RESUMO
INTRODUCTION: In Brazil, the plant group popularly known as "pedra-ume-caá" is used in folk medicine for the treatment of diabetes, and its raw material is commonly sold. OBJECTIVE: The aim of the study was to apply a method for chemical identification of extracts of dry pedra-ume-caá leaves using HPLC-high-resolution mass spectrometry (HRMS) and NMR and develop a multivariate model with NMR data to authenticate commercial samples. In addition, to evaluate the biological activities of the extracts. MATERIALS AND METHODS: Dry extracts of Myrcia multiflora, Myrcia amazonica, Myrcia guianensis, Myrcia sylvatica, Eugenia punicifolia leaves, and 15 commercial samples (sold in Manaus and Belém, Brazil) were prepared by infusion. All the extracts were analysed using HPLC-high-resolution mass spectrometry (HRMS), NMR, principal component analysis (PCA), and hierarchical cluster analysis (HCA). The antidiabetic effect of extracts was evaluated according to enzymatic inhibition. Their content of total phenols, cell viability, and antioxidant and antiglycation activities were also determined. RESULTS: HPLC-HRMS and NMR analysis of these extracts permitted the identification of 17 compounds. 1H NMR data combined with multivariate analyses allowed us to conclude that catechin, myricitrin, quercitrin, and gallic and quinic acids are the main chemical markers of pedra-ume-caá species. These markers were identified in 15 commercial samples of pedra-ume-caá. Additionally, only the extracts of M. multiflora and E. punicifolia inhibited α-glucosidase. All the extracts inhibited the formation of advanced glycation end products (AGEs) and showed free-radical-scavenging activity. These extracts did not present cytotoxicity. CONCLUSION: This study revealed the chemical markers of matrices, and it was possible to differentiate the materials marketed as pedra-ume-caá. Moreover, this study corroborates the potential of these species for treating diabetes.
Assuntos
Diabetes Mellitus , Myrtaceae , Antioxidantes/química , Extratos Vegetais/química , Myrtaceae/química , Espectroscopia de Ressonância Magnética , Folhas de Planta/químicaRESUMO
The genus Cecropia is used in the traditional medicine of Tabasco, Mexico, in diabetes and hypertension treatments, mainly without distinction of the species. This contribution aimed to carry out the metabolic analysis and Proton Nuclear Magnetic Resonance (1H-NMR) spectroscopy-based fingerprinting of the hydroalcoholic leaf extracts of Cecropia peltata (Cp) and Cecropia obtusifolia (Co) collected in five sub-regions of the State of Tabasco (Cp1, "Centro"; Cp2, "Chontalpa"; Cp3, "Pantanos"; Cp4, "Ríos" and Co5, "Sierra"). Firstly, the extracts were evaluated for their Total Phenol Content (TPC) and Total Flavonoid Content (TFC) by spectrophotometric methods. In addition, metabolic analysis was performed using High-Performance Liquid Chromatography with Diode-Array Detection HPLC-DAD, which allowed the quantification of the chemical markers: chlorogenic acid, isoorientin, and orientin, as well as a vitexin analog. Finally, metabolomic analysis was carried out based on the 1H-NMR spectra. The Cp4 extract (C. peltata from the "Ríos" sub-region) presented the highest values of TPC (155 ± 9.1 mg GAE/g E) and TFC (724 ± 22.2 mg RE/g E). The metabolic analysis was similar among the five samples; the highest concentrations of the four chemical markers were found in Cp3 (C. peltata from the "Pantanos" sub-region) for chlorogenic acid (39.8 ± 2.3 mg/g) and isoorientin (51.5 ± 2.9 mg/g), in Cp4 for orientin (49.9 ± 0.6 mg/g), and in Cp2 (C. peltata from the "Chontalpa" sub-region) for the vitexin analog (6.2 ± 0.2 mg/g). The metabolic analysis and the 1H-NMR fingerprint analysis showed intraspecies differences among the C. peltata samples and interspecies between C. peltata and C. obtusifolia, which were attributed to variations in the metabolite groups as well as in the proportion of sugars such as glucose and xylose.
RESUMO
This study investigated the similarities between Echinodorus macrophyllus and Echinodorus grandiflorus, plant species that are traditionally used in Brazil to treat rheumatism and arthritis, whose anti-inflammatory effects are supported by scientific evidence. The contents of cis- and trans-aconitic acid, homoorientin, chicoric acid, swertisin, caffeoyl-feruloyl-tartaric acid, and di-feruloyl-tartaric acid were quantified by UPLC-DAD in various hydroethanolic extracts from the leaves, whereas their anti-oxidant activity and their effect on TNF release by LPS-stimulated THP-1 cells were assessed to evaluate potential anti-inflammatory effects. The 50% and 70% ethanol extracts showed higher concentrations of the analyzed markers in two commercial samples and a cultivated specimen of E. macrophyllus, as well as in a commercial lot of E. grandiflorus. However, distinguishing between the species based on marker concentrations was not feasible. The 50% and 70% ethanol extracts also exhibited higher biological activity, yet they did not allow differentiation between the species, indicating similar chemical composition and biological effects. Principal component analysis highlighted comparable chemical composition and biological activity among the commercial samples of E. macrophyllus, while successfully distinguishing the cultivated specimen from the commercial lots. In summary, no differences were observed between the two species in terms of the evaluated chemical markers and biological activities.
RESUMO
Trace elements bioaccumulation patterns can be an important tool to assess differences among cetaceans' populations. In this work, their use as potential chemical markers to differentiate franciscanas (Pontoporia blainvillei) populations was evaluated. Franciscanas were collected from three states in southeastern Brazil, which comprise three different Franciscana Management Areas (FMAs): Espírito Santo (FMA Ia), southern Rio de Janeiro (FMA IIa), and central São Paulo (FMA IIb). The concentrations of As, Cd, Cu, Fe, Hg, Mn and Zn were determined in the muscle, liver and kidney of the animals. Cadmium was the most valuable chemical marker to differentiate stocks, separating at least FMA IIa from the others. The higher Cd levels in FMA IIa, along with dietary information, indicate that the predominant consumption of cephalopods by this population is the main reason for the differences found. Additionally, environmental characteristics of the areas should also be considered as divergent sources of trace elements. Our findings suggest that non-essential trace elements, such as Cd, can be successful markers to differentiate populations. The Mn concentrations in FMA Ia raised concern and must be carefully monitored, as well as other elements that compose the iron ore tailings that have impacted the Espírito Santo coastal area. Additionally, this is the first study to report trace element concentration in the franciscanas from FMA IIa (southern Rio de Janeiro). Trace element concentrations found in franciscanas may represent different contamination levels in their preys and environments, which might pose specific threats to distinct populations. Therefore, our findings are important to characterize and differentiate franciscana populations and to guide precise management and conservation actions for the distinct stocks of this endangered species.
Assuntos
Golfinhos , Oligoelementos , Poluentes Químicos da Água , Animais , Oceano Atlântico , Bioacumulação , Brasil , Cádmio , Monitoramento Ambiental , Oligoelementos/análise , Poluentes Químicos da Água/análiseRESUMO
Abstract The Brazilian native species Cestrum intermedium, known as mata-boi, induces hepatotoxicity and death when ingested by cattle. While most studies on this species focus on toxicological features, our study is the first to describe the anatomy and in vitro biological activities of Cestrum intermedium. We investigated adult leaves and stems by histochemistry, described their anatomy, performed physical-chemical analysis, determined in vitro antioxidant and antimicrobial activities, and identified secondary metabolites. A few noteworthy anatomical features were the anomocytic stomata on the abaxial surface and the absence of trichomes, in addition to the circular shaped petiole with two projections on the adaxial surface. Histochemical analysis showed chemical markers such as alkaloids, usually reported as toxic, and terpenoids. Potassium nitrate (ATR-FTIR) and lupeol palmitate (NMR) were detected on the crude stem extract. Thermogravimetric and physical-chemical analysis provided fingerprint parameters for the species. Minimal Inhibitory Concentration (MIC) assay revealed that Staphylococcus aureus, Staphylococcus epidermidis, and Candida albicans were weakly inhibited by extract samples. Chloroform and ethyl acetate fractions presented high phenolic content, which resulted in in vitro antioxidant activity. These novel features expand the knowledge about this species, considering that previous studies mainly focused on its toxicity. Our study also provided characteristics that may help in avoiding misidentification between Cestrum members, especially when taxonomic keys cannot be employed, as in the absence of flowers and fruits.
Assuntos
Técnicas In Vitro/métodos , Solanaceae/anatomia & histologia , Solanaceae/classificação , Staphylococcus epidermidis , Terpenos/efeitos adversos , Microscopia Eletrônica de Varredura/métodos , Caules de Planta/anatomia & histologia , Folhas de Planta/anatomia & histologiaRESUMO
The cocoa bean shell (CBS) is one of the main cocoa byproducts with a prospective to be used as a functional food ingredient due to its nutritional and sensory properties. This study aims to define the chemical fingerprint of CBSs obtained from cocoa beans of diverse cultivars and collected in different geographical areas of Venezuela assessed using high-performance liquid chromatography coupled to photodiodes array and mass spectrometry (HPLC-PDA-MS/MS) and spectrophotometric assays combined with multivariate analysis for classification purposes. The study provides a comprehensive fingerprint and quantitative data for 39 compounds, including methylxanthines and several polyphenols, such as flavan-3-ols, procyanidins, and N-phenylpropenoyl amino acids. Several key cocoa markers, such as theobromine, epicatechin, quercetin-3-O-glucoside, procyanidin_A pentoside_3, and N-coumaroyl-l-aspartate_2, were found suitable for the classification of CBS according to their cultivar and origin. Despite the screening methods required a previous purification of the sample, both methodologies appear to be suitable for the classification of CBS with a high correlation between datasets. Finally, preliminary findings on the identification of potential contributors for the radical scavenging activity of CBS were also accomplished to support the valorization of this byproduct as a bioactive ingredient in the production of functional foods.
RESUMO
The application of 1H NMR spectroscopy and chemometrics for the analysis of extracts of Lantana camara is described. This approach allowed to predict the leishmanicidal activity of samples obtained at different harvest times from their 1H NMR spectra. The anti-leishmanial activity of dichloromethane extracts obtained from the aerial parts of L. camara was measured using an in vitro assay. As the extracts displayed differences in their activity according to a one-way ANOVA analysis, their 1H NMR spectra were subjected to multivariate analysis using exploratory (Principal Component Analysis (PCA) and Anova Simultaneous Component Analysis (ASCA)) and regression, (Partial Least Squares Regression to Latent Structures (PLS)) chemometrics methods. These analyses allowed to establish and characterize a predictive model capable of determining the anti-leishmanial activity of Lantana camara dichloromethane extracts from their 1H NMR spectra. Figures of merit of the developed method are given as well. The identified chemical signals responsible for the iPLS calibration model corresponded to the presence of eicosane, caryophyllene oxide, ß-ionone, tiglic acid, lantanilic acid, camaric acid, and lantadene B; the chemical markers. This study proposed a fast and simple method that avoids the need of using complex biological assays to predict the leishmanicidal activity of L. camara dichloromethane extracts.
Assuntos
Lantana , Leishmania , Cloreto de Metileno , Extratos Vegetais/farmacologia , Espectroscopia de Prótons por Ressonância MagnéticaRESUMO
The vasorelaxing effect of the methanol extract of the flowers of Crataegus gracilior, a Mexican medicinal plant used to treat some cardiovascular diseases, was assessed, and its possible chemical markers identified. The extract produced a potent vasodilator effect on isolated rat aortic rings (EC50 = 1.83 ± 1.39 µg/mL; Emax = 100 ± 3.4%). Vitexin, the most commonly identified flavonoid in the flowers and used to standardise some Crataegus species, was not found at all in this plant sample. Instead, daucosterol, and corosolic and euscapic acids were purified. The two triterpene acids have been reported to possess beneficial effects on cardiovascular diseases. These results indicate that the vasodilator effect might induce the hypotensive effect claimed by users, and that euscapic and corosolic acids may be the main vasodilator compounds, and can then be employed as the chemical markers towards the future standardisation of the extract.
Assuntos
Crataegus/química , Flores/química , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Vasodilatadores/farmacologia , Animais , Aorta/efeitos dos fármacos , Apigenina/análise , Relação Dose-Resposta a Droga , Flavonoides/análise , Masculino , Metanol , México , Plantas Medicinais/química , Ratos Wistar , Triterpenos/análise , Triterpenos/farmacologia , Vasodilatadores/análise , Vasodilatadores/químicaRESUMO
The essential oil of Psidium guajava L. has been studied for pharmacological and industrial purposes, without considering the plant's genotype regarding the heterogeneity of its composition. The present study aimed to characterize the chemotype diversity of the essential oil extracted from the leaves of 22 genotypes of P. guajava grown in two different environments in the state of Espírito Santo, Brazil, and to identify the different chemical markers present in these plants. Essential oil from the leaves of the P. guajava genotypes was extracted by hydrodistillation, and its chemical composition was analyzed by gas chromatography-flame ionization detection (GC-FID) and gas chromatography-mass spectrometry (GC-MS). Thirty-three compounds were identified, comprising 87.5-99.0% of the total composition, with a prevalence of sesquiterpenes in all samples. The major compounds identified consisted of (E)-trans-Caryophyllene, α-Humulene, trans-Nerolidol, ß-Bisabolene, ß-Bisabolol, and Hinesol, the first of which was identified as a possible chemical marker for the species. Multivariate factor analysis of the chemical composition of P. guajava oil identified three chemotypes: Commercial - PAL, SEC, PS, PET, C7, C11, and C17MI, characterized by high levels of ß-Selinene, α-Selinene, Hinesol, and 14-hydroxy-epi-(E)-caryophyllene, with ß-Selinene and α-Selinene as the chemical markers; C10 and C13, exhibiting high levels of Elemol, trans-Nerolidol, trans-ß-Eudesmol, and (2Z, 6Z)-Farnesol, which were indicated as chemical markers, and Cortibel - C1, C2, C3, C4, C5, C6, C8, C9, C12, C14, C15, C16, C17LI, which retained high levels of α-Cedrene, cis-α-Bergamotene, α-Humulene, Humulene epoxide, epi-α-Cadinol, ß-Bisabolol, and α-Bisabolol, with ß-Bisabolol and α-Bisabolol as the chemical markers. The use of guava genotypes with different chemotypes, that are agronomically favorable to fruit production and essential oil exploitation adds value to the crop and renders it more sustainable. Given guava crops produce large amounts of leaf biomass, resulting from successive prunings, the extraction of their essential oil, which retains commercially valuable compounds, can be feasible.
Assuntos
Óleos Voláteis/isolamento & purificação , Compostos Fitoquímicos/isolamento & purificação , Folhas de Planta/química , Psidium/química , Genótipo , Óleos Voláteis/química , Compostos Fitoquímicos/química , Psidium/genéticaRESUMO
An analytical method utilizing liquid chromatography coupled to mass spectrometry with electrospray ionization has been developed for the identification of prostaglandins (PGs) in cerebral tissues. The five compounds identified (thromboxane B2, prostaglandin E2, prostaglandin D2, 6-keto-prostaglandin F1 alpha and prostaglandin F2 alpha) are cellular mediators of inflammation and are involved in a variety of physiological and pathological processes by acting on membrane receptors on the surfaces of target cells. The parameters of the electrospray ionization interface were optimized to obtain the highest possible sensitivity for all compounds studied. The limits of detection ranged from 0.25 to 1.09⯵gâ¯L-1, and the limits of quantification ranged from 0.83 to 3.64⯵gâ¯L-1. The method was validated and applied to samples of brain tissue from five mice. The sample concentrations of the four prostaglandins quantified ranged from 375 ȵg L-1for prostaglandin E2 to 6602⯵gâ¯L-1 for prostaglandin D2. An advantage of this work that should be emphasized is the fast response of the method, which allows to obtaining the lipid profile after a 3â¯min chromatographic run.
Assuntos
Cérebro/química , Prostaglandinas/análise , Animais , Cromatografia Líquida de Alta Pressão , Camundongos , Estrutura Molecular , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em TandemRESUMO
Spondias tuberosa is a medicinal plant used by several local communities in northeast Brazil to treat infections, digestive disorders and inflammatory conditions. The study aimed to identify and quantify the major phenolic in hydroethanolic extract of leaves from S. tuberosa and to evaluate its anti-inflammatory potential. The chemical profile of extract was analyzed by HPLC-DAD and HPLC-MS. The in vivo anti-inflammatory activity was investigated in carrageenan-induced hind paw edema and peritonitis models in mice. Identified and quantified through HPLC-DAD or HPLC-MS analyses of S. tuberosa extract were the following compounds: chlorogenic acid, caffeic acid, rutin and isoquercitrin. The inflammatory response to carrageenan was significantly reduced in both models by S. tuberosa extract. In hind paw edema, the edematogenic response was reduced by up to 63.6% and the myeloperoxidase activity was completely inhibited. In the peritonitis model, the total cell migration into the peritoneal cavity was reduced by up to 65%. The results obtained give evidence of the anti-inflammatory action of S. tuberosa and suggest the potential therapeutic benefit of this plant on inflammatory conditions. The chlorogenic acid, caffeic acid, rutin and isoquercitrin identified and quantified in S. tuberosa leaves enable us to suggest that these compounds could be used as chemical markers for quality control of derivative products from this species. Copyright © 2016 John Wiley & Sons, Ltd.
Assuntos
Anacardiaceae/química , Anti-Inflamatórios/farmacologia , Cromatografia Líquida de Alta Pressão/métodos , Folhas de Planta/química , Espectrometria de Massas em Tandem/métodos , Animais , CamundongosRESUMO
Wild adult specimens of the Peruvian anchovy Engraulis ringens were captured and reared to validate the daily periodicity of otolith microincrement formation. The postcapture stress generated spontaneous spawning, making it possible to conduct a rearing trial on larvae first in an artificial nutrient-enriched system (ANES) for 52 days followed by an artificial feeding regime in a culture tank until day 115 post-hatch. Microincrements of the sagittal otoliths of sacrificed juveniles [mean ± S.D. total length (LT ) = 5·13 ± 0·37 cm, range 5-6 cm; c.v. = 7·5%] showed very distinct light and dark zones. The slope of the relationship between the total number of increments after the hatch check and days elapsed after hatching was not significantly different from 1. The transfer from ANES to the artificial feeding regime induced a mark in the sagittal otoliths. The number of microincrements after this induced mark coincided with the number of days elapsed after the transfer date. In parallel experiments, adult E. ringens (mean ± S.D. LT = 14·92 ± 0·55 cm, range 13-16 cm) were exposed to one of two fluorescent marking immersion treatments with either alizarin red S (ARS; 25 mg l(-1) per 6 h) or oxytetracycline hydrochloride (OTC; 200 mg l(-1) per 10 h). The microincrements between fluorescent bands were distinct, ranging from 0·89 to 2·75 µm (mean ± S.D. =1·43 ± 0·28 µm; c.v. = 32%) and from 0·71 to 2·89 µm (1·53 ± 0·27 µm; c.v. = 35%) for ARS and OTC, respectively. The relationship between the number of microincrements between marks and the number of elapsed days for ARS and OCT treatments indicated that there was a significant correspondence between the number of increases observed and the number of days. Hence, daily microincrements of otoliths of E. ringens are likely to be formed in juveniles and adults under natural conditions.
Assuntos
Peixes/crescimento & desenvolvimento , Membrana dos Otólitos/crescimento & desenvolvimento , Animais , PeriodicidadeRESUMO
The antioxidant properties and the foliar phenol composition of 25 Mexican varieties of Phaseolus vulgaris L. (common bean) were evaluated. Phaseolus coccineus was analysed with comparative aims. The high performance liquid chromatography with photodiode array detection analysis revealed 27 phenolics in the leaves of P. vulgaris (13 quercetin-3-O-glycosides, 8 kaempferol-3-O-glycosides, 2 myricetin glycosides and 4 phenolic acids) and 5 in P. coccineus (2 kaempferol-3-O-glycoside, 2 apigenin-7-O-glycoside and 1 luteolin-7-O-glycoside). All extracts showed high levels of phenols and flavonoids (0.964-5.601 mg g⻹ dry tissue, and 0.287-1.418 mg g⻹ dry tissue, respectively) and relevant antioxidant properties, suggesting that the leaves of the varieties of P. vulgaris are a significant source of natural antioxidants. The foliar phenol profiles were species-specific and, besides, the qualitative variation allowed discriminating among varieties of P. vulgaris. These profiles can represent an important varietal authenticity proof.