substances and its using not only food industry. Closer attention has been focused on effect of saccharides, anthocyanins and methods of their determination, specially determination by HPLC. The experimental part describes the various methods of determination of selected chemical and nutritional parameters. Total dry matter, content of total polyphenolic compounds, monomeric anthocyanin pigment and antioxidant activity was defined in all varieties of elderberry fruits. For some varieties was defined total soluble dry matter, content of organic acids and saccharides by various methods. In the end was compared the values from different methods varieties.

Made available in DSpace on 2014-12-17T14:13:40Z (GMT). No. of bitstreams: 1 CamilaGR.pdf: 488995 bytes, checksum: fa9e656a33d6c75d3226bec90875615a (MD5) Previous issue date: 2008-08-12 / Radiobiocomplexes are used in nuclear medicine to obtain images and to treat diseases. Blood constituents have been used as radiobiocomplexes. Natural or synthetic products can influence on the labeling of blood constituents with technetium-99m (99mTc), the morphology of red blood cells and on the stannous chloride (SnCl2) action on plasmid DNA. Sambucus australis and Sambucus nigra are used in popular culture for treating diseases. The aim of this work was to evaluate the effects of the extracts of Sambucus australis and Sambucus nigra on the labeling of blood constituents with 99mTc, on morphology of red blood cells of Wistar rats, on the topology of plasmids DNA and the action against the SnCl2 effects on the DNA of plasmids pBSK. On the labeling of blood constituents with 99mTc it was verified that both extracts were capable to decrease significantly the radioactivity in the cellular compartment and in the insoluble fraction of plasma. Sambucus australis also decreased the labeling of insoluble fraction of blood cells with 99mTc. Both extracts did not alter the morphology of red blood cells. Moreover, it was verified that Sambucus nigra did not alter the electrophoretic profile of plasmid DNA, but decreased the effect of SnCl2 on plasmid DNA. These last results sugest a genotoxic effect and a protective action of Sambucus nigra extract against the SnCl2 action on plasmid DNA. This work was developed with the contribution of several Departments of biomedical area of the Hospital Universit?rio Pedro Ernesto, of the UERJ, characterizing a multidisciplinary experimental research / Radiobiocomplexos s?o utilizados em medicina nuclear para obten??o de imagens e tratamento de doen?as. Constituintes sangu?neos t?m sido utilizados como radiobiocomplexos. Produtos naturais ou sint?ticos podem influenciar a marca??o de constituintes sang??neos com tecn?cio-99m (99mTc), a morfologia de hem?cias e a a??o do cloreto estanoso no DNA plasmidial. Sambucus australis e Sambucus nigra s?o utilizados na cultura popular para o tratamento de doen?as. O objetivo desse estudo foi avaliar os efeitos dos extratos de Sambucus australis e de Sambucus nigra na marca??o de constituintes sang??neos com 99mTc, na morfologia de hem?cias de ratos Wistar, na topologia do DNA plasmidial e sua a??o sobre os efeitos do cloreto estanoso no DNA de plasm?dios pBSK. Na marca??o de constituintes sang??neos com 99mTc verificou-se que os dois extratos foram capazes de diminuir significativamente a radioatividade no compartimento celular e na fra??o insol?vel do plasma. Sambucus australis diminuiu tamb?m a marca??o da fra??o insol?vel da c?lula com 99mTc. Os dois extratos n?o alteraram a morfologia das hem?cias. Al?m disso, foi verificado que um extrato de Sambucus nigra n?o alterou o perfil eletrofor?tico do DNA plasmidial, entretanto diminuiu os efeitos do cloreto estanoso no DNA plasmidial. Estes ?ltimos resultados sugerem um efeito genot?xico e uma a??o protetora do extrato de Sambucus nigra contra a a??o do cloreto estanoso no DNA plasmidial. Este trabalho foi realizado com a colabora??o de diversos Departamentos da ?rea biom?dica do Hospital Universit?rio Pedro Ernesto, da UERJ, caracterizando uma pesquisa experimental multidisciplinar

This Diploma thesis deals with the production of the concentrate from the Elderberry marc extract (Sambucus nigra L.). The basic chemical characteristics in this concentrate were determined. The theoretical part of the thesis deal with the botanic characteristic of the Elderberry and its berries. The characteristic of the phenolic compounds, mainly anhocyanins was summarized. The use of the anthocyanin pigments in the food industry, the possibilities of their extraction and the concentrate production was discussed. Thereafter the methods of the anthocyanin content and the phenolic content determination were described. In the experimental part of study, the production of the anthocyanin rich extract from Elderberry marc was optimized. The extraction of the marc by the 1:1 ethanol/water (V/V) solvent with the ratio marc/solvent 1:2 (w/V) showed the most convenient. The production of the concentrate followed. During the concentration on the rotary evaporator the pigment degradation was investigated. The degradation decreased with the decrease of the temperature. The final concentrate was prepared from the extract on the rotary evaporator by 45 °C for 15 minutes. This product had the 7times lower volume compared to the initial extract. In the final concentrate come of the chemical characteristic were determined. By liquid chromatography (HPLC) the major pigments cyanidin-3-sambubiosid and cyanidin-3-glucosid were determined. Their total content was determined to 34,9 gl-1. The total anthocyanin content was determined by pH differential method to 7,1 gl-1 (expressed as cyanidin-3-glucoside equivalent). The total phenolic content was determined to 27,1 gl-1 (expressed as gallic acid equivalent) by Folin-Ciocalteu method. By electron paramagnetic resonance the increase in antioxidant activity with the increase in concentration was found. Up to 42 aromatic compounds was determined in Elderberry marc, its extract and the final concentrate.