The aim of this study was to determine the chemical variability of Allamanda cathartica flowers collected in different localities of Santa Catarina State, Brazil. Extractive solutions (25 samples) were analyzed by HPLC-DAD for the qualitative and quantitative profiles of the iridoid plumieride, the flavonoid rutin as well as total flavonoid content expressed in rutin. The chemical variability was evaluated by Principal Component Analysis (PCA) and Hierarchical Clustering Analysis (HCA). The samples demonstrated similar chromatographic profiles. In PCA analysis, Araquari samples AC01, AC08 and AC25 did not group with other samples obtained from other locations. In the HCA analysis, it was possible to observe hierarchially similar groups, generating 5 clusters. Group 5 (samples AC01, AC08 and AC25) presented lower similarity compared to the other groups. This dissimilarity can be explained by the amount of metabolites from each collection. Regarding the content of markers, it was observed a variation between 27 to 230 mg/g, 2.7 to 9.5 mg/g and 7.14 to 17.8 mg/g for plumieride, rutin, and total flavonoids, respectively. The samples from Araquari (AC01, AC08, AC25) presented the highest concentrations of chemical markers. Therefore, for reproducibility and standardization of A. cathartica flower extracts, it is important to determine the location and the plant cultivation conditions.

Abstract Solvents play important and critical role in natural product chemistry and could generate artefacts during the extraction and purification of metabolites from a biological matrix. This study aimed to correlate the chromatographic profile with biological activity of Ipomoea pes-caprae (L.) R. Br., Convolvulaceae, extracts obtained with hydroethanolic extraction. Thus, aerial parts of I. pes-caprae were extracted with different concentration of ethanol (50, 70 and 90°GL) and the obtained extracts were analysed by HPLC–UV. HPLC data were studied employing chemometrics to discriminate the samples. Moreover these samples were further characterized by using UPLC–QTOF/MS data. The extracts were also biomonitored through the paw-oedema and spontaneous nociception induced by trypsin in mice. Different chromatographic profiles were obtained and the exploratory analysis clearly revealed higher level of ethyl caffeate in extracts of lower strength of ethanol (50°GL). This compound was suggested to be an artefact formed by transesterification of caffeoylquinic acid derivatives present in the plant, once it was not observed when other solvents were employed. During the biological assay, only the extract obtained with ethanol 50°GL presented significant inhibition of inflammation (45 ± 9%) and nociception (24 ± 3%). Ethyl caffeate seems to be linked to the anti-inflammatory effect since it reduced 86 ± 5% of paw-oedema induced by trypsin. Artefacts could contribute to the biological activity of herbal preparations and consequently lead to misinterpretation of the results.

A simple and efficient microwave-assisted extraction process has been developed in order to extract plumieride from Allamanda cathartica flowers. The extracts have been obtained through microwave-assisted extraction (MAE) process with ethanol and ethyl acetate at different time, power and temperatures. Static maceration at room temperature with ethanol and ethyl acetate was obtained in order to compare the efficiency of extraction. HPLC method was validated to quantify plumieride in the extracts. The analyzed parameters: linearity (R2 = 0.999), precision (CV 3.3%), recovery (98%), specificity (97%), limit of detection (1 µg mL-1), and limit of quantification (2 µg mL-1) were satisfactory. The yield of plumieride reached 43% in the extracts under the optimal MAE conditions (10 min, 300W). Plumieride recoveries by conventional extraction method were 12% with ethanol and 22% with ethyl acetate. Time and power parameters on plumieride extraction yield was determined by applying a factorial design and surface plots. Ethanol was the best extractor solvent and it was possible to obtain high contents (52 mg g-1 of dry flowers) at optimized conditions. It can be concluded that MAE process has actual advantages over the conventional extraction method in terms of shorter time and higher efficiency to recover plumieride from A. cathartica flowers.

Abstract The essential oil of Chenopodium ambrosioides L., Amaranthaceae, was obtained by steam distillation in a Clevenger apparatus and characterization was performed using chromatographic and spectroscopic assays (GC-FID, GC/MS, 1H NMR). Two major compounds were identified: p-cymene (42.32%) and ascaridole (49.77%). The ethanolic extract and hydrolate were fractionated by liquid–liquid partitioning and the compounds were characterized by GC/MS. The essential oil, ethanol extract and fractions by partitioning with dicloromethane, ethyl acetate and butanol were tested in tumor cell lines (K562, NALM6, B15, and RAJI). Significant cytotoxic activity was found for essential oil (IC50 = 1.0 µg/ml) for RAJI cells and fraction dicloromethane (IC50 = 34.0 µg/ml) and ethanol extract (IC50 = 47.0 µg/ml) for K562 cells. The activity of the essential oil of C. ambrosioides is probably related to the large amount of ascaridol, since the other major compound, p-cymene, is recognized as a potent anti-inflammatory and has low cytotoxic activity.

The aim of this research was to evaluate the fractions obtained from the leaf, stem and roots of Allamanda schottii Pohl, Apocynaceae, responsible for the cytotoxicity, using several cell lines. Cytotoxicity was correlated with the season the part of the plant, and the major compounds were assessed. The ethanol extracts of leaves, stems and roots obtained at different seasons were evaluated in the human erythromyeloblastoid leukemia cell line (K562). Subsequently the ethanol extracts and dichloromethane fractions collected in winter were evaluated in mouse fibroblast cell line (Mus musculus) (L929), cervix adenocarcinoma (HeLa), human pre-B leukemia (Nalm6), as well as K562 cell line. The compounds plumericin, plumieride and ursolic acid isolated from ethanol extracts of the stems were evaluated in the same cell lines, as well as on breast adenocarcinoma cell line (MCF-7), and Mus musculus skin melanoma cell line (B16F10). The chromatographic profiles of the dichloromethane fractions were obtained by high performance liquid chromatography. The results revealed that the season during which A. schottii was collected, and the part of the plant analyzed, influence the cytotoxicity on the K562 cells tested. On the other hand the dichloromethane fractions, mainly from the stems and roots, are responsible for the cytoxicity on the cells tested. These results may be associated with the seasonal variation of plumericin in these parts of the plant. This information is in accordance with the HPLC analysis. The results clearly show the potential for the phytotherapeutic use of this species, and suggest that the cytotoxic activity observed may be due to the presence of plumericin, or to minor compounds not yet identified. The seasonal influence on the production of secondary metabolites was verified.

This paper evaluates CHCl3 and CH3OH extracts of the stem bark, branches and leaves of Drimys brasiliensis and drimane sesquiterpenes isolated from the stem bark against strains of Leishmania amazonensis and Leishmania braziliensis promastigotes and Plasmodium falciparum trophozoites. All of the extracts and compounds were tested in cell lines in comparison with reference standards and cell viability was determined by the XTT method. The CHCl3 and CH3OH extracts from the stem bark and branches yielded promising results against two strains of Leishmania, with 50% inhibitory concentrations (IC50) values ranging from 39-100 µg/mL. The CHCl3 extract of the stem bark returned IC50 values of 39 and 40.6 µg/mL for L. amazonensis and L. braziliensis, respectively. The drimanes were relatively effective: 1-β-(p-coumaroyloxy)-polygodial produced IC50 values of 5.55 and 2.52 µM for L. amazonensis and L. braziliensis, respectively, compared with 1-β-(p-methoxycinnamoyl)-polygodial, which produced respective IC50 values of 15.85 and 17.80 µM. The CHCl3 extract demonstrated activity (IC50 of 3.0 µg/mL) against P. falciparum. The IC50 values of 1-β-(p-cumaroyloxyl)-polygodial and 1-β-(p-methoxycinnamoyl)-polygodial were 1.01 and 4.87 µM, respectively, for the trophozoite strain. Therefore, the results suggest that D. brasiliensis is a promising plant from which to obtain new and effective antiparasitic agents.

Três triterpenos pentacíclicos inéditos foram isolados da resina de Protium kleinii. Suas estruturas, 3-oxo-11beta,16beta-diidróxi-urs-12-eno (1), 3-oxo-11beta-hidróxi-urs-12-eno (2) e 3-oxo-11beta-hidróxi-olean-12-eno (3), foram elucidadas por espectroscopia de RMN. A administração sistêmica destes compostos (10-100 mg kg-1, i.p.) foi capaz de inibir tanto a primeira quanto a segunda fase da nocicepção induzida pela formalina em camundongos. Em adição, foram isolados os triterpenos conhecidos 3beta,16beta-dihidróxi-urs-12-eno (breina) (4) e uma mistura de alfa (5)- e beta-amirina (6).

Three novel pentacyclic triterpenes have been isolated from the resin of Protium kleinii and their structures elucidated by NMR spectroscopy as 3-oxo-11beta,16beta-dihydroxy-urs-12-ene (1), 3-oxo-11beta-hydroxy-urs-12-ene (2) and 3-oxo-11beta-hydroxy-olean-12-ene (3). The systemic administration of the above compounds (10-100 mg kg-1, i.p.) was able to inhibited both first and second phase of the formalin-induced nociception in mice. The known triterpenes 3beta,16beta-dihydroxy-olean-12-ene (brein) (4) and a mixture of alpha (5)- and beta-amyrin (6) were also found.

There are NMR data of ¹H and 13C of the iridoid plumieride, but controversy related to the assignments of the protons H-3 or H-10 and carbons C-6 or C-7 and C-3 or C-10 are described in the literature. There are a little discussion regarding to the resonance assignment of protons of the glycoside unity. Analysis based on 2D shift correlated NMR spectra (COSY, HETCOR, HETCORLR) and NOE difference ¹H NMR spectra allowed to assign unambigously the chemical shift of ¹H and 13C of plumieride which has been found in the literature with non coincident values.