RESUMO
The inflorescences of the Mexican gordolobo are used as a folk medicine to treat various respiratory diseases. Currently, the botanical species that bear the name Mexican gordolobo belong to the genera Gnaphalium and Pseudognaphalium. Despite a long history of traditional use, most Mexican gordolobo species have never been fully chemically characterized, and the range of constituents in the species has not been comprehensively reported. To establish a quality control and chemical characterization method, a total of 49 samples belonging to 18 species of Pseudognaphalium and four species of Gnaphalium were studied. Nine flavones were quantified using a UPLC-PDA method. The method was validated in terms of linearity (R2 > 0.99), precision (intra- and inter-day: 0.1-3.9%), accuracy (96-103%), detection limit (10â¯ng/mL), limit of quantification (25â¯ng/mL) and robustness. 3-Methylquercetin, luteolin, quercetin, 3,5-dihydroxy-6,7,8-trimethoxyflavone, apigenin and gnaphaliin A were present at relatively high levels in most of the samples analyzed. The samples of P. oxyphyllum and P. liebmannii showed the highest content of the 9 compounds analyzed. Whereas the samples of the 5 species of Gnaphalium showed the lowest levels, including non-detectable, of the 9 compounds quantified. This marks an important difference with Pseudognaphalium species. Furthermore, using UHPLC-ESI-QToF data with targeted and non-targeted approaches, 57 compounds, were identified in Mexican gordolobo samples. Flavonoids were the main group of compounds found in Mexican gordolobo.
Assuntos
Flavonas , Gnaphalium , Extratos Vegetais , Cromatografia Líquida de Alta Pressão/métodos , Flavonas/análise , Flavonas/química , Gnaphalium/química , Extratos Vegetais/química , Extratos Vegetais/análise , Limite de Detecção , Reprodutibilidade dos Testes , México , Controle de Qualidade , Medicina Tradicional/métodos , Espectrometria de Massas em Tandem/métodos , Espectrometria de Massas/métodosRESUMO
Aleurites moluccanus (candlenut) and Bertholletia excelsa (Brazil nut) are marketed as dietary supplements for weight loss. These dietary supplements have been found to sometimes be adulterated with toxic nuts/seeds from Cascabela thevetia, commonly known as yellow oleander or lucky nut. This study emphasizes the key identification parameters to differentiate the genuine and adulterated nuts. Samples were obtained from authenticated sources of the nuts and from commercial sources of dietary supplements. This study examined 38 samples, including voucher and commercial samples. All eight commercial candlenut dietary supplement samples were adulterated. Additionally, two samples sold as Brazil nuts were also found to be adulterated. Other nuts were screened for the presence of Cardiac Glycosides, but none were found to be positive. The presence of yellow oleander was confirmed in all commercial dietary supplement samples marketed as candlenut as well as in commercial samples of Brazil nut. This study provides simple key identification characters using micro-morphology and histochemical localization of cardiac glycosides in the commercial nuts, HPTLC fingerprints, and LC-DAD-Q-ToF analytical parameters to detect and identify adulteration in commercial products.
Assuntos
Bertholletia , Suplementos Nutricionais , Suplementos Nutricionais/análise , Bertholletia/química , Contaminação de Alimentos/análise , Cromatografia em Camada Fina , Nozes/química , Cromatografia Líquida de Alta Pressão , Redução de Peso , MicroscopiaRESUMO
Salvia mellifera, native to California, Baja California, and Mexico, is a medicinal herb traditionally used to relieve pain, body aches, including chronic pain. A detailed phytochemical investigation of aerial parts of S. mellifera was accomplished to find species-specific markers and to differentiate the closely related, often (un)intentionally substituted with S. apiana. A total of 22 metabolites, including flavonoids (1-14), triterpenoids (15-18), diterpenoids (19-21), and phenylpropanoid (22), were isolated and characterized thoroughly. Among the isolates, eupatorin 3'-O-glucopyranoside (1) was identified as undescribed phytochemical and detailed structure elucidation was achieved through extensive NMR and mass spectral data analysis.
Assuntos
Salvia , Salvia/química , Glucosídeos/análise , México , Flavonoides/química , Componentes Aéreos da Planta/química , Compostos Fitoquímicos/análiseRESUMO
ABSTRACT Fadogia agrestis Schweinf. ex Hiern (Vangueria agrestis (Schweinf. ex Hiern) Lantz), Rubiaceae, is an African traditional medicinal plant also used as a dietary supplement in the US. The present paper is the first report of the pharmacognostic study of the leaf, stem and root of F. agrestis by microscopy, HPTLC and total phenolic/flavonoid content analyses. Noteworthy microscopic features that can help in identification and quality control are septate and lignified non-glandular trichomes on leaf and stem epidermises, paracytic stomata on leaf abaxial epidermis, numerous cells containing yellow substances of presumably phenolic compounds in leaf and stem, calcium oxalate druses and prismatic crystals in leaf and styloids in stem, primary phloem fibers in stem, brachysclereids in stem and root, spherical starch grains in root, and vessels with vestured pits and simple perforated end walls. In addition to microscopy, a total phenolic/flavonoid content determination and an HPTLC method were also developed for rapid chemical fingerprint analyses of Fadogia samples and dietary supplements.
RESUMO
BACKGROUND: Medicinal plants are an important source to identify new active pharmaceutical compounds. Traditionally, the sap of Euphorbia umbellata is widely used to treat cancer and inflammatory conditions. These effects have been attributed to the presence of terpenes and phenolic compounds in the extracts of this plant. Euphol, a tetracyclic triterpene alcohol, is one of the major compounds present in Euphorbia species, and some biological activities have been attributed to this compound. PURPOSE: This study aimed to evaluate the in vitro cytotoxicity of euphol against Jurkat, HL-60, K-562, B16F10, and HRT-18 cells lines, as well as the biological stability, distribution, metabolism properties in vitro, and the determination of the concentration of euphol in the plasma and liver of rats. METHODS: The MTT reduction assay was used to evaluate the cytotoxicity of euphol against cancer cell lines, and the selectivity index, the morphology and cell cycle assays to evaluate the death mechanisms in K-562 and B16F10 lineages. UHPLC-MS was applied for the in vivo evaluation of the concentration of euphol in plasma and liver, and in vitro metabolic stability in human liver microsomes and S9 fraction, plasma protein binding, and stability in simulated gastric and intestinal fluids assays. CONCLUSIONS: This study demonstrated that euphol exhibited cytotoxic effects against a variety of cancer cells lines, selectivity against leukemia and possibly, the mechanism involved is apoptosis. The evaluation of stability, distribution, and metabolism properties showed that euphol was unstable in gastric and intestinal fluids, presenting moderate plasma protein binding with two hours elimination half-life and possible phase II liver metabolism. All the results suggested that further studies could be developed to prove the viability of euphol as an anticancer agent.
Assuntos
Euphorbia/química , Lanosterol/análogos & derivados , Látex/química , Animais , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Humanos , Células Jurkat , Lanosterol/farmacologia , Extratos Vegetais/farmacologia , Plantas Medicinais/química , RatosRESUMO
OBJECTIVES: The objective of this study was to evaluate the dental color exposed to acute cigarette smoke treatment and quantify the amount of nicotine in samples exposed to cigarette smoke, after dental prophylaxis and after in-office bleaching. MATERIALS AND METHODS: Sixty-nine healthy human molars were subjected to cigarette smoke in a cigarette machine. The teeth were divided into three groups: positive control, prophylaxis, and bleaching. Forty cycles of smoke exposition with duration of 15 min each were performed using 10 cigarettes (positive control). Dental prophylaxis was performed with a rotating brush and prophylaxis paste; in-office bleaching was performed with 35% hydrogen peroxide, in two sessions of three 15-min applications, with a 1-week interval between sessions. The color was evaluated at the baseline, after exposure to cigarette smoke, after dental prophylaxis, and after in-office bleaching. Teeth from each group were powdered and analyzed by gas chromatography-mass spectrometry in order to measure the amount of nicotine present in each group. Data from quantification of nicotine and color change were analyzed by one-way ANOVA and Tukey's test (α = 0.05). Data for subjective and objective color evaluation, a perceptible dental darkening occurred in teeth after exposure to cigarette smoke. Dental prophylaxis was able to recover the original color of teeth however, only after bleaching teeth became whiter than at the baseline (p < 0.001). The amount of nicotine was significantly different and higher in positive control group (3.3 ± 1.3 µg/g of tooth), followed by the prophylaxis group (2.1 ± 1.4 µg/g) and the bleaching group (0.8 ± 0.3 µg/g) (p < 0.001). CONCLUSIONS: Cigarette smoke penetrates into the dental structure. Dental prophylaxis and bleaching with 35% hydrogen peroxide can partially remove the nicotine from tobacco smoke. However, when in-office bleaching was applied, a more significant nicotine removal was achieved. CLINICAL SIGNIFICANCE: Dental prophylaxis could remove most of the external nicotine-staining on the tooth surfaces while bleaching could further reduce the external and internal nicotine-staining of teeth.
Assuntos
Profilaxia Dentária , Nicotina/análise , Fumar , Clareamento Dental , Descoloração de Dente/terapia , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Técnicas In Vitro , Dente MolarRESUMO
ABSTRACT Euphorbia umbellata (Pax) Bruyns, Euphorbiaceae, is commonly used in folk medicine of southern Brazil to treat several kinds of cancer. The latex (part of the plant used for this purpose) is mixed with water and taken as treatment; but this matrix contains toxic potential related to the presence of some phorbol type diterpenes. So the aim of this study was to evaluate the cytotoxicity of the crude extract of the bark of E. umbellata and its fractions (Hex, CHCl3, EtOAc and MeOH) using in vitro assay (applying Jurkat cells line). A preliminary cytotoxic study (MTT reduction, trypan blue exclusion and DNA quantification assays) was executed to identify the most active material. The CHCl3 fraction displayed the highest activity and was selected for further investigation of any cytotoxic mechanism and evaluation of chemical composition; flow cytometry, Acridine orange and Hoechst 33342 staining experiments and Gas chromatography–mass spectrometry analysis were applied to achieve these results. This fraction demonstrated the best cytotoxic results against Jurkat cells line with IC50 of 29.00 ± 1.49, 10.06 ± 1.48 and 4.83 ± 2.25 µg/ml for 24, 48 and 72 h of experiment, respectively (trypan blue exclusion). The mechanism responsible for this action can be associated with the promotion of cell cycle arrest and apoptosis. The two main classes of compounds present in the CHCl3 fraction are steroids and triterpenes. Further, phytochemical studies with this fraction need to be evaluated, to try isolating these substances and establishing a more detailed cytotoxic study against Jurkat cells.
RESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Euphorbia umbellata latex (sap) has normally been used in folk medicine in southern Brazil to treat different types of cancers. AIM OF STUDY: To carry out a biomonitored investigation of partitioned latex using in vitro assay, to identify the main mechanisms related with the action of the most active fraction as well as to develop a phytochemical study with this material. MATERIALS AND METHODS: Biological screening was performed with hexane, chloroform, ethyl acetate and methanol fractions from the latex of E. umbellata using MTT, trypan blue, and neutral red assays to determine the cytotoxicity against HRT-18, HeLa and Jurkat cells and flow cytometry, DNA quantification, acridine orange and Hoechst 33342 staining to investigate mechanisms of action for the hexane extract. The phytochemical study of the hexane fraction was performed by chromatographic procedures and the substances were identified by NMR analysis. The isolated terpenes were evaluated using MTT to determine the cytotoxicity against Jurkat cells. RESULTS: All the fractions presented concentration and time dependent cytotoxicity. The hexane fraction showed the highest cytotoxicity; whereas the Jurkat cell was the lineage with the highest sensitivity (IC50 1.87µg/mL). Fragmentation of DNA and apoptosis are two mechanisms related with the toxicity of hexane fraction. The hexane fraction arrested the cell cycle in the G0/G1 phase, and the selectivity index was 4.30. Phytochemical study of the hexane fraction led to isolation of euphol (main compound) and germanicol acetate. Both substances demonstrated some slight cytotoxic activity against Jurkat cells after 72h; however the activity was minimal compared to vincristine (anticancer standard drug). CONCLUSION: The current research proves that the fractions of the latex from E. umbellata have a cytotoxic effect against three different cancer cells lines. The hexane fraction showed high in vitro cytotoxic effects against Jurkat cells demonstrating that the effect may be due to non-polar constituents. The two isolated terpenes (euphol and germanicol acetate) showed poor cytotoxic activity indicating that the anticancer properties of the extract may be caused by other substances present in the hexane fraction.
Assuntos
Citotoxinas/farmacologia , Euphorbia/química , Extratos Vegetais/farmacologia , Apoptose/efeitos dos fármacos , Brasil , Linhagem Celular Tumoral , Citotoxinas/química , Fragmentação do DNA/efeitos dos fármacos , Fase G1/efeitos dos fármacos , Células HeLa , Humanos , Células Jurkat , Medicina Tradicional/métodos , Compostos Fitoquímicos/química , Compostos Fitoquímicos/farmacologia , Extratos Vegetais/química , Fase de Repouso do Ciclo Celular/efeitos dos fármacosRESUMO
BACKGROUND: Maca is an Andean crop of the Brassicaceae family which is mainly known for its fertility-enhancing properties following consumption. The hypocotyls display various colours ranging from white to black. Each colour has different biological effects. The aim of this study was to analyse the concentrations of major secondary metabolites in hypocotyls and leaves of maca in a controlled planting experiment in the Peruvian Andes at 4130 m above sea level. The effects of colour type and of previous cultivation of the field were examined. RESULTS: In the hypocotyls, the colour type effect was significant for most secondary metabolites; exceptions were beta-sitosterol and campesterol. The lead-coloured, yellow and violet maca hypocotyls were rich in glucosinolates, macaene and macamides, respectively. Previous cultivation affected macaene, campesterol and indole glucosinolate concentrations. Effects on metabolite concentrations in the leaves were minor. Hypocotyls were richer in macaene, macamides and glucosinolates than were leaves, and were poorer in beta-sitosterol and total phenols. CONCLUSION: Colour type has to be considered in maca production, as colour associates with variations in concentrations of distinct bioactive metabolites. Leaves may be interesting for animal nutrition purposes as they contain essentially the same secondary metabolites as the hypocotyls but in clearly lower concentrations.