Identificação de cromenos, naftoquinonas e outros constituintes de Sinningia allagophylla (Gesneriaceae) e avaliação de suas atividades citotóxica, anti-inflamatória, analgésica e larvicida

Orientador : Prof. Dr. Maria Élida Alves StefanelloTese (doutorado) - Universidade Federal do Paraná, Setor de Ciências Exatas, Programa de Pós-Graduação em Química. Defesa: Curitiba, 24/03/2014Inclui referênciasResumo: A familia Gesneriaceae compreende cerca de 150 generos e 3000 especies, distribuidas nas regioes tropicais e subtropicais do mundo inteiro. No Brasil esta representada por 28 generos e 215 especies. O genero Sinningia compreende 68 especies, das quais 67 sao encontradas em territorio brasileiro. Os estudos quimicos ja realizados revelam que as especies de Sinningia contem principalmente antraquinonas e terpenos. Sinningia allagophylla (Martius) Wiehler, e uma erva anual, com tuberculo perene, encontrada nas regioes Centro-Oeste, Sul e Sudeste do Brasil, e tambem na Argentina e no Paraguai. E considerada medicinal, sendo usada na medicina popular como tonico, febrifugo, depurativo e diuretico. O presente trabalho descreve, pela primeira vez, o isolamento e identificacao de seus constituintes quimicos e a avaliacao das atividades citotoxica, analgesica, anti-inflamatoria e larvicida de extratos e substancias isoladas. Os tuberculos secos de S. allagophylla foram extraidos com etanol. O extrato etanolico foi fracionado por particao com solventes e, posteriormente por cromatografia. As substancias puras obtidas foram identificadas por tecnicas espectrometricas (massas, RMN). Desse modo foram identificadas 23 substancias: lapachenol (1), 8-metoxilapachenol (2), 8- metoxi-12-hidroxilapachenol (3), allagophyllolideo (4), allagophyllina A (5), allagophyllina B (6), allagophyllina C (7), tectoquinona (8), dunniol (9), -dunniona (10), 8-hidroxidunniona (11), dunniona (12), allagophyllona (13), aggregatina E (14), 6-metoxiaggregatina E (15) halleridona (16), cedrol (17), -muurolol (18), 8-cedren-13-ol (19), oleato de sitosterila (20), linoleato de sitosterila (21), oleato de estigmasterila (22) e linoleato de estigmasterila (23). As substancias 2-7 e 13-15 sao ineditas. O extrato etanolico apresentou atividade citotoxica, analgesica e anti-inflamatoria. A atividade citotoxica foi avaliada in vitro contra varias linhagens de celulas tumorais humanas, sendo 9 identificada como a substancia mais ativa. Esta naftoquinona inibiu totalmente o crescimento das linhagens de celulas tumorais de leucemia (K562), glioma (U251), mama (MCF-7), ovario (OVCAR-3) e ovario resistente a multiplos farmacos (NCI-ADR/RES) em concentracoes de 1,14-6,88 ƒÊg mL-1 (4,5-28 ƒÊM). As atividades analgesica e anti-inflamatoria foram avaliadas in vivo, em camundongos. A substancia 2 foi ativa na concentracao de 1,8 mg kg-1 (6,7 ƒÊmol kg-1), quando administrada por via oral. Essa substancia tambem apresentou atividade larvicida contra larvas de Aedes aegypti, com um valor de CL50 = 7,41 ƒÊg mL-1 (27,4 ƒÊM) apos 24 h.Abstract: The family Gesneriaceae comprises about 150 genus and 3000 species, distributed in the tropics and subtropics around the world. In Brazil the family is represented by 28 genera and 215 species. The genus Sinningia comprises 68 species, of which 67 are found in Brazil. Previous chemical studies reveal that Sinningia species produce mainly anthraquinones and terpenes. Sinningia allagophylla (Martius) Wiehler is an annual herb, with perennial tubers, growing in the Mid-Western, Southeastern and Southern regions of Brazil, besides Argentine and Paraguay. This plant is considered medicinal, being used in the folk medicine as tonic, febrifuge, depurative and diuretic. This work describes, for the first time, the isolation and identification of chemical constituents and evaluation of cytotoxic, analgesic, antiinflammatory and larvicidal activities of extracts and pure compounds of S. allagophylla. Dried tubers were extracted with ethanol. The extract was fractionated by partition with solvents and chromatographic techniques. Pure compounds were identified by spectrometric techniques (mass, NMR). This procedure resulted in the identification of 23 compounds: lapachenol (1), 8-methoxylapachenol (2), 8-methoxy-12-hydroxylapachenol (3), allagophyllolide (4), allagophyllin A (5), allagophyllin B (6), allagophyllin C (7), tectoquinone (8), dunniol (9), -dunnione (10), 8-hydroxydunnione (11), dunnione (12), allagophyllone (13), aggregatin E (14), 6-methoxyaggregatin E (15), halleridone (16), cedrol (17), -muurolol (18), 8-cedren-13-ol (19), sitosteryl oleate (20), sitosteryl linoleate (21), stigmasteryl oleate (22) and stigmasteryl linoleate (23). Compounds 2-7 and 13-15 are new. The ethanol extract showed cytotoxic, analgesic and anti-inflammatory activities. The cytotoxic activity was evaluated in vitro against several human tumor cell lines, and 9 was identified as the most active compound. This naphtoquinone inhibited totally the growth of leukemia (K562), gliome (U251), mama (MCF-7), ovarian (OVCAR-3), and drug-resistant ovarian cell lines with concentrations of 1.14-6.88 ìg mL-1 (4.5-28 ìM). The analgesic and anti-inflammatory activities were evaluated in vivo, using mice. Compound 2 showed activity in the concentration of 1.8 mg kg-1 (6.7 ìmol kg-1), by via oral. This compound also exhibited larvicidal activity against Aedes aegypti larvae, with LC50 = 7.41 ìg mL-1 (27.4 ìM) after 24 h

Biogas production and electricity generation from a quail manure wastewater treatment system per water depth

Coturniculture, as an activity which demands low investment and quick return, is shown to be a possibility for the rural family producer. Concomitant to this, we highlight the fact that the use of liquid quail farming waste, aimed at generating energy by anaerobic digestion, can mean a viable and promising technology for obtaining biogas from confined animal production systems. The growing demand for energy establishes that new energy sources are better used, and a great opportunity for their growth may be the use of biomass in anaerobic digestion systems, in which the organic substrate is degraded and transformed into energy and biofertilizer. The method applied was Biochemical Potential of Methane, through benchtop bioreactors with a volume of 250 mL, and in mesophilic conditions. Waste was used as inoculum from the manure tank of the quail egg production farm. Thesubstrates used to compose the treatments were liquid quail farming waste from the water depth treatment system, with 15, 30 and 45 days of deposition. In the results obtained, it was verified that the best mono digestion used was inoculum+substrate of 30 days of deposition, with water retention time of 45 days, showing a higher production accumulated in biogas (0.00078476 Nm3) and CH4 (0.000575 Nm3) as well as the highest biogas potential of 0.0043 Nm3 (kg substrate)-1. When converted into electrical energy,  by means of a motor generator, using as fuel the biogas produced by the liquid quail farming waste, the value of 104.64 kwh (45 days)-1 was obtained.A coturnicultura, enquanto atividade que demanda baixo investimento e tem rápido retorno, mostra-se como possibilidade para o produtor rural familiar. Concomitantemente a isso, destaca-se o fato de que a utilização dos dejetos líquidos da coturnicultura, visando à geração de energia por digestão anaeróbia, pode significar uma tecnologia viável e promissora para a obtenção de biogás originado dos sistemas de produção de animais confinados. A crescente demanda por energia estabelece que novas fontes energéticas sejam mais aproveitadas, e uma grande oportunidade para o crescimento destas pode ser o uso da biomassa em sistemas de digestão anaeróbia, em que o substrato orgânico é degradado e transformado em energia e biofertilizante. O método aplicado foi o Potencial Bioquímico de Metano, por meio de biorreatores em bancada com volume de 250 mL,e em condições mesófilas se utilizou como inóculo o dejeto do tanque da esterqueira da granja de produção de ovos de codornas, os substratos utilizados para compor os tratamentos foram dejetos líquidos da coturnicultura do sistema de tratamento por lâmina d’água, com 15, 30 e 45 dias de deposição. Nos resultados obtidos, constatou-se que a melhor monodigestão utilizada foi inóculo+substrato de 30 dias de deposição, com tempo de retenção hídrica de 45 dias, apresentando maior produção acumulada de biogás (0,00078476 Nm3) e CH4 (0,000575 Nm3), bem como o maior potencial de biogás 0,0043 Nm3 (kg substrato)-1; e, quando convertido em energia elétrica, por meio de motogerador, utilizando como combustível o biogás produzido pelo dejetos líquidos da coturnicultura, obteve-se o valor 104,64 kwh (45 dias)-1

Study of the occurrence of agrochemicals in the waters of the Marombas River- Santa Catarina - Brazil

The region of the sub-basin of the Marombas River is based on economic agroforestry activities. These activities use agrochemicals that can contaminate surface and groundwater and cause diseases in humans and animals. The present study aimed to detect the presence of agrochemicals in the waters of the Marombas River at 4 different points. The water samples were collected every two months over a 1-year period. The concentrations of carbamates, herbicides, fungicides and organophosphates were analyzed. The chromatographic analysis revealed the presence of the fungicide Azoxystrobin (0.65 μg L-1) in the water sample collected at a point in the river with a strong influence of agricultural activities. The concentration of other agrochemicals was below the detection limit of the assay methods used at all points, and in all types of sampling. The Marombas River is subject to contamination by agrochemicals, which represents a potential risk to the human population and to the aquatic biota

Biogas production and electricity generation from a quail manure wastewater treatment system per water depth

A coturnicultura, enquanto atividade que demanda baixo investimento e tem rápido retorno, mostra-se como possibilidade para o produtor rural familiar. Concomitantemente a isso, destaca-se o fato de que a utilização dos dejetos líquidos da coturnicultura, visando à geração de energia por digestão anaeróbia, pode significar uma tecnologia viável e promissora para a obtenção de biogás originado dos sistemas de produção de animais confinados. A crescente demanda por energia estabelece que novas fontes energéticas sejam mais aproveitadas, e uma grande oportunidade para o crescimento destas pode ser o uso da biomassa em sistemas de digestão anaeróbia, em que o substrato orgânico é degradado e transformado em energia e biofertilizante. O método aplicado foi o Potencial Bioquímico de Metano, por meio de biorreatores em bancada com volume de 250 mL,e em condições mesófilas se utilizou como inóculo o dejeto do tanque da esterqueira da granja de produção de ovos de codornas, os substratos utilizados para compor os tratamentos foram dejetos líquidos da coturnicultura do sistema de tratamento por lâmina d’água, com 15, 30 e 45 dias de deposição. Nos resultados obtidos, constatou-se que a melhor monodigestão utilizada foi inóculo+substrato de 30 dias de deposição, com tempo de retenção hídrica de 45 dias, apresentando maior produção acumulada de biogás (0,00078476 Nm3) e CH4 (0,000575 Nm3), bem como o maior potencial de biogás 0,0043 Nm3 (kg substrato)-1; e, quando convertido em energia elétrica, por meio de motogerador, utilizando como combustível o biogás produzido pelo dejetos líquidos da coturnicultura, obteve-se o valor 104,64 kwh (45 dias)-1

Chemical Composition And Cytotoxic Activity Of The Essential Oils Of Cantinoa Stricta (benth.) Harley & Jfb Pastore (lamiaceae)

The essential oils from the leaves and flowers of Cantinoa stricta (Benth.) Harley & J.F.B. Pastore (Lamiaceae) were obtained by hydrodistillation and analyzed by GC/FID and GC/MS. The major components of both oils were caryophyllene oxide (leaf - 31.6%; flower - 21.7%) and cis-pinane (leaf -15.4%; flower - 9.7%). The flower oil also contained significant amounts of alpha-pinene (9.4%) and beta-pinene (9.1%). The oils were tested in vitro against U251 (glioma), UACC-62 (melanoma), MCF-7 (breast), NCI-H460 (lung), PC-3 (prostate), K-562 (leukemia) human cancer cell lines and against HaCat (no cancer cell), using the sulforhodamine B method. Both oils showed antiproliferative activity against all tested cells lines (TGI < 50 mu g/mL), with exception of K562 cells. The highest activity was observed against MCF-7 cell lines (TGI = 4.54-10.36 mu g/mL).10225726

Chemical composition and cytotoxic activity of the essential oils of Cantinoa stricta (Benth.) Harley & J.F.B. Pastore (Lamiaceae)

The essential oils from the leaves and flowers of Cantinoa stricta (Benth.) Harley & J.F.B. Pastore (Lamiaceae) were obtained by hydrodistillation and analyzed by GC/FID and GC/ MS. The major components of both oils were caryophyllene oxide (leaf – 31.6%; flower – 21.7%) and cis-pinane (leaf – 15.4%; flower – 9.7%). The flower oil also contained significant amounts of a -pinene (9.4%) and b -pinene (9.1%). The oils were tested in vitro against U251 (glioma), UACC-62 (melanoma), MCF-7 (breast), NCI-H460 (lung), PC-3 (prostate), K-562 (leukemia) human cancer cell lines and against HaCat (no cancer cell), using the sulforhodamine B method. Both oils showed antiproliferative activity against all tested cells lines (TGI < 50 m g/mL), with exception of K562 cells. The highest activity was observed against MCF-7 cell lines (TGI = 4.54-10.36 m g/mL).10225726

VOLATILE COMPONENTS FROM GALLS INDUCED BY Baccharopelma dracunculifoliae

The volatile components of the galls induced by the insect Baccharopelma dracunculifoliae (Hemiptera: Psyllidae) on leaves of Baccharis dracunculifolia (Asteraceae) were analyzed by gas chromatography-mass spectrometry (GC-MS) and gas chromatographyflame- ionisation detection (GC-FID), and then comparison with volatile oil samples from healthy leaves collected in the vicinity. The galls produced around 3.5% of the total organic volatiles whereas healthy leaves rendered an average yield of 0.6%. The observed higher proportions of germacrene D, bicyclogermacrene, limonene, and β-pinene in the galls suggest that all these compounds are important targets in the search for natural enemies of this Psyllid. Moreover, higher relative percentages of (E)-nerolidol and spathulenol were found in healthy leaves

Chemical composition, antioxidant and anti-inflammatory activities of the essential oils from male and female specimens of Baccharis punctulata (Asteraceae)

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Chemical composition of volatiles from male and female specimens of Baccharis trimera collected in two distant regions of southern brazil: a comparative study using chemometrics

GC/MS/FID analyses of volatile compounds from cladodes and inflorescences from male and female specimens of Baccharis trimera (Less.) DC. collected in the states of Paraná and Santa Catarina, Brazil, showed that carquejyl acetate was the primary volatile component (38% to 73%), while carquejol and ledol were identified in lower concentrations. Data were subjected to hierarchical cluster analysis and principal component analysis, which confirmed that the chemical compositions of all samples were similar. The results presented here highlight the occurrence of the same chemotype of B. trimera in three southern states of Brazil