RESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Pfaffia glomerata roots are widely used in Brazil to treat various pathological conditions, particularly psychological disorders. 20-hydroxyecdysone, a phytosteroid present in the plant, can promote greater body resistance against exogenous and endogenous stressors. The objective of this study was to evaluate the possible neuroprotective effect of a 20-hydroxyecdysone-enriched fraction (20E-EF), obtained from P. glomerata roots, in an acute murine stress model. MATERIAL AND METHODS: The 20E-EF was obtained by partitioning the methanol extract from P. glomerata roots with dichloromethane. Mice were treated by gavage with three doses of 20E-EF (3, 10, and 30 mg/kg) and parameters of stress, anxiety, and depression were evaluated. Biomarkers of oxidative stress (enzymes, antioxidant profile, and oxidized molecules) were evaluated in the cortex, striatum (basal ganglia), and hippocampus of animals treated with 30 mg/kg of 20E-EF. RESULTS: Mass spectrometry revealed that 20E was the main compound in the dichloromethane fraction. At a dose of 30 mg/kg, 20E-EF reduced stress, anxiety, and depression, while stimulating antioxidant enzymes (catalase, superoxide dismutase, and glutathione peroxidase), promoting antioxidant activity (antioxidant capacity, sulfhydryl groups, and reduced glutathione), and reducing oxidative markers (lipid peroxidation). In addition, 20E increased the concentration of NO in the striatum, possibly improving memory function and antioxidant activity. CONCLUSION: A 30 mg/kg dose of 20E-EF was able to reduce stress, anxiety, and depression, in addition to maintaining antioxidant defenses of the cortex and striatum. These findings open new perspectives for understanding the therapeutic properties of P. glomerata and the underlying mechanism(s).
Assuntos
Amaranthaceae , Ansiolíticos/farmacologia , Antidepressivos/farmacologia , Ansiedade/prevenção & controle , Comportamento Animal/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Depressão/prevenção & controle , Ecdisterona/farmacologia , Extratos Vegetais/farmacologia , Raízes de Plantas , Estresse Psicológico/prevenção & controle , Amaranthaceae/química , Animais , Ansiolíticos/isolamento & purificação , Antidepressivos/isolamento & purificação , Antioxidantes/farmacologia , Ansiedade/metabolismo , Ansiedade/fisiopatologia , Ansiedade/psicologia , Biomarcadores/metabolismo , Encéfalo/metabolismo , Encéfalo/fisiopatologia , Depressão/metabolismo , Depressão/fisiopatologia , Depressão/psicologia , Modelos Animais de Doenças , Ecdisterona/isolamento & purificação , Comportamento Exploratório/efeitos dos fármacos , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Memória/efeitos dos fármacos , Camundongos Endogâmicos C57BL , Atividade Motora/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/isolamento & purificação , Raízes de Plantas/química , Estresse Psicológico/metabolismo , Estresse Psicológico/fisiopatologia , Estresse Psicológico/psicologiaRESUMO
: Deep eutectic solvents (DESs) were used in combination with macroporous resins to isolate and purify flavonoids and 20-hydroxyecdysone from Chenopodium quinoa Willd by preparative high-performance liquid chromatography (HPLC). The extraction performances of six DESs and the adsorption/desorption performances of five resins (AB-8, D101, HPD 400, HPD 600, and NKA-9) were investigated using the total flavonoid and 20-hydroxyecdysone extraction yields as the evaluation criteria, and the best-performing DES (choline chloride/urea, DES-6) and macroporous resin (D101) were further employed for phytochemical extraction and DES removal, respectively. The purified extract was subjected to preparative HPLC, and the five collected fractions were purified in a successive round of preparative HPLC to isolate three flavonoids and 20-hydroxyecdysone, which were identified by spectroscopic techniques. The use of a DES in this study significantly facilitated the preparative-scale isolation and purification of polar phytochemicals from complex plant systems.
Assuntos
Chenopodium quinoa/química , Cromatografia Líquida de Alta Pressão/métodos , Ecdisterona/isolamento & purificação , Flavonoides/isolamento & purificação , Extratos Vegetais/isolamento & purificação , Adsorção , Cromatografia Líquida de Alta Pressão/instrumentação , Resinas Sintéticas/químicaRESUMO
The crude methanol extract of Sphenocentrum jollyanum root exhibited 98% and 80% antimicrobial activity against Aspergillus fumigatus Pinh and Vancomycin resistant enterococcus (VRE) at a concentration of 200⯵g/mL, with IC50 11.45 and 12.95⯵g/mL, respectively. The ethyl acetate fraction of methanol extract showed in-vitro antimicrobial activity against A. fumigatus Pinh at 83% with IC50 ofâ¯<8⯵g/mL. The phytochemical investigation of ethyl acetate fraction yielded six compounds, which were identified by their NMR, IR and MS spectral analyses as two new phytoecdysteroidal glycosides Sphenocentroside A (1), and Sphenocentroside B (2), and four known phytoecdysteroids: polypodoaurein (3), polypodine B (4), ecdysterone (5), and 20, 26-dihydroxyecdysone (6).
Assuntos
Antibacterianos/farmacologia , Antifúngicos/farmacologia , Ecdisterona/farmacologia , Menispermaceae/química , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Raízes de Plantas/química , Antibacterianos/química , Antibacterianos/isolamento & purificação , Antifúngicos/química , Antifúngicos/isolamento & purificação , Aspergillus fumigatus/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ecdisterona/química , Ecdisterona/isolamento & purificação , Testes de Sensibilidade Microbiana , Conformação Molecular , Extratos Vegetais/isolamento & purificação , Estereoisomerismo , Relação Estrutura-Atividade , Enterococos Resistentes à Vancomicina/efeitos dos fármacosRESUMO
In this work, the aqueous extract obtained from Brazilian ginseng (Pfaffia glomerata) roots (BGR), rich in beta-ecdysone and fructooligosaccharides (FOS), was powdered by spray drying and freeze drying techniques aiming to obtain a novel functional food product. The effects of these drying techniques on the chemical and nutritional quality, morphological and redispersion properties of the BGR powders were evaluated. The BGR powders obtained by both spray drying and freeze drying techniques maintained their beta-ecdysone and FOS contents after drying, demonstrating the stability of these functional compounds. It was found that the wettability of the powders obtained by different treatments was affected by the drying technique because freeze-dried particles reached the lower values (66⯱â¯5â¯s) while spray-dried particles showed a greater time for dispersion into water (150⯱â¯25â¯s). This behavior was mainly associated with differences between powder morphological properties since the freeze-dried particles presented a more porous structure, resulting in a greater water diffusivity into microstructure during the redispersion process. Drying process did not affect the storage stability of powders because the glass transition temperature (Tg) for both samples was approximately 160⯰C at a relative humidity of 56%. Thus, both BGR powders presented adequate redispersion properties to constitute a new functional tea or even to be used as a functional ingredient in food products.
Assuntos
Bebidas , Ecdisterona/isolamento & purificação , Manipulação de Alimentos/métodos , Alimento Funcional , Valor Nutritivo , Oligossacarídeos/isolamento & purificação , Panax/química , Extratos Vegetais/isolamento & purificação , Raízes de Plantas/química , Aerossóis , Cristalografia por Raios X , Liofilização , Tamanho da Partícula , Pós , Solubilidade , Temperatura de Transição , Vitrificação , Água/análise , MolhabilidadeRESUMO
Sida tuberculata (ST) is a Malvaceae species widely distributed in Southern Brazil. In traditional medicine, ST has been employed as hypoglycemic, hypocholesterolemic, anti-inflammatory and antimicrobial. Additionally, this species is chemically characterized by flavonoids, alkaloids and phytoecdysteroids mainly. The present work aimed to optimize the extractive technique and to validate an UHPLC method for the determination of 20-hydroxyecdsone (20HE) in the ST leaves. Box-Behnken Design (BBD) was used in method optimization. The extractive methods tested were: static and dynamic maceration, ultrasound, ultra-turrax and reflux. In the Box-Behnken three parameters were evaluated in three levels (-1, 0, +1), particle size, time and plant:solvent ratio. In validation method, the parameters of selectivity, specificity, linearity, limits of detection and quantification (LOD, LOQ), precision, accuracy and robustness were evaluated. The results indicate static maceration as better technique to obtain 20HE peak area in ST extract. The optimal extraction from surface response methodology was achieved with the parameters granulometry of 710â¯nm, 9â¯days of maceration and plant:solvent ratio 1:54 (w/v). The UHPLC-PDA analytical developed method showed full viability of performance, proving to be selective, linear, precise, accurate and robust for 20HE detection in ST leaves. The average content of 20HE was 0.56% per dry extract. Thus, the optimization of extractive method in ST leaves increased the concentration of 20HE in crude extract, and a reliable method was successfully developed according to validation requirements and in agreement with current legislation.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Ecdisterona/análise , Ecdisterona/isolamento & purificação , Malvaceae/química , Folhas de Planta/química , Ecdisterona/química , Limite de Detecção , Modelos Lineares , Reprodutibilidade dos TestesRESUMO
Glutamate-induced excitotoxicity is a key pathological mechanism in many neurological disease states. Ecdysterones derived from Rhaponticum carthamoides (Willd.) Iljin (RCI) have been shown to alleviate glutamate-induced neuronal damage; although their mechanism of action is unclear, some data suggest that they enhance signaling in the mechanistic target of rapamycin (mTOR) signaling pathway. This study sought to elucidate the mechanisms underlying ecdysterone-mediated neuroprotection. We used in silico target prediction and simulation methods to identify putative ecdysterone binding targets, and to specifically identify those that represent nodes where several neurodegenerative diseases converge. We then used histological analyses in a rat hippocampal excitotoxicity model to test the effectiveness of ecdysterones in vivo. We found that RCI-derived ecdysterones should bind to glutamatergic NMDA-type receptors (NMDARs); specifically, in vivo modeling showed binding to the GRIN2B subunit of NMDARs, which was found also to be a node of convergence in several neurodegenerative disease pathways. Computerized network construction by using pathway information from the Kyoto Encyclopedia of Genes and Genomes (KEGG) database showed putative links between GRIN2B and mTOR pathway elements including phosphoinositide-3kinase (PI3K), mTOR, and protein kinase C (PKC); these elements are associated with neuronal survival. Brain tissue western blots of ecdysterone-treated rats showed upregulated PI3K, Akt, mTOR, and phosphorylated Akt and mTOR, and down regulated GRIN2B and the apoptotic enzyme cleaved caspase-3. Ecdysterone treatment also prevented glutamate-induced rat hippocampal cell loss. In summary, RCI-derived ecdysterones appear to prevent glutamatergic excitotoxicity by increasing mTOR/Akt/PI3K signaling activity.
Assuntos
Ecdisterona/farmacologia , Hipocampo/efeitos dos fármacos , Leuzea/química , Fármacos Neuroprotetores/farmacologia , Transdução de Sinais/efeitos dos fármacos , Serina-Treonina Quinases TOR/metabolismo , Animais , Caspase 3/metabolismo , Ecdisterona/isolamento & purificação , Ácido Glutâmico/farmacologia , Hipocampo/citologia , Masculino , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Estrutura Molecular , Fármacos Neuroprotetores/isolamento & purificação , Fosforilação , Raízes de Plantas/química , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores de N-Metil-D-Aspartato/metabolismo , Regulação para CimaRESUMO
Phytoecdysteroids like 20-hydroxyecdysone ("ecdysterone") can exert a mild, non-hormonal anabolic/adaptogenic activity in mammals, and as such, are frequently used in food supplements. Spinach is well-known for its relatively low ecdysteroid content. Cyanotis arachnoidea, a plant native in China, is among the richest sources of phytoecdysteroids, and extracts of this plant are marketed in tons per year amounts via the internet at highly competitive prices. Here we report the investigation of a series of food supplements produced in Germany and claimed to contain spinach extracts. Twelve ecdysteroids including two new compounds were isolated and utilized as marker compounds. A comparative analysis of the products with Cyanotis and spinach extracts provides evidence that they were manufactured from Cyanotis extracts instead of spinach as stated. Based on the chromatographic fingerprints, 20-hydroxyecdysone 2- and 3-acetate are suggested as diagnostic markers for related quality control. This case appears to represent an unusual type of dietary supplement counterfeiting: undeclared extracts from alternative plants would supposedly 'guarantee' product efficacy.
Assuntos
Commelinaceae/química , Suplementos Nutricionais/normas , Ecdisteroides/análise , Spinacia oleracea/química , Animais , China , Ecdisona/análise , Ecdisona/isolamento & purificação , Ecdisteroides/isolamento & purificação , Ecdisterona/análise , Ecdisterona/isolamento & purificação , Alemanha , Fitosteróis/análise , Fitosteróis/isolamento & purificação , Extratos Vegetais/química , Controle de QualidadeRESUMO
Increasing the activation of protein kinase B (Akt) has been suggested as a key signaling step in the nonhormonal anabolic activity of the phytoecdysteroid 20-hydroxyecdysone (20E) in mammals. Base-catalyzed autoxidation of this compound was shown previously to yield interesting B-ring-modified analogues. Herein is reported a thorough study on this reaction, resulting in the preparation and complete NMR spectroscopic assignments of calonysterone (5) and its previously overlooked desmotropic pair (7), along with two new sensitive metabolites of 20E. The two isomers showed considerable stability in solution. Time dependency of the reaction for yield optimization is also presented; by means of analytical HPLC, the two desmotropes can reach a maximum combined yield of >90%. The activity of these compounds on Akt phosphorylation was tested in murine skeletal muscle cells. Compounds 2 and 5 showed more potent activity than 20E in increasing Akt activation, while compound 7 exerted an opposite effect. As such, the present study provides the first direct evidence for a pair of desmotropes exerting significantly different bioactivities.
Assuntos
Commelinaceae/química , Medicamentos de Ervas Chinesas/química , Ecdisterona/metabolismo , Músculo Esquelético/metabolismo , Animais , Cromatografia Líquida de Alta Pressão , Medicamentos de Ervas Chinesas/isolamento & purificação , Medicamentos de Ervas Chinesas/farmacologia , Ecdisterona/isolamento & purificação , Camundongos , Estrutura Molecular , Fibras Musculares Esqueléticas/metabolismo , Músculo Esquelético/efeitos dos fármacos , Oxirredução , Fosforilação , Raízes de Plantas/química , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de SinaisRESUMO
INTRODUCTION: Ajuga turkestanica is a plant used in traditional medicine for its high ecdysteroid content, including the presence of the particularly active turkesterone, which possess efficient anabolic activity. OBJECTIVES: To isolate and identify minor ecdysteroids present in a semi-purified plant fraction containing ca. 70% turkesterone. MATERIAL AND METHODS: Multi-step preparative HPLC (combining RP- and NP-HPLC systems) was used to purify the different components present in the turkesterone fraction. Isolated compounds were identified by high-resolution mass spectrometry and 2D-NMR. RESULTS: Fourteen ecdysteroids (including turkesterone and 20-hydroxyecdysone) were isolated. Seven of these, all bearing an 11α-hydroxy group, were previously unreported. CONCLUSION: Ajuga turkestanica ecdysteroids are characterised by the abundance of 11α-hydroxylated compounds and by the simultaneous presence of 24C, 27C, 28C and 29C ecdysteroids. It is expected that even more ecdysteroids are to be found in this plant since the starting material for this study lacked the less polar ecdysteroids. The simultaneous presence of 20-hydroxyecdysone and turkesterone (its 11α-hydroxy analogue) as the two major ecdysteroids suggests that every ecdysteroid is probably present in both 11α-hydroxy and 11-deoxy forms.
Assuntos
Ajuga/química , Ecdisteroides/análise , Raízes de Plantas/química , Plantas Medicinais/química , Cromatografia Líquida de Alta Pressão/métodos , Ecdisteroides/química , Ecdisteroides/isolamento & purificação , Ecdisterona/análogos & derivados , Ecdisterona/análise , Ecdisterona/química , Ecdisterona/isolamento & purificação , Espectroscopia de Ressonância Magnética/métodos , Espectrometria de Massas/métodosRESUMO
We investigated the resistance of erythrocytes from rat brain venous blood to acid hemolysis in the dynamics of brain ischemic period (15, 30, 45 and 60 min), as well as in the early (5 min) and distant (24h) period of brain reperfusion. Brain ischemia-reperfusion was made in rats that received ecdysterone (standartized extract of Serratula coronata) within 18 days (per os, 1 mg/kg). Analysis of the kinetic curves of acid hemolysis showed a pronounced (60 times, from 1.45 to 85.85% at 60 min of brain ischemia and at 5 min of brain reperfusion, respectively) increase of unstable erythrocytes that hemolyzed easily (< 2.5 min). In the preconditioned rats, this increase was only 8-fold. During the period of brain ischemia, with a maximum at 15th minute, in the venous blood from brain the diene conjugates (DK) pools increased from 2.40 to 9.48 ng/mg protein and LTC4 pools increased from 1.49 to 5.98 pmol/mg protein. Even more pools of DC and LTC4 were increased at 5th min of brain reperfusion. In animals received ecdysterone, during ischemia and early reperfusion period, both pools of DC and LTC4 in venous blood were lower than that in the controls. The latter implies a possible antiradical mechanism of the protective effect of ecdysterone.
Assuntos
Antioxidantes/farmacologia , Asteraceae/química , Isquemia Encefálica/prevenção & controle , Ecdisterona/farmacologia , Fármacos Neuroprotetores/farmacologia , Traumatismo por Reperfusão/prevenção & controle , Alcenos/sangue , Animais , Antioxidantes/isolamento & purificação , Encéfalo/irrigação sanguínea , Encéfalo/metabolismo , Encéfalo/patologia , Isquemia Encefálica/sangue , Isquemia Encefálica/patologia , Células Cultivadas , Ecdisterona/isolamento & purificação , Eritrócitos/efeitos dos fármacos , Hemólise/efeitos dos fármacos , Precondicionamento Isquêmico , Leucotrieno C4/sangue , Fármacos Neuroprotetores/isolamento & purificação , Fragilidade Osmótica/efeitos dos fármacos , Extratos Vegetais/química , Ratos , Ratos Wistar , Traumatismo por Reperfusão/sangue , Traumatismo por Reperfusão/patologiaRESUMO
Tetra-acetylajugasterone C (TAAC) was found to be one of the naturally occurring compounds of the Cameroonian medicinal plant Vitex cienkowskii which is responsible for a vasorelaxant activity of an extract of this plant. The evaluation of the underlying mechanisms for the relaxing effect of TAAC was determined using aortic rings of rats and mice. TAAC produced a concentration-dependent relaxation in rat artery rings pre-contracted with 1µM noradrenaline (IC50: 8.40µM) or 60mM KCl (IC50: 36.30µM). The nitric oxide synthase inhibitor l-NAME (100µM) and the soluble guanylate cyclase inhibitor ODQ (10µM) significantly attenuated the vasodilatory effect of TAAC. TAAC also exerted a relaxing effect in aorta of wild-type mice (cGKI(+/+); IC50=13.04µM) but a weaker effect in aorta of mice lacking cGMP-dependent protein kinase I (cGKI(-/-); IC50=36.12µM). The involvement of calcium channels was studied in rings pre-incubated in calcium-free buffer and primed with 1µM noradrenaline prior to addition of calcium to elicit contraction. TAAC (100µM) completely inhibited the resulting calcium-induced vasoconstriction. The same concentration of TAAC showed a stronger effect on the tonic than on the phasic component of noradrenaline-induced contraction. This study shows that TAAC, a newly detected constituent of Vitex cienkowskii contributes to the relaxing effect of an extract of the plant. The effect is partially mediated by the involvement of the NO/cGMP pathway of the smooth muscle but additionally inhibition of calcium influx into the cell may play a role.
Assuntos
Ecdisterona/análogos & derivados , Endotélio Vascular/efeitos dos fármacos , Extratos Vegetais/farmacologia , Vasoconstrição/efeitos dos fármacos , Vasodilatação/efeitos dos fármacos , Vasodilatadores/farmacologia , Vitex/química , Animais , Aorta/efeitos dos fármacos , Cálcio/metabolismo , Canais de Cálcio/metabolismo , GMP Cíclico/metabolismo , Relação Dose-Resposta a Droga , Ecdisterona/isolamento & purificação , Ecdisterona/farmacologia , Endotélio Vascular/metabolismo , Inibidores Enzimáticos/farmacologia , Guanilato Ciclase/antagonistas & inibidores , Camundongos , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase/antagonistas & inibidores , Norepinefrina/farmacologia , Casca de Planta , Extratos Vegetais/química , Caules de Planta , Ratos , Receptores Citoplasmáticos e Nucleares/antagonistas & inibidores , Guanilil Ciclase Solúvel , Vasodilatadores/isolamento & purificaçãoRESUMO
Phytoecdysteroid glucosides, brainesterosides A-E, were isolated from the rhizomes of Brainea insignis along with three known phytoecdysteroids, ponasteroside A, ponasterone A, and 20-hydroxyecdysone. Their structures were elucidated by spectroscopic and chemical means. A possible biogenetic pathway is postulated for these compounds. The chemosystematic significance of ponasterone A is discussed.
Assuntos
Medicamentos de Ervas Chinesas/isolamento & purificação , Ecdisterona/análogos & derivados , Glucosídeos/isolamento & purificação , Medicamentos de Ervas Chinesas/química , Ecdisterona/química , Ecdisterona/isolamento & purificação , Glucosídeos/química , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Rizoma/químicaRESUMO
OBJECTIVE: To study the chemical constituents from the roots of Tinospora sagittata var. yunnanensis. METHODS: Various chromatographic techniques were used to isolate and purify the constituents. The structures of these compounds were elucidated on the basis of spectral analysis. RESULTS: Seven compounds were isolated from the plant and their structures were identified as tinophylloloside (1), epitinophylloloside (2), 2-deoxycrustecdysone (3), polypodine B (4) , (+)-5'-methoxyisolariciresinol 3alpha-O-beta-D-glucopyranoside (5), geniposide (6), adenine (7). CONCLUSION: All compounds are isolated from the plant for the first time,and compounds 4, 5, 6 and 7 are isolated firstly from the genus.
Assuntos
Adenina/isolamento & purificação , Ecdisterona/análogos & derivados , Iridoides/isolamento & purificação , Raízes de Plantas/química , Tinospora/química , Adenina/química , Ecdisterona/química , Ecdisterona/isolamento & purificação , Iridoides/química , Espectroscopia de Ressonância Magnética , Plantas Medicinais/química , Plantas Medicinais/crescimento & desenvolvimento , Tinospora/crescimento & desenvolvimentoRESUMO
Bio-guided fractionation of the roots of Paris polyphylla (Trilliaceae), based on inhibition of P-glycoprotein-mediated daunorubicin efflux in K562/R7 cell line, led to isolation and identification of the three saponins 3-O-Rha(1-->2)[Ara(1-->4)]Glc-pennogenine, gracillin and polyphyllin D, and the two ecdysteroids 20-hydroxyecdysone and pinnatasterone. These compounds were tested for multidrug reversion on P-glycoprotein (ABCB1) with both drug-selected and transfected cell lines, and also on Breast Cancer Resistance Protein (BCRP/ABCG2). By contrast to a weak efficiency on BCRP, the three saponins displayed significant effects as inhibitors of P-glycoprotein-mediated drug efflux.
Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/antagonistas & inibidores , Transportadores de Cassetes de Ligação de ATP/antagonistas & inibidores , Daunorrubicina/metabolismo , Proteínas Associadas à Resistência a Múltiplos Medicamentos/antagonistas & inibidores , Proteínas de Neoplasias/antagonistas & inibidores , Extratos Vegetais/farmacologia , Saponinas/farmacologia , Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP , Ciclosporina/farmacologia , Diosgenina/análogos & derivados , Diosgenina/isolamento & purificação , Diosgenina/farmacologia , Resistência a Múltiplos Medicamentos/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Ecdisteroides/isolamento & purificação , Ecdisteroides/farmacologia , Ecdisterona/isolamento & purificação , Ecdisterona/farmacologia , Humanos , Imunossupressores/farmacologia , Células K562 , Leucemia/tratamento farmacológico , Leucemia/metabolismo , Magnoliopsida/química , Moduladores de Transporte de Membrana/farmacologia , Extratos Vegetais/química , Rizoma , Saponinas/isolamento & purificação , Espirostanos/isolamento & purificação , Espirostanos/farmacologia , Esteroides/isolamento & purificação , Esteroides/farmacologiaRESUMO
INTRODUCTION: The genus Asparagus is known to contain phytoecdysteroids that have been shown to exhibit many beneficial pharmacological properties such as improving lipid metabolism, modulating immunological responses, etc. Currently, knowledge about the contents of phytoecdysteroids in the roots of Asparagus species is limited and HPLC methods for their analyses are unsatisfactory. OBJECTIVE: To develop an HPLC method for the simultaneous determination of three phytoecdysteroids, 20-hydroxyecdysone, ecdysone and ajugasterone C, in the roots of four Asparagus species. METHODOLOGY: Reference standards of phytoecdysteroids were isolated from the roots of Asparagus filicinus by open column chromatography. HPLC analysis was performed on an Alltima C(18) column with gradient elution using aqueous 0.2% formic acid solution containing 0.2% isopropanol and acetonitrile. RESULTS: All calibration curves showed good linear correlation coefficients (r(2) > 0.9994) within the tested ranges. Limits of detection (S/N = 3) and quantification (S/N = 10) for the three analytes were less than 2.7 and 9.9 ng, respectively. Intra- and inter-day RSDs of retention times and peak areas were less than 2.61%. The recoveries were between 93.2 and 107.5%, and the RSDs were less than 3.83% for the root samples of A. filicinus. CONCLUSION: The HPLC method established is appropriate for the efficient quantitative and qualitative analyses of important phytoecdysteroids in Asparagus species. This study showed that A. filicinus is rich in phytoecdysteroids, especially 20-hydroxyecdysone. However the three studied phytoecdysteroids were not detected in A. cochinchinensis, A. officinalis and A. setaceus.
Assuntos
Asparagus/química , Ecdisteroides/química , Calibragem , Cromatografia Líquida de Alta Pressão , Ecdisona/química , Ecdisona/isolamento & purificação , Ecdisterona/análogos & derivados , Ecdisterona/química , Ecdisterona/isolamento & purificação , Raízes de Plantas/química , Plantas Medicinais/química , Padrões de Referência , Reprodutibilidade dos Testes , Solventes , Especificidade da Espécie , Espectrofotometria UltravioletaRESUMO
Leuzea carthamoides is an adaptogenic plant containing biologically active compounds as ecdysteroids and guaianolide-type sesquiterpene lactones, conventionally extracted from the plant with ethanol. It may be a potential source of the mentioned natural compounds. Ethanol-modified near-critical CO(2) was used as selective solvent with the aim to increase the level of 20-hydroxyecdysone in the extract from L. carthamoides roots and to remove selectively cynaropicrin, a sesquiterpene lactone of bitter taste, from the leaves. The extraction conditions were varied (pressure 20-28 MPa, temperature 40-60 degrees C, ethanol concentration in the solvent 0-7.1%) and the extraction yield and extract composition were compared with the results of ethanolic extraction. The supercritical fluid extraction (SFE) from finely powdered plant was controlled by phase equilibrium. Cynaropicrin was quantitatively removed from the leaves where 89% of 20-hydroxyecdysone was retained. The extraction yield of 20-hydroxyecdysone from roots with ethanol-modified CO(2 )was lower by 30% than with ethanol but its concentration in the extract was higher by 67%.
Assuntos
Cromatografia com Fluido Supercrítico/métodos , Ecdisterona/isolamento & purificação , Lactonas/isolamento & purificação , Leuzea/metabolismo , Sesquiterpenos/isolamento & purificação , Técnicas de Química Analítica/métodos , Ecdisterona/análise , Etanol/química , Lactonas/análise , Modelos Químicos , Extratos Vegetais/metabolismo , Folhas de Planta/metabolismo , Raízes de Plantas/metabolismo , Pressão , Sesquiterpenos/análise , Solventes/química , TemperaturaRESUMO
Ecdysterone has been found in a great many plants and animals and has some valuable pharmaceutical properties. The present study was conducted to investigate optimal conditions for the extraction of the compound by supercritical fluid extraction from the roots of Achyranthes bidentata BL. An orthogonal array design (OAD), OA(9)(3(4)), was employed as a chemometric method for optimization of the extraction of ecdysterone from the herbal medicine. Four parameters, namely, pressure and temperature of the supercritical fluid, the dynamic extraction time, and the flow rate of dimethyl sulfoxide, were studied and optimized by a three-level OAD. Determinations of the extracts were performed by high-performance liquid chromatography. The effects of the parameters were studied using analysis of variance. The results shown that the yield of ecdysterone could be influenced by the four parameters to a similar degree. The yield for DMSO-modified supercritical CO(2) was in the range from 0.65 to 1.03 mg/g under the selected conditions. In comparison with methanol-modified supercritical CO(2 )and Soxhlet extraction, a higher yield was obtained when DMSO-modified supercritical CO(2) was used.
Assuntos
Achyranthes/metabolismo , Cromatografia com Fluido Supercrítico/métodos , Ecdisterona/isolamento & purificação , Extratos Vegetais/análise , Raízes de Plantas/metabolismo , Análise de Variância , Dióxido de Carbono/química , Técnicas de Química Analítica/métodos , Cromatografia/métodos , Cromatografia Líquida de Alta Pressão/métodos , Ecdisterona/análise , Ecdisterona/química , Modelos Químicos , Plantas Medicinais/metabolismo , Pressão , Temperatura , Fatores de TempoRESUMO
OBJECTIVE: To study the non-taxoids in the leaves of Taxus mairei. METHOD: The chemical constituents were isolated by chromatography and identified by spectral data. RESULT: Five compounds, taxamairin A (1), taxamairin B (2), sciadopitysin (3), ( - ) matairesinol (4), ponasterone A (5) were isolated and identified. CONCLUSION: Compounds 3-5 were isolated from this plant for the first time, compounds 1 and 2 were isolated from the leaves of T. mairei for the first time.
Assuntos
Biflavonoides/isolamento & purificação , Ecdisterona/análogos & derivados , Furanos/isolamento & purificação , Lignanas/isolamento & purificação , Plantas Medicinais/química , Taxus/química , Biflavonoides/química , Diterpenos/química , Diterpenos/isolamento & purificação , Ecdisterona/química , Ecdisterona/isolamento & purificação , Furanos/química , Lignanas/química , Folhas de Planta/químicaRESUMO
OBJECTIVE: To study the chemical constituents from Lamium maculatum var. kansuense. METHOD: The chemical constituents were isolated and repeatedly purified on silica gel column and the structures were elucidated by the NMR spectra and physico-chemical properties. RESULT: Six compounds were obtained and identified as polypodine B (I), 5-OH-8-epiloganin (II), shlanzhiside methyl ester (III), liriodendrin (IV), quercitroside (V), uridine (VI). CONCLUSION: Compound IV was found from genus Lamium for the first time and the rest of the compounds were found from Lamium maculatum var kansuense for the first time.
Assuntos
Ecdisterona/análogos & derivados , Ecdisterona/isolamento & purificação , Furanos/isolamento & purificação , Glucosídeos/isolamento & purificação , Lamiaceae/química , Plantas Medicinais/química , Ecdisterona/química , Furanos/química , Glucosídeos/química , Uridina/química , Uridina/isolamento & purificaçãoRESUMO
OBJECTIVE: To study the water-soluble chemical constituents in root of Achyranthes bidentata. METHOD: The chemical constituents were isolated by silica gel column chromatography and the structures were elucidated by the NMR spectra and physico-chemical properties. RESULT: Seven compounds were obtained and identified as n-butyl-beta-D-fructopyranoside (I), oleanoic acid (II), 3-O-[beta-D-glucopyranosyl], oleanoic acid 28-O-beta-D-glucopyranoside (III), allantoin (IV), 20-hydroxy ecdysone (V), glutamic acid (VI), 3-O-[beta-D-glucopyranosyl], oleanoic acid 28-O-beta-D-glucopyranosyl ester (VII). CONCLUSION: Compounds III-VII were obtained from this plant for the first time.