The Cannabis sativa L. plant has a complex chemical composition, containing various chemical compounds such as terpenes, sugars, hydrocarbons, steroids, flavonoids, and amino acids. Few works have attempted to identify the constitutional isomers of cannabinoids that are found in the plant. The present study reported the analysis of seven cannabinoid standards: five neutral and two acidic, as well as Cannabis products (hashish and marijuana) and parts of the Cannabis plant (flower and leaf) using mono-dimensional gas chromatography coupled with mass spectrometry (GC-MS) and two-dimensional gas chromatography coupled with quadrupole MS (GC × GC-QMS), ultra performance liquid chromatography coupled with electrospray ionization-quadrupole-time of flight (UPLC-ESI-QTOF)-MS and UPLC-ESI-travelling wave ion mobility (TWIM)-MS. The results of GC-MS demonstrated close retention times (∆tR = 1.303 min) in separation of the five cannabinoid standards, whereas GC × GC-QMS provided a substantially better identification and distinction of constitutional isomers of cannabinoids, where a total of 11 cannabinoids were identified in the hashish sample. UPLC-QTOF-MS and UPLC-TWIM-MS data obtained complete chemical information, in which ESI(+) revealed the presence of seven constitutional isomers of ∆9-tetrahydrocannabinol (∆9-THC), whereas ESI(-) proved the presence of four isomers of ∆9-tetrahydrocannabinolic acid A (∆9-THCA A).

Abstract In hybrid cultivated form, Dendranthema grandiflorum (Ramat.) Kitam., Asteraceae, flowers (Chrysanthemum morifolium Ramat.) were utilized in the production of extracts, which were analyzed for larvicidal activity against Aedes aegypti third instar larvae. Methanol and dichloromethane extracts showed LC50 values of 5.02 and 5.93 ppm, respectively. Using GC–MS, phytochemical analyses of the dichloromethane extract showed the presence of triterpenoids and fatty acids, while flavonoids and caffeoylquinic acids were shown to occur in the methanol extract by ESI Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (ESI-FT-ICR-MS). Triterpenoids and fatty acids are well known insecticidal compounds. From this study, it can be concluded that D. grandiflorum grown for floriculture, as an agribusiness, can have additional applications as raw material for the production of insecticidal products.

<p>This paper describes the correlation between the phenolic composition and the molecular phylogenetic reconstruction of five <italic>Spondias</italic> species (Anacardiaceae). Two of these species (<italic>S. venulosa</italic> and <italic>Spondias</italic> sp.) occur in rainforest areas and the other three are widely distributed in Brazil (<italic>S. dulcis</italic>, <italic>S.mombin</italic>, and <italic>S. purpurea</italic>). The flavonoid enriched fraction of the <italic>S. venulosa</italic> leaf extract also underwent a chemical study. The results indicate that the presence of flavonol 3-<italic>O</italic>-glycosides are a synapomorphic character of the studied American <italic>Spondias</italic> and the production of rhamnetin 3-<italic>O</italic>-rutinoside is a synapomorphy of the Atlantic forest species. This is the first report of flavonoids in <italic>S. venulosa</italic>, an endemic species from the Brazilian Atlantic rainforest.</p>

Extratos de Echinodorus grandiflorus, obtidos a partir de folhas secas por três técnicas diferentes, foram avaliados pelo ensaio de indução lisogênica bacteriana (Induteste), em relação às propriedades genotóxicas. As amostras foram previamente esterilizadas por vapor de água (autoclavação) ou luz ultravioleta. Somente os extratos preparados por infusão e esterilizados a vapor mostraram atividade genotóxica. A análise fitoquímica revelou a presença dos flavonóides isovitexina, isoorientina, swertisina e swertiajaponina. Isovitexina e isoorientina, isoladas de uma fração genotóxica, foram testadas separadamente e não apresentaram genotoxidade. O desenvolvimento de odor adocicado e escurecimento dos extratos submetidos ao calor motivaram a investigação quanto às substâncias precursoras da reação de Maillard no extrato. Ácidos aminados e açúcares redutores foram identificados e a presença de precursores da reação de Maillard no extrato foi confirmada por espectrofotometria no visível com a adição do inibidor bissulfito de sódio antes do aquecimento. Este é o primeiro relato de aparecimento de melanoidinas em uma técnica que pode ser usada para a preparação de fitoterápico. A atividade genotóxica de tais infusões preparadas ao calor implica em algum risco de desenvolvimento de doenças degenerativas com seu uso descontrolado e por longo prazo.

Extracts of Echinodorus grandiflorus obtained from dried leaves by three different techniques were evaluated by bacterial lysogenic induction assay (Inductest) in relation to their genotoxic properties. Before being added to test cultures, extracts were sterilized either by steam sterilization or ultraviolet light. Only the extracts prepared by infusion and steam sterilized have shown genotoxic activity. The phytochemical analysis revealed the presence of the flavonoids isovitexin, isoorientin, swertisin and swertiajaponin, isolated from a genotoxic fraction. They were assayed separately and tested negative in the Inductest protocol. The development of browning color and sweet smell in extracts submitted to heat, prompted further chemical analysis in search for Maillard's reaction precursors. Several aminoacids and reducing sugars were cast in the extract. The presence of characteristic Maillard's melanoidins products was determined by spectrophotometry in the visible region and the inhibition of this reaction was observed when its characteristic inhibitor, sodium bisulfite, was added prior to heating. Remarkably, this is the first paper reporting on the appearance of such compounds in a phytomedicine preparation under a current phytopharmaceutical procedure. The genotoxic activity of such heat-prepared infusions imply in some risk of developing degenerative diseases for patients in long-term, uncontrolled use of such phytomedicines.

The ethanol extract of the vegetal species Pentaclethra macroloba (Willd.) Kuntze, Fabaceae, was fractioned and the antibacterial activity was determined. The active ethyl acetate (ea) fraction showed activity against Gram-positive (Staphylococcus spp. and Enterococcus spp.) and Gram-negative (Pseudomonas aeruginosa, Acinetobacter spp. and Klebsiella pneumoniae) multiresistant bacteria. Gallic acid derivatives were identified as the main compounds in inactive subfractions from the ea fraction, while the active one afforded ellagic acid as the major constituent when submitted to acid hydrolysis reaction, which suggests the presence of hydrolysable tannins. The minimum bactericidal concentration analysis showed a bactericide mechanism of action for the tannin subfraction found. The antibacterial mechanism of action of the active tannin subfraction against S. aureus reference strains (ATCC 29213 e 33591) was proposed adopting an in vitro assay of protein synthesis inhibition. For this, bacterial cells were labeled with [35S] methionine in the presence of the subfraction. The protein synthesis inhibition was observed at 256 µg/mL of this subfraction. At this concentration it did not present cytotoxicity in eukaryotic cells by the neutral red technique, suggesting selective toxicity. The present study is the first in vitro investigation of the antibacterial properties of tannin fractions obtained from a polar extract of P. macroloba.

In order to validate the Bumelia sartorum Mart., Sapotaceae, traditional use for infection diseases, this study evaluates the antibacterial activity of the stem bark fractions against methicillin-sensitive (MSSA) and methicillin-resistant (MRSA) Staphylococcus aureus strains by using the agar dilution method and reported as MIC (minimal inhibitory concentration). In addition, the DPPH scavenging activity of these fractions was measured and the chemical composition and acute toxicity of the active fraction were also determined. The ethyl acetate (EtOAc) extract was chemically analyzed by LC/MS, direct ionization APCI/MS, ¹H NMR and 13C-NMR. All fractions, except butanol extract, presented high antioxidant activity, especially the methanol and the EtOAc extracts, which showed EC50 values (5.67 and 5.30 µg/mL, respectively) considerably lower than the Gingko-standard EGb 761® (38.58 µg/mL). The antibacterial activity against S. aureus strains was observed in EtOAc (MIC 256-512 µg/mL), which showed a very low toxicity. The chemical study of this fraction revealed the abundant presence of polyphenolic compounds. The antibacterial and antioxidant activities reported in this paper for EtOAc extract from B. sartorum and the low toxicity of this fraction opens the possibility that it could be helpful for the developing of new antibacterial agents for treating S. aureus infections.

O presente estudo teve como objetivo verificar a melhor metodologia de extração para rápida e eficiente obtenção de flavonóides a partir de Alpinia zerumbet. Folhas secas foram extraídas com água destilada e etanol 70%, utilizando as metodologias de extração: maceração sob agitação, ultrassom, microondas e agitador. Para verificação dos flavonóides rutina e kaempferol-3-O-glicuronídeo foram utilizadas as técnicas de CCD e CLAE em fase reversa. O solvente etanol 70% foi mais eficiente como extrator. Para as metodologias ultrassom, microondas e agitador, não houve variação significativa para o rendimento utilizando etanol 70% (11 a 14%). A concentração relativa de rutina e kaempferol-3-O-glicuronídeo, respectivamente, foi maior pelos métodos de extração por ultrassom (1,5 e 5,62 mg g-1 folha seca) e microondas (1,0 e 6,64 mg g-1 folha seca), utilizando etanol 70%. Procedimentos rápidos e simplificados de extração otimizam o trabalho fitoquímico e a obtenção de metabólitos secundários.

The current study aims to verify the best method for a rapid and efficient extraction of flavonoids from Alpinia zerumbet. Dried leaves were extracted using distillated water and ethanol 70% by extraction methods of shaking maceration, ultrasonic, microwave and stirring. By the application of TLC and reversed-phase HPLC techniques the rutin and kaempferol-3-O-glucuronide were detected. Ethanol 70% was more efficient for flavonoids extraction than water. No significant yielding variation was verified for ultrasonic, microwave and stirring methods using ethanol 70% (11 to 14%). Relative concentration of rutin and kaempferol-3-O-glucuronide, respectively, was higher by ultrasonic (1.5 and 5.62 mg g-1 dried leaves) and by microwave (1.0 and 6.64 mg g-1 dried leaves) methods using 70% ethanol. Rapid and simplified extraction proceeding optimize phytochemical work and acquisition of secondary metabolites.

Echinodorus macrophyllus, commonly known as chapéu-de-couro, is a medicinal plant used in folk medicine to treat inflammation and rheumatic diseases. In this work, we used short-term bacterial assays based on the induction of SOS functions to examine the genotoxicity and mutagenicity of an aqueous extract of E. macrophyllus leaves. Whole extract and an ethyl acetate fraction showed similar genotoxicity and caused an ~70-fold increase in lysogenic induction. The extract also gave a positive result in the SOS chromotest with an increase of 12-fold in β-Galactosidase enzymatic units. There was a strong trend towards base substitutions and frameshifts at purine sites in the mutations induced by the extract in Escherichia coli (CC103 and CC104 strains) and Salmonella typhimurium test strains (22-fold increase in histidine revertants in TA98 strain). Since reactive oxygen species may be implicated in aging process and in degenerative diseases, we used antioxidant compounds as catalase, thiourea and dipyridyl in the lysogenic induction test. All this compounds were able to reduce the induction factor observed in the treatment with chapéu-de-couro, thus suggesting that the genotoxicity and mutagenicity were attributable to the production of reactive oxygen species that targeted DNA purines.

O presente estudo teve como objetivo verificar a melhor metodologia de extração para rápida e eficiente obtenção de flavonóides a partir de Alpinia zerumbet. Folhas secas foram extraídas com água destilada e etanol 70%, utilizando as metodologias de extração: maceração sob agitação, ultrassom, microondas e agitador. Para verificação dos flavonóides rutina e kaempferol-3-O-glicuronídeo foram utilizadas as técnicas de CCD e CLAE em fase reversa. O solvente etanol 70% foi mais eficiente como extrator. Para as metodologias ultrassom, microondas e agitador, não houve variação significativa para o rendimento utilizando etanol 70% (11 a 14%). A concentração relativa de rutina e kaempferol-3-O-glicuronídeo respectivamente foi maior pelos métodos de extração ultrassom (1,5 e 5,62 mg g-1 folha seca) e microondas (1,0 e 6,64 mg g-1 folha seca) utilizando etanol 70%. Procedimentos rápidos e simplificados de extração otimizam o trabalho fitoquímico e a obtenção de metabólitos secundários.

The current study aims to verify the best method for a rapid and efficient extraction of flavonoids from Alpinia zerumbet. Dried leaves were extracted using distillated water and ethanol 70% by extraction methods of shaking maceration, ultrasonic, microwave and stirring. By the application of TLC and reversed-phase HPLC techniques the rutin and kaempferol-3-O-glucuronide were detected. Ethanol 70% was more efficient for flavonoids extraction than water. No significant yielding variation was verified for ultrasonic, microwave and stirring methods using ethanol 70% (11 to 14%). The relative concentration of rutin and kaempferol-3-O-glucuronide, respectively, was higher by ultrasonic (1.5 and 5.62 mg g-1 dried leaves, respectively) and by microwave (1.0 and 6.64 mg g-1 dried leaves) methods using ethanol. Rapid and simplified extraction proceeding optimize phytochemical work and acquisition of secondary metabolites.

A espécie Calceolaria chelidonioides (Scrophulariaceae), até então inédita nas citações científicas, foi estudada sob o ponto de vista farmacológico buscando-se identificar possíveis atividades antimicrobiana e antioxidante em metodologia in vitro. As partes aéreas dessa espécie demonstraram atividade antioxidante em modelo usando o radical livre DPPH. As flores de C. chelidonioides mostraram grande potencial antibacteriano frente à bactéria Staphylococcus aureus resistente a meticilina MRSA, um dos principais responsáveis em casos de infecção hospitalar.

The species Calceolaria chelidonioides (Scrophulariaceae), not scientific described so for, was studied in pharmacological aspects aiming to identify some anti-microbial and antioxidant activity. The aerial parts showed antioxidant activity using in vitro DPPH model. The flowers from C. chelidonioides showed strong antibacterial potential against meticiline resistant Staphylococcus aureus (MRSA) strains the main responsible for hospital infection complications.

Do extrato butanólico de Croton campestris St. Hill. (Euphorbiaceae) foram isolados quatro flavonóides, todos O-glicosídeos da quercetina. Estas substâncias foram identificadas como 3-O-b-D-apiofuranosil-(1®2)-galactopiranosil quercetina (1), 3-O-b-D-galactopiranosil quercetina (hiperina) (2), 3-O-a-L-arabinopiranosil quercetina (guaijaverina) (3) e 3-O-a-L-ramnopiranosil quercetina (quercitrina) (4).O presente trabalho relata a presença destas substâncias pela primeira vez para esta espécie de Croton, cuja elucidação estrutural deu-se por espectroscopia em UV, EM e RMN, incluindo as técnicas bidimensionais: ¹H-¹H (2D NOESY, 2D COSY), ¹H-13C (2D HETCOR) e 13C (APT), além de comparações com os dados da literatura.

Four flavonoids were isolated from the butanolic extract of the aerial parts of Croton campestris St. Hill. (Euphorbiaceae). These compounds were identified as 3-O-b-D-apiofuranosyl-(1®2)-galactopyranoside quercetin (1), 3-O-b-D-galactopyranoside quercetin (hyperin) (2), 3-O-a-L-arabinopyranoside quercetin (guaijaverin) (3) and 3-O-a-L-ramnopyranoside quercetin (quercitrin) (4). They have been isolated for the first time from Croton campestris. Their structures were elucidated by UV, MS and NMR experiments including ¹H-¹H (2D NOESY, 2D COSY), ¹H-13C (2D HETCOR), 13C (APT) and by comparison of the spectral data with those reported in the literature.

O fracionamento do extrato acetato de etila de frutos de Punica granatum, guiado por ensaios antimicrobianos frente a colônias de Staphylococcus aureus resistentes à meticilina, conduziu ao isolamento e à determinação estrutural do tanino elágico punicalagina. A identificação da substância foi realizada por CLAE/UV e RMN de ¹H. Os ensaios antimicrobianos foram realizados pelo método de difusão em discos de papel. A concentração mínima inibitória das substâncias foi determinada pelo método de diluição em agar padronizada pelo NCCLS (National Committee for Clinical Laboratory Standards).

The ethyl acetate extract of Punica granatum fruits was fractionated by chromatographic techniques to afford the ellagitannin punicalagin. The substance was found to be active against methicillin-resistant Staphylococcus aureus strains and was identified by HPLC/UV and ¹HNMR. The antibacterial assays which guided the isolation of the tannin were conducted using the disc diffusion method. Minimum inhibitory concentration (MIC) was determined by the dilution method according to NCCLS (National Committee for Clinical Laboratory Standards) procedure.