Characterization and Quantification of Compounds in the Hydroalcoholic Extract of the Leaves from Terminalia catappa

Terminalia is a genus of Combretaceous plants widely distributed in tropical and subtropical regions. Thus, the aim of this study was to quantify the majority compounds of the hydroalcoholic extract (7 : 3, v/v) of the leaves from T. catappa by HPLC-PDA, chemically characterize by hyphenated techniques (HPLC-ESI-IT-MSn) and NMR, and evaluate its mutagenic activity by the Salmonella/microsome assay on S. typhimurium strains TA98, TA97a, TA100, and TA102. The quantification of analytes was performed using an external calibration standard. Punicalagin is the most abundant polyphenol found in the leaves. The presence of this compound as a mixture of anomers was confirmed using HPLC-PDA and 1H and 13C NMR. Mutagenic activity was observed in strains TA100 and TA97a. As the extract is a complex mixture of punicalagin, its derivatives, and several other compounds, the observed mutagenicity may be explained in part by possible synergistic interaction between the compounds present in the extract. These studies show that mutagenic activity of T. catappa in the Ames test can only be observed when measured at high concentrations. However, considering the mutagenic effects observed for T. catappa, this plant should be used cautiously for medicinal purposes

Avaliação da mutagenicidade, antimutagenicidade e estrogenicidade de Byrsonima spp

O cerrado brasileiro contém inúmeras plantas utilizadas no tratamento popular de várias enfermidades onde destacamos as espécies do gênero Byrsonima. Os relatos de seu uso são diversos: tratamento de infecções fúngicas e bacterianas, feridas crônicas, doença de Chagas, contra tuberculose, como diurética, antiemética e etc. Na literatura, há comprovação para atividades: antioxidante, antiprotozoário, depressor no Sistema Nervoso Central (SNC), antihiperglicêmicas e antihiperlipêmica. O perfil fitoquímico demonstra taninos, flavonóides, triterpenos, ésteres aromáticos, entre outros, o que indica o potencial farmacológico deste gênero. Sabe-se que o uso não padronizado de plantas pode ser danoso aos usuários, devido a propriedades desconhecidas ou por conterem contaminantes ambientais. Embora haja diversos estudos abordando o potencial biológico das plantas deste gênero, amplamente utilizadas na medicina popular, ainda há poucos estudos sobre sua segurança Por esta razão, o objetivo deste trabalho foi avaliar as atividades mutagênica, antimutagência e estrogênica de extratos etanólicos padronizados (70%) de folhas de quatro espécies: Byrsonima verbascifolia, Byrsonima coccolobifolia, Byrsonima correifolia e Byrsonima ligustrifolia. A atividade mutagênica foi realizada pelo Teste de Ames, utilizando o ensaio de pré-incubação, na ausência e presença de ativação metabólica (S9). As linhagens utilizadas foram TA98, TA97a, TA100 e TA102 e foram testadas cinco concentrações de cada amostra. A atividade antimutagênica foi testada apenas para as amostras negativas para o teste de Ames (B. verbascifolia e B. correifolia), com a diferença que sempre foi associada a um agente mutagênico conhecido. A atividade estrogênica foi avaliada por dois testes: -Recombinant Yeast Assay (RYA) que utiliza a levedura Saccharomyces cerevisiae (linhagem BY4741) modificada geneticamente, contendo um gene para receptor e...The Brazilian cerrado contain several native plants used in folk medicine for treating many diseases. Plants of the genus Byrsonima are one of them. There are several reports of its use: the treatment of fungal and bacterial infections, chronic wounds, Chagas disease, tuberculosis, as a diuretic and antiemetic. In literature there is evidence for activities: antioxidant, antiprotozoal, depressing CNS, antihiperglicemic and antihiperlipemic. The phytochemical profile shows tannins, flavonoids, triterpenes, aromatic esters and others, which indicates the pharmacological potential of this genus. It is known that the use of non-standard plant can be harmful to users due to unknown properties or for containing environmental contaminants. Although there are several studies about the biological potential of plants of this genus, widely used in folk medicine, there are few studies about their safety. For this reason, the aim of this study was to investigate the mutagenic, antimutagenic and estrogenic activities of standardized ethanolic extracts (70%) of leaves of four species: Byrsonima verbascifolia, Byrsonima correifolia, Byrsonima coccolobifolia and Byrsonima ligustrifolia. Mutagenic acivity was evaluated by Ames Test, performed according to preincubation assay, in absence and presence of metabolic activation system (S9). TA98, TA97a, TA100 and TA102 strains were used and five concentrations of each sample, in triplicates, were tested. For antimutagenicity assay were used only the samples negative in Ames Test (B. verbascifolia and B. correifolia), with the difference that samples were associated a known mutagenic agent The estrogenic activity was evaluated by two assays: -The Recombinant Yeast Assay (RYA) uses the yeast Saccharomyces cerevisiae (BY4741 strain)genetically modified, containing a gene for human estrogen receptor and a plasmid which expresses the β-galactosidase enzyme of Escherichia coli and - Escreen, a test that evaluates ...Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Estrogenic and mutagenic activities of crotalaria pallida measured by recombinant yeast assay and ames test

Background: Crotalaria pallida Ailton is a plant belonging to the Fabaceae family, popularly known as rattle or rattlesnake and used in traditional medicine to treat swelling of the joints and as a vermifuge. Previous pharmacological studies have also reported anti-inflammatory, antimicrobial and antifungal activities. Nevertheless, scientific information regarding this species is scarce, and there are no reports related to its possible estrogenic and mutagenic effects. Thus, the purpose of the present study was to investigate the estrogenic potential of C. pallida leaves by means of the Recombinant Yeast Assay (RYA), seeking an alternative for estrogen replacement therapy during menopause; and to reflect on the safe use of natural products to assess the mutagenic activity of the crude extract from C. pallida leaves, the dichloromethane fraction and stigmasterol by means of the Ames test.Methods: The recombinant yeast assay with the strain BY4741 of Saccharomyces cerevisiae, was performed with the ethanolic extract, dichloromethane fraction and stigmasterol isolated from the leaves of C. pallida. Mutagenic activity was evaluated by the Salmonella/microsome assay (Ames test), using the Salmonella typhimurium tester strains TA100, TA98, TA97 and TA102, with (+S9) and without (-S9) metabolization, by the preincubation method.Results: All samples showed estrogenic activity, mainly stigmasterol. The ethanolic extract from C. pallida leaves showed mutagenic activity in the TA98 strain (-S9), whereas dichloromethane fraction and stigmasterol were found devoid of activity.Conclusion: Considering the excellent estrogenic activity performed by stigmasterol in the RYA associated with the absence of mutagenic activity when evaluated by the Ames test, stigmasterol becomes a strong candidate to be used in hormone replacement therapy during menopause. © 2013 Boldrin et al.; licensee BioMed Central Ltd