A new HPLC-DAD-MS/MS method for the simultaneous determination of major compounds in the crude extract of Lychnophora salicifolia Mart. (Brazilian arnicao) leaves: Application to chemical variability evaluation

Lychnophora salicifolia Mart., which occurs in the Brazilian Cerrado in the states of Bahia and Minas Gerais as well as in the southeast of the state of Goias, is the most widely distributed and also the most polymorphic species of the genus. This plant is popularly known to have anti-inflammatory and analgesic activities. In this work, we have studied the variation in terms of polar metabolites of ninety-three Lychnophora salicifolia Mart, specimens collected from different regions of the Brazilian Cerrado. Identification of the constituents of this mixture was carried out by analysis of the UV spectra and MS data after chromatographic separation. Twenty substances were identified, including chlorogenic acid derivatives, a flavonoid C-glucoside, and other sesquiterpenes. The analytical method was validated, and the reliability and credibility of the results was ensured for the purposes of this study. The concentration range required for analysis of content variability within the analyzed group of specimens was covered with appropriate values of limits of detection and quantitation, as well as satisfactory precision and recovery. A quantitative variability was observed among specimens collected from the same location, but on average they were similar from a chemical viewpoint. In relation to the study involving specimens from different locations, there were both qualitative and quantitative differences among plants collected from different regions of Brazil. Statistical analysis revealed that there is a correlation between geographical localization and polar metabolites profile for specimens collected from different locations. This is evidence that the pattern of metabolites concentration depends on the geographical distribution of the specimens. (C) 2012 Elsevier B.V. All rights reserved.FAPESP [2008/02323-3]FAPESPCNPqCNPqCAPESCAPE

Metabolic response of Adenocalymma peregrinum during regeneration of the aerial parts

Adenocalymma peregrinum, popularly known as ciganinha, is an aggressive pasture invader. This species has a vegetative propagation mechanism that includes aerial latent buds and subterranean stalks activated by cuts and other lesions. In the present study, we evaluated the levels of cytokinins and secondary metabolites in A. peregrinum during regeneration of the aerial part. Plants used in this experiment were established in the field and were cut at intervals of five days until the 25th day. The regeneration of the cut plants started on the 10th day after the first cut. Among observed compounds, only kinetin, allantoin and the iridoide 6-β-hydroxyipolamiide were detected at adequate levels for comparative evaluation. The maximum levels of these compounds coincided with the beginning of regeneration in these plants, indicating the involvement of these compounds in the regeneration process

Seasonal variation of the major secondary metabolites present in the extract of Eremanthus mattogrossensis Less (Asteraceae: Vernonieae) leaves

The species Eremanthus mattogrossensis, known as "veludo do cerrado" (cerrado velvet), is native to the Brazilian Cerrado. Because the amount of metabolites present in plants may be influenced by biological and environmental factors, here we conducted an HPLC-DAD-MS/MS investigation of the metabolite concentrations found in the MeOH/H2O extract of the leaves of this species. The main compounds were identified and quantified, and the metabolites were grouped by chemical class (caffeoylquinic acids, flavonoids, and sesquiterpene lactone). Statistical analysis indicated a straight correlation between the quantity of metabolites and seasonality, suggesting that environmental properties elicit important metabolic responses

Projeto ExPOSE: criação de protocolos para avaliação da exposição ocupacional ao microbiota em ambientes clínicos

FCT - 02/SAICT/2016 – Project nº 23222Nas últimas décadas, tem sido enfatizada a importância dos bioaerossóis em ambientes interiores, pelos efeitos adversos que causam na saúde humana e consequente impacto na saúde pública. Os hospitais e restantes unidades de saúde, pelas caraterísticas inerentes aos mesmos podem potenciar a contaminação microbiana, pelo que requerem uma atenção e intervenção especial para proteger, não só os pacientes mas também os profissionais de saúde, das infeções associadas aos cuidados de saúde e das doenças profissionais. O projeto ExPOSE tem como objetivo avaliar a exposição dos trabalhadores ao microbiota em unidades de cuidados de saúde e consequente elaboração de orientações para a redução dessa exposição.info:eu-repo/semantics/publishedVersio

Sensitive method for determination of piplartine, an alkaloid amide from Piper species, in rat plasma samples by liquid chromatography-tandem mass spectrometry

Piplartine (PPTN) is an alkaloid amide found in Piper species that presents different activities. PPTN determination in rat plasma is necessary to better understand its biological effects. The aim of this study was to develop a sensitive LC-MS/MS method for the determination of PPTN in rat plasma. The performance criteria for linearity, sensitivity, precision, accuracy, recovery, and stability have been assessed and were within the recommended guidelines. The validated method proved to be suitable in a pilot study of PPTN kinetic disposition in rat plasma after a single intraperitoneal dose, and represents an appropriate tool to further pharmacokinetic studies

In vitro and in vivo metabolism of the major secondary metabolites present in the extract of Lychnophora salicifolia Mart. (Asteraceae: Vernonieae)

A espécie Lychnophora salicifolia Mart. é conhecida popularmente como arnicão e é utilizada como anti-inflamatório e analgésico de uso tópico, além de edulcorante em cachaça. A vicenina-2 é um flavonoide C-glicosilado presente em grande quantidade no extrato hidroalcoólico das folhas e apresenta ação anti-inflamatória, antioxidante, anticâncer entre outras. Outra substância majoritária nesse extrato, o ácido lychnofólico, é um dos responsáveis pela ação antibacteriana e tripanocida do extrato. Devido à necessidade de estudos mais completos envolvendo produtos naturais, neste trabalho estudamos o metabolismo dessas duas moléculas sendo que não foi possível observar a formação de metabólitos in vitro a partir da vicenina-2 e no caso do ácido lychnofólico houve a formação de dois metabólitos pela adição de um átomo de oxigênio na molécula utilizando microssomas hepáticos de ratos. As mesmas substâncias foram formadas utilizando FeTFPP como catalisador e m-CPBA como agente oxidante. Nas reações com microssomas hepáticos humanos foi observada a conjugação do ácido lychnofólico com o ácido glucurônico sendo possível calcular: Vmáx = 272,5 ± 12,75 ?mol/g proteína/hora; Km = 12,38 ±1,218 ?M e o R2 = 0,9980. Os estudos com células Caco-2 indicam que o ácido lychnofólico é rapidamente absorvido por difusão passiva. Já a vicenina-2 em nenhuma condição avaliada atravessou a monocamada de células. Quanto ao perfil farmacocinético do ácido lychnofólico verificou-se um baixíssimo t1/2, alto Cl e alto Vd. Já a vicenina-2 apresentou valores um pouco maiores, mas ainda sim, baixos de t1/2 e Cl e Vd, indicando que ambas as substâncias são rapidamente eliminadas do organismo.The species Lychnophora salicifolia Mart. popularly known as \"arnicão\" is used as an antiinflammatory and topical analgesic agent; it is also employed as flavoring in the Brazilian cachaça. Vicenin-2 is a C-glycoside flavonoid present in large amounts in the hydroalcoholic extract of the leaves of L. salicifolia and has anti-inflammatory, antioxidant, and anticancer activities. Another major substance in this extract is lychnopholic acid, which accounts for the antibacterial and trypanocidal actions of the extract. More comprehensive studies involving natural products are necessary; therefore, we studied the metabolism of the two aforementioned molecules. We did not observe the in vitro formation of metabolites from vicenin-2. In the case of lychnopholic acid, we detected the formation of two metabolites using rat liver microsomes, which possibly originated from the addition of an oxygen atom to the acid\'s molecule. The same substances may also have been produced when we employed FeTFPP as catalyst and m-CPBA as oxidizing agent. In the reactions involving human liver microsomes, we verified that lychnopholic acid conjugation with glucuronic acid and calculated the following parameters: Vmax = 272.5 ± 12.75 ?mol g-1 protein h-1; Km = 12.38 ± 1.218 ?M, and R2 = 0.9980. Studies using Caco-2 cells indicated that lychnopholic acid was quickly absorbed by passive diffusion. On the other hand, vicenin-2 was not transported, suggesting no absorption or efflux of this compound in Caco-2 cells. As for the pharmacokinetic profile, lychnopholic acid presented very low t1/2, high Cl, and very high Vd. Concerning vicenin-2, the t1/2 value was slightly larger, but still rather low, whereas Cl and Vd values were slightly lower, indicating that both substances are quickly eliminated from the body

Interpopulational variation of secondary metabolites of arnicão (Lychnophora salicifolia Mart., Vernonieae, Asteraceae).

Cerca de 10% da flora mundial é representada pela família Asteraceae, uma das maiores famílias de Angiospermae, caracterizada não só pela diversidade morfológica e taxonômica, como também pela riqueza de metabólitos secundários apresentada pelas espécies estudadas dessa família. Lychnophora salicifolia Mart. é a espécie de maior distribuição geográfica do gênero (ocorrendo na Serra do Espinhaço na Bahia, em Minas Gerais e no sudeste de Goiás) e a mais polimórfica também. Entre seus usos etnobotânicos, destaca-se a ação anti-inflamatória e analgésica. Neste trabalho, realizamos o estudo da variação dos metabólitos polares dessa espécie, sendo que para isso foi utilizada a desreplicação do extrato bruto (metanol/água obtido a partir da folhas) através da análise por CLAE-DAD-EM, a qual permitiu a caracterização dos constituintes dessa mistura através da análise dos espectros de UV e dados de EM. Assim foi possível identificar 20 substâncias: ácido protecatecuico; ácido 3-O-E-cafeoilquínico; ácido 3-O-p-cumaroilquínico; ácido 4-O-E-cafeoilquínico; ácido 5-O-E-cafeoilquínico; ácido 4-O-p-cumaroilquínico; ácido 5-O-p-cumaroilquinico; ácido 5-O-feruloilquínico; vicenina-2 (6,8-di-C--glcosilapigenina); ácido 3,4-di-O-E-cafeoilquínico; ácido 3,5-di-O-E-cafeoilquínico; ácido 3-O-cumaroil 5-O-p-cafeoilquínico; ácido 3-O-cafeoil, 5-O-p-cumaroilquínico; ácido 4,5-di-O-E-cafeoilquínico; ácido 4-feruloil-5-O-E-cafeoilquínico; ácido 3,4 di-O-p-cumaroilquínico; ácido 3,5 di-O-p-cumaroilquínico; ácido 4,5 di-O-p-cumaroilquínico; ácido lychnofólico e o ácido 2-O-acetil lychnofólico. Para a validação da metodologia analítica foi utilizada uma substância pertencente a cada classe de compostos: ácido 5-O-E-cafeoilquínico, vicenina-2 e o ácido 2-O-acetil lychnofólico. Quanto à validação da metodologia analítica, esta garantiu a confiabilidade dos resultados e a credibilidade do método para os propósitos deste estudo, abrangendo a faixa de concentração necessária para as análises de variação populacional, além apresentar valores de limites de detecção e limites de quantificação, precisão e recuperação adequados. Na análise intrapopulacional, verificou-se uma variabilidade quantitativa entre indivíduos da mesma população, porém, em média os indivíduos pertencentes à mesma população são parecidos quimicamente. Já em relação ao estudo da variação populacional, pudemos verificar diferenças tanto quanti como qualitativas entre populações coletadas em regiões distintas do Brasil, sendo que a partir da análise estatística, verificamos que há uma correlação entre isolamento geográfico e diminuição da semelhança do perfil de metabólitos polares entre as plantas de populações diferentes indicando que o padrão encontrado para concentração dos metabólitos é influenciado pela distribuição geográfica das amostras. Todavia, não é possível determinar a localização das amostras unicamente pelas variáveis metabólicas estudadas. Assim, a concentração e presença dos metabólitos são influenciadas pela distribuição geográfica das amostras.About 10% of the world flora is represented by the Asteraceae family, which has been extensively studied since it is one of the largest families of Angiospermae, characterized not only by morphological and taxonomic diversity but also the wealth of secondary metabolites produced by the species studied of this family. Lychnophora salicifolia Mart. is the most widely distributed species of the genus (occurring in the Serra do Espinhaço in Bahia, Minas Gerais and in the southeast of Goiás) and also is the most polymorphic. Its popular use is due to the anti-inflammatory and analgesic activity. In this work, we studied the variation of polar metabolites of this species using the dereplication of the crude extract (methanol/water obtained from the leaves) by the HPLC-DAD-MS analysis, which led us to identify the constituents of this mixture by analyzing the UV spectra and MS data. Thus it was possible to identify 20 substances: protocatechuic acid; 3-O-E-caffeoylquinic acid; 3-O-p-coumaroylquinic acid; 4-O-E-caffeoylquinic acid; 5-O-E-caffeoylquinic acid; 4-O-p-coumaroylquinic acid; 5-O-p-coumaroylquinic acid; 5-O-feruloylquinic acid; vicenin-2 (6,8-di-C--glucosylapigenin); 3,4-di-O-E-caffeoylquinic acid, 3,5-di-O-E-caffeoylquinic acid; 3-O-coumaroyl-5-O-p-caffeoylquinic acid; 3-O-caffeoyl acid, 5-O-p-coumaroylquinic acid; 4,5-di-O-E-caffeoylquinic acid, 4-feruloyl-5-O-E-caffeoylquinic acid; 3,4-di-O-p-coumaroylquinic acid; 3,5-di-O-p-coumaroylquinic acid; 4,5-di-O-p-coumaroylquinic acid; lychnofolic acid and 2-O-acetyl lychnofolic acid. To the validation of analytical methodology were used one substance belonging to each class of compounds: 5-O-E-caffeoylquinic acid, vicenin-2 and 2-O-acetyl lychnofolic acid. So, the validation of analytical method ensured the reliability and credibility of the results of the method for purposes of this study, covering the concentration range required for analysis of population variation and presents appropriates values of detection and quantification limits, precision and recovery. For intrapopulation analysis, there was a quantitative variability between individuals of the same population, but on average the individuals of the same population are similar chemically. In relation to the study of interpopulation variation, we could see quantitative and qualitative differences between populations collected from different regions of Brazil, and from the statistical analysis, we found that there is a correlation between geographic isolation and decrease of the similarity of the polar metabolites profile between plants of different populations. This indicated that the pattern found for the concentration of metabolites is influenced by the geographical distribution of samples. However, it is not possible to determine the location of the samples only by the metabolic variables studied. Thus, the presence and concentration of metabolites are influenced by the geographical distribution of samples

The Protective Role of Lychnophora ericoides Mart. (Brazilian Arnica) in 1,2-Dimethylhydrazine-Induced Experimental Colon Carcinogenesis

Aberrant crypt foci (ACF) and colon rectal mucosal epithelial cell proliferation have been shown to be increased in patients with colon cancer and have been largely used for early detection of factors that influence colorectal carcinogenesis in rats. Fifty male Wistar rats were randomly divided into 5 groups. The groups G1 to G4 were given 4 injections of the carcinogen 1,2-dimethylhydrazine (DMH). The G2 group received Lychnophora ericoides (LE) extracts for 6 wk. The groups G3 and G4 received LE for 4 wk and 2 wk, respectively, at the postinitiation and initiation phases of colonic carcinogenesis. The group G5 was the control. Forty-two days after the first injections of DMH for the neoplasic induction, we observed a statistically significant decrease in the number of aberrant crypt foci (ACF) and an attenuation of the increase in cell proliferation induced by DMH in all the LE-treated groups. Thus, we concluded that Lychnophora ericoides extracts were effective against the development of cancer. These data suggest that LE has a protective influence on the process of colon carcinogenesis, suppressing both the initiation and the promotion of colonic carcinogenesis.CAPESCNPq[Proc.475844/2004-1]FAPESP[2001/14227-0