Resumo em Inglês:ABSTRACT “Sasam (沙參)” is a crude drug that is defined in the in Korean Herbal Pharmacopoeia as the root of Adenophora triphylla var. japonica (Regel.) Hara or A. stricta Miq., Campanulaceae. The dried roots of the Adenophora spp. are available in markets, and the roots of various species are similar to each other in shape, making it difficult to distinguish one from another using only the outer morphological appearance. Therefore, the present study aimed to establish quality control parameters for pharmacognostic evaluation and differentiation of five Adenophora species and two varieties grown in Korea. Inner morphological evaluation of the root of these plants was accomplished and preliminary chemical analysis was performed by liquid chromatography-mass spectrometry profiling. As a result, significant differences among samples were found in anatomical characteristics such as number and thickness of cork layer, existence of stone cell in cork layer, frequency of vessels, and area of intercellular space. Significant differences were found among the samples in the content of three components including shashenoside I and a new alkyl glycoside, adenophoroside I. These findings could provide the scientific criteria for the proper identification and establishment of standards for the use of “Sasam”.
Resumo em Inglês:ABSTRACT Macamides and Macaenes are the bioactive marker compounds in maca (Lepidium meyenii Walp., Brassicaceae) tuber. To simultaneously quantify these two types of compound, HPLC method was studied. To distinguish and group the growing regions of different maca samples, Hierarchical cluster analysis, a chemometric method, was applied to analyze the HPLC data. The calibration curves obtained using the HPLC method showed satisfactory linearity with determination coefficients >0.9998. The precision and repeatability relative standard deviation values were <4%, and the accuracy relative standard deviation value was <5%. The limits of detection was <0.1 µg/ml and the limit of quantification was <0.3 µg/ml. Our HPLC method was successfully used for the separation and determination of macamides and macaenes in Maca within 45 min, i.e., two macaenes (9-oxo-10E,12Z-octadecadienoic acid and 9-oxo-10E,12E-octadecadienoic acid) and five macamides (N-benzyl-9-oxo-10E,12Z-octadecadienamide, N-benzyl-9-oxo-10E,12E-octadecadienamide, N-benzyl-9Z,12Z,15Z-octadecatrienamide, N-benzyl-9Z,12Z-octadecadienamide and N-benzyl-hexadecanamide). The HPLC method was applied to analyze and quantify the seven compounds in thirty maca samples with different colors and origins. The origins of all the maca samples were distinguished and grouped using hierarchical cluster analysis of the HPLC data. Accordingly, the metabolism of macaenes and macamides in maca post-harvest processing has also been proposed. The HPLC method is efficient to simultaneously quantify the macamides and macaenes in maca. Analyzing the HPLC data using hierarchical cluster analysis can distinguish maca growing origins.
Resumo em Inglês:ABSTRACT Citrus fruits are recognized as an important source of bioactive molecules such as limonin and nomilin. However, these molecules exhibit low bioavailability, therefore, obtaining these molecules using biotechnological techniques may be an alternative to harvesting them directly from fruits. The aim of this study was to quantify and identify limonoids in the dichloromethane extracts of Citrus seeds of Criolla orange, Oneco tangerine, Tangerine-lemon, Sour orange and Valencia orange from department of Antioquia-Colombia by high performance liquid chromatography with diode array detection, and high-resolution mass spectrometry. Although in all the samples total glycosidic free limonoids were present, Oneco tangerine seeds had the highest concentration, followed by Tangerine-lemon seeds, equivalent to 0.75% and 0.53% per total dry weight, respectively. These results suggest Oneco tangerine seeds may be used as an elite material for biotechnological processes looking for increased production of limonoids to support research and drug development.
Resumo em Inglês:ABSTRACT Chemical investigation of the aqueous fraction of the ethanol extract from the Brazilian endemic marine sponge Clathria (Clathria) nicoleae Vieira de Barros, Santos & Pinheiro, 2013, Microcionidae, sampled from a 55 m deep rhodolith bed at the Amazon River mouth, led to the isolation of a new hexapeptide, clathriamide (1). HP-20 resin was used to capture compound 1 from the aqueous fraction, which was purified by additional chromatographic steps. The absolute configuration of the amino acids of 1 was determined by advanced Marfey's analysis using 5-fluoro-2,4-dinitrophenyl-Nα-L-tryptophanamide. The amino acid derivatives analyzed by ultra-performance liquid chromatography coupled to a mass spectrometry using a C8 column enabled a good chromatographic resolution of L-Ile and L-allo-Ile, previously unfeasible using C18 column.
Resumo em Inglês:ABSTRACT The excessive use of chemical insecticides has led to negative effects on human health and the environment. Volatile oils are one of the possible potential alternatives to chemical insecticides. Traditionally Seriphidium brevifolium (Wall. ex DC.) Ling & Y.R.Ling, Asteraceae, powder from its leaves is used to treat gastric problems and expel intestinal worms by local peoples, but yet there is no literature available regarding its insecticidal activity. In this study fumigant toxicity and enzyme inhibition activities of the S. brevifolium volatile oil collected from the highlands of Skardu Baltistan, Pakistan, was evaluated against the red imported fire ant Solenopsis invicta. The phytochemical studies indicated that monoterpenes were the most abundant constituents, accounting for 88% of the total oil. The major dominant constituents were 2-bornanone (28.2%), 1,8-cineole (19.9%), α-thujone (7.5%), β-thujone (6.7%) which accounts for 62.3% of total constituents identified, with volatile oil yield of 4.11% (w/w). The fumigation assay indicated that the volatile oil was acutely toxic to fire ants, with an LC50 of 16.47 µl/l. Among the constituents tested, only (α + β) thujone and 1,8-cineole were toxic, with LC50 of 17.68 and 30.66 µl/ after 12 h of exposure. The volatile oil, (α + β) thujone and 1, 8-cineole showed strong fumigant activity against the red imported fire ant workers in a time- and dose-dependent manner. The volatile oil caused 100% mortality of the red imported fire ant workers, even at the lowest concentration of 20 µl/l after 24 h of exposure. In addition, the volatile oil and 1,8-cineole inhibited acetylcholinesterase activity, while (α + β) thujone inhibited carboxylesterase activity in the fire ant workers. It has been concluded that the volatile oil and some of the compounds from S. brevifolium might be developed as eco-friendly approaches for the control of red imported fire ants.
Resumo em Inglês:ABSTRACT Lichens that are exclusive symbiotic organisms composed of fungus and alga, are considered as a wealthy source of biologically and pharmacologically active small-molecules thanks to the tight metabolic relationship between symbiotic partners. We herein report cytotoxic, anti-angiogenic and apoptotic profile of a lichen derived small-molecule named parietin. Parietin was isolated from the acetone extract of Xanthoria parietina (L.) Th.Fr (1860), Teloschistaceae, which was gathered from Afyon, Turkey. AlamarBlue™ cell viability, lactate dehydrogenase membrane leakage and PicoGreen™ dsDNA quantitation assays were used to determine the cytotoxic concentrations of parietin on cisplatin-resistant BRCA2-mutated human breast TNM stage IV adenocarcinoma (HCC1428), human breast ductal carcinoma (T-47D), and human umbilical vein endothelial (HUVEC) cells. Additionally, cell adhesion, endothelial tube formation, reactive oxygen species accumulation and active caspase 3 determination assays were employed to identify the anti-angiogenic and apoptotic efficiency of parietin. Low concentrations of parietin such as 50 and 100 µM showed a significant anti-angiogenic and apoptotic activity though the half-maximal inhibitory concentration (IC50) values were higher than 600 µM on the cells. On the other hand, it was observed that parietin shows less cytotoxic and membrane degradative activities on healthy HUVEC cells than the HCC1428 and T-47D breast cancer cells. Parietin seems to be a promising anti-angiogenic and apoptotic lichen metabolite for the further investigations.
Resumo em Inglês:ABSTRACT Various extracts obtained from the red alga Plocamium brasiliense (Greville Howe & Taylor), including a fraction containing crude 5-chloro-1-(E)-chlorovinyl-2,4-dibromo-1,5-dimethylcyclohexane (1) and another containing a mixture of halogenated monoterpenes (F), as well as atomaric acid meroditerpene (2) isolated from brown alga Stypopodium zonale (J. V. Lamouroux) Papenfuss, were evaluated for their activity against Trypanosoma cruzi. The cytotoxic and trypanosomicidal effects of these extracts were evaluated in Vero cells and clinically relevant forms of T. cruzi (amastigotes and trypomastigotes). All extracts from P. brasiliense presented low cytotoxicity and moderate trypanosomicidal effects, except for the hydroalcoholic extract. The crude 1 and F fractions had enhanced trypanocidal activity but showed low selectivity. Moreover, atomaric acid (2) was identified as a hit, demonstrating a potent trypanocidal effect reaching an IC50 <10 µM against two different DTU (Yand high selectivity index (<10). These results identify marine natural products as promising candidates against Chagas disease.
Resumo em Inglês:ABSTRACT Persea americana Mill., Lauraceae, commonly known as the avocado, is native to tropical and subtropical regions, including Brazil. From the leaves of P. americana, one previously undescribed flavonol glycoside (1) together with ten known flavonoids (2-11), four megastigmane glycosides (12-15) and two lignans (16-17) were isolated. Their structures were determined by extensive spectroscopic methods including 1D- and 2D-nuclear magnetic resonance and mass spectrometry data. This is the first investigation that reports megastigmane glycoside and lignan classes within the genus Persea. All the isolated compounds have been assessed through the cell survival of larval zebrafish following neomycin-induced damage and the cell viability of a House Ear Institute-Organ of Corti 1 mouse auditory cell line. Among the tested compounds, juglanin (2) and (+)-lyoniresinol (16) showed significant cell regeneration in neomycin-damaged hair cell without cellular toxicity.
Resumo em Inglês:ABSTRACT This study was designed to explore the pharmacokinetic regularity of the plasma concentration, tissue distribution and excretion of orcinol glucoside from aqueous extracts of raw and processed Curculigo orchioides Gaertn., Hypoxidaceae. The experiment first used an ultrahigh-performance liquid chromatography-tandem mass spectrometry approach with multiple reaction monitoring and a positive mode to separate orcinol glucoside from naringin to obtain the plasma concentration curves, bar graph of tissue distribution and excretion curves. These results might be beneficial for reasonable clinical application of C. orchioides and for further development of its wine and salt-processing mechanism.
Resumo em Inglês:ABSTRACT This paper describes the chemical composition and the enantiomer content of the volatile oil hydrodistilled from Clinopodium brownei (Sw.) Kuntze, Lamiaceae. The plant was collected in the South of Ecuador. Thirty one components were identified by GC-MS, which accounted for the 96.15% of the volatile oil. The major components were pulegone (48.44%), menthone (34.55%) and β-acorenol (3.41%). Oxygenated monoterpenes (86.06%), followed by oxygenated sesquiterpenes (5.36%) constituted the most abundant fractions. The enantiomeric compositions of β-pinene, sabinene, 3-octanol, menthone, pulegone and menthyl acetate were determined by enantioselective GC-MS. (-)-Menthone showed the highest enantiomeric excess (ee = 83.4%). In in vitro tests, the volatile oil showed high selective inhibitory activity for butyrylcholinesterase with an IC50, 13.4 ± 1.8 µg/ml. In contrast, it was weakly active against acetylcholinesterase with an IC50 >250 µg/ml.
Resumo em Inglês:ABSTRACT Pentavalent antimonials and amphotericin B remain as the main drugs to treat human leishmaniasis. However, the high toxicity and variable efficacy of treatment have stimulated the search for novel drug candidates. Naturally occurring alkaloids have a long history of antileishmanial activity. Here, we investigate the effects of the β-carboline-1-propionic acid alkaloid isolated from Quassia amara L., Simaroubaceae, against Leishmania amazonensis and Leishmania infatum. The alkaloid was isolated after liquid-liquid fractionation followed by chromatographic purification of the Q. amara methanol extract. The antileishmanial activity was evaluated by the microdilution method, using resazurin as the viability indicator. In addition, annexin and propidium iodide were used to detect parasites undergoing apoptosis. The anti-amastigote activity of the β-carboline-1-propionic acid alkaloid was determined by the infection of RAW 264.7 macrophages. The alkaloid displayed leishmanicidal activity against Leishmania amazonensis and L. infantum promastigotes and intracellular amastigotes with 50% inhibitory concentration ranging from 2.7 ± 0.82 to 9.4 ± 0.5 µg/ml and selectivity indexes >10. Moreover, apoptotic Leishmania amazonensis (19.5%) and L. infantum (40.4%) promastigotes were detected after 5 h incubation with the alkaloid. Finally, the β-carboline-1-propionic acid alkaloid inhibited the production of NO of infected macrophages, suggesting that the intracellular amastigote elimination occurs in a nitrosative stress-independent way. The results shown here suggest that the β-carboline-1-propionic acid alkaloid has potential as an antileishmanial agent.
Resumo em Inglês:ABSTRACT At present, there is a rapidly growing interest in studying the cytotoxic effects of Artemisia herba alba Asso, Asteraceae, in various cancer cell lines. However, its antitumor effectiveness has not been investigated. Therefore, the current study was conducted to study the effect of A. herba alba extract on the proliferation and growth of solid tumor cells in Ehrlich Solid Carcinoma bearing mice. Oral administration of A. herba alba extract resulted in significant reductions in tumor size, tumor weight and mice body weight, as well as caused concurrent significant increases in the DNA breakages and apoptotic DNA damage induction in a time-dependent manner. A. herba alba extract also raised the expression level of p53 gene and reduced of K-ras expression in a time-dependent manner. Minor histological lesions were observed in the liver and kidney tissues sections of mice administered A. herba alba extract compared with the high histological lesions observed in the liver and kidney tissues of artesunate and cisplatin treated groups. Thus, we concluded that A. herba alba extract exhibited promising potential antitumor efficacy with greater safety than artesunate and the commercially used anticancer drug cisplatin in mice.
Resumo em Inglês:ABSTRACT Erythraline is the major alkaloid produced by Erythrina verna Vell., Fabaceae, a plant used in folk medicine and phytotherapeutic products to treat anxiety and sleep disorders. This study aimed to investigate the pharmacokinetic parameters of erythraline after intravenous administration in rats. For the analysis of erythraline in rat plasma, a method was developed and validated using UHPLC-MS/MS. Pharmacokinetic parameters were estimated by non-compartmental analysis. The metabolite 8-oxo-erythraline observed in previous biomimetic model studies was monitored during in vivo experiments. The quantification limit was 5 ng/ml within a linear range of 5-2000 ng/ml. After an intravenous dose of 1 mg/kg, the following pharmacokinetic parameters were observed: elimination half-life of 44.2 min; total clearance of 42.1 ml/min/kg and volume of distribution of 2085 ml/kg. In summary, a precise, accurate and selective UHPLC-MS/MS method was developed and successfully applied to investigate the pharmacokinetics of erythraline in rats. The metabolite 8-oxo-erythraline was observed in rat plasma after 20 min of erythraline administration.
Resumo em Inglês:ABSTRACT Lippia alba (Mill.) N.E.Br. ex Britton & P. Wilson, Verbenaceae, is considered a great source of a bioactive volatile oil. Due to the wide range of known chemotypes, its chemical analysis is very important. Among the several activities of this volatile oil, a potential larvicidal action against Culicidae species is highlighted. However, the low water miscibility of volatile oils limits their application in aqueous media. Oil in water nano-emulsions are in the spotlight of novelty to solve this main problem. Thus, the aim of the present study was to obtain this nanostructured system with L. alba volatile oil (citral chemotype) and evaluate its larvicidal activity against Aedes aegypti and Culex quinquefasciatus larvae. The major compounds were geranial (30.02%) and neral (25.26%). Low mean droplet size (117.0 ± 1.0 nm) and low polydispersity index (0.231 ± 0.004) were observed and no major changes were observed after seven days of storage. LC50 values against C. quinquefasciatus and A. aegypti third-instar larvae were respectively 38.22 and 31.02 ppm, while LC90 values were, respectively, 59.42 and 47.19 ppm. The present study makes use of a low energy, solvent-free and ecofriendly method with reduced costs. Thus, this paper contributes significantly to phyto-nanobiotechnology of larvicidal agents, opening perspectives for the utilization of L. alba volatile oil in integrated practices of vector control.
Resumo em Inglês:ABSTRACT Himalayas are one among the world biodiversity hotspots harboring many endemic medicinal plants. Despite augmentation in the documentation of ethnopharmacological knowledge of medicinal plant species, information regarding endemic species is still underway. Current paper highlights the traditional medicinal uses of rare endemic and unexplored group of plants having potential for novel chemical constituents with effective pharmacological activities. In total, 142 informants (91 male and 51 female) including seventeen traditional healers were interviewed using semi-structured questionnaire, personal observations and group discussions. Interviews were taken in field or otherwise photographs were shown for identification. Females were interviewed indirectly through male family members. For data analysis, quantitative analytical approach was adopted using ethnopharmacological indices as Relative frequency of citations and Fidelity Level. In total, 38 endemic plant species belonging to nineteen families were utilized by the local inhabitants. Highest number of endemics was belonging to Ranunculaceae (7), followed by Gentianaceae and Rosaceae (4 each) with respect to number of species. Highest number of endemics was used in fever, wound healing, throat infection and tonic (4 species each). Root was the most widely used part (36.17%) in cure of diseases and the leading mode administered was decoction (25.49%). Highest use reports and RFC values were recorded for Pimpinella stewartii (58 citations, 0.41 RFC), Caltha alba var. alba (52 citations, 0.37 RFC). Endemic plant species considerably contribute toward ethnomedicinal knowledge and despite rarity, the communities prefer their utilization. Conservation of endemics is necessary for future availability to the local communities.
Resumo em Inglês:ABSTRACT This study compared the influence of mechanical damage and drying on the chemical composition of Mikania glomerata Spreng. and Mikania laevigata Sch.Bip. ex Baker, Asteraceae, leaves. Leaves were collected 1-24 h after damage. Oven-drying at 40ºC and shade-drying at ambient temperature were compared to lyophilization. Samples were extracted in 70% ethanol and analyzed by ultra-high-performance liquid chromatography with mass spectrometry. Significant (p < 0.05) increases of caffeoylquinic acids were observed in damaged leaves of both species and coumarin decreased in M. laevigata, indicating stress. Although the final water content was similar, the drying method affected the leaf composition. In shade-dried leaves of M. laevigata coumarin decreased and the presence of umbelliferone was observed; caffeoylquinic acid contents increased for 288 h in in both species. Apparently, enzymes were inactivated after 6 h in oven drying, stabilizing their chemical composition, while shade drying allowed enzymatic and microbial activity to continue; illustrating the importance of post harvesting procedures on the quality of medicinal plants.
Resumo em Inglês:ABSTRACT Spathulenol was isolated from an extract of Azorella compacta Phil., Apiaceae, by various chromatographic method; identification of the chemical structure was confirmed by comparing its spectroscopic data with those reported in the literature. The anti-Mycobacterium tuberculosis activity of spathulenol was evaluated on MDR, pre-XDR, and XDR clinical isolates of M. tuberculosis, as well as on the reference susceptible strain H37Rv and its cytotoxic activity was evaluated on the Vero Cell Line. The anti-M. tuberculosis activity of spathulenol was twice as potent against the MDR, pre-XDR, and XDR clinical isolates (6.25 µg/ml) than on the susceptible H37Rv strain (12.5 µg/ml). Additionally, the anti-M. tuberculosis activity shown by spathulenol was established as bactericidal on drug-resistant and susceptible strains of M. tuberculosis. Finally, cytotoxic activity on the Vero cell line (CC50 = 95.7 µg/ml) indicated that spathulenol is a selective anti-M. tuberculosis compound, with a selective index of 15.31 against drug-resistant clinical isolates of M. tuberculosis.
Resumo em Inglês:ABSTRACT Plants represent a huge source of substances, with pharmacological potential. Brazil has a diversity of agricultural insect pests and an urgent need for safer methods of insect control. Dysdercus peruvianus (Guerin-Meneville, 1831), Pyrrhocoridae, is an economically important species of the Order Hemiptera and a pest of cotton (Gossypium hirsutum L., Malvaceae). Secondary metabolites in stems of Clusia hilariana Schltdl., Clusiaceae, such as terpenes and benzophenones, have been reported to be insecticidal. The present study investigated the effects of semi-purified fractions of hexane crude extracts from male C. hilariana stems on development of D. peruvianus. Biological parameters at different stages of development including body malformations, range of molting period and toxicity were evaluated. Most insects died and failed to develop due to attachment of their exuviae to the abdomen. Deformations of wings and defective tarsi also occurred. The secondary metabolites from semi-purified fractions of C. hilariana caused mortality, interference in molting and metamorphosis, and body deformations, probably by interacting with the neuroendocrine system. The results demonstrate the potential of C. hilariana extracts as an alternative for the control of the phytophagous insect D. peruvianus and for the development of environmentally safe and biodegradable bio insecticides.
Resumo em Inglês:ABSTRACT Propolis, is a bee product collected from exudates and flower buds of several plants, has strong aroma and several biological applications. This study aimed at evaluating the chemical composition and in vitro antioxidant, antibacterial and cytotoxic properties of volatile oil from Brazilian brown propolis. It was extracted by hydrodistillation and analyzed by gas chromatography-flame ionization detection and gas chromatography-mass spectrometry. Volatile oil from brown propolis exhibited strong antibacterial activity against H. pylori (MIC 3.25 µg/ml), Mycobacterium tuberculosis (MIC 50 µg/ml) and M. avium (MIC 62.5 µg/ml). It was evaluated in vitro for antioxidant activity by DPPH (IC50 25.0 µg/ml) and ABTS (IC50 30.1 µg/ml) methods. Its cytotoxic property was evaluated in normal (human fibroblasts, GM07429A) and tumor (MCF-7-human breast adenocarcinoma; HeLa-human cervical adenocarcinoma and M059J-human glioblastoma) cell lines. IC50 values were 81.32 µg/ml for GM07429A and 85.00, 129.40 and 84.12 µg/ml for MCF-7, HeLa and M059J cells, respectively. Three major dereplicated components of volatile oil from brown propolis were acetophenone (15.2%), nerolidol (13.3%), and spathulenol (11.6%). Our results contribute to a better understanding of the chemical and biological properties of Brazilian brown propolis and provide evidence for its potential medicinal use.
Resumo em Inglês:ABSTRACT The objective of this study was to evaluate the acaricidal activity of the volatile oils of three species of Croton, Euphorbiaceae, against the cattle tick Rhipicephalus microplus. The volatile oils were obtained by hydrodistillation, analyzed by GC-MS and GC-FID and their acaricidal activity was evaluated by the larval packet test and adult immersion test. The volatile oils from Croton conduplicatus Kunth, Croton pulegiodorus Baill., and two different collections of Croton grewioides Baill. (CG1 and CG2) showed eucalyptol (24.09%), p-cymene (23.13%) and methyl chavicol (83.59% and 95.38%) as the major compounds, respectively. All the volatile oils tested in this study showed efficacy against larvae and engorged females of Rhipicephalus microplus. Therefore, Croton pulegiodorus volatile oil is promising for a potential acaricidal formulation because of the best activity against both stages of the cattle tick.