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Abstract Hebanthe eriantha (Martius) Kuntze and Pfaffia glomerata (Spreng) Pedersen are medicinal plants popularly known as "Brazilian Ginseng" due to their similarity to Panax ginseng. In Brazil, they are sold as the same herb, despite their different pharmacological and toxicological properties. The morphological identification is difficult, which facilitates their adulteration. We report the application of the Barcode DNA High-Resolution Melting (Bar-HRM) using matK gene to differentiate both species in samples sold in the Brazilian market. Using the proposed method, we could discriminate and identify both species. Bar-HRM analysis allowed discriminating and identifying both species. It allowed the identification of H. eriantha and P. glomerata in 43.6% and 56.4% of the amplified samples, respectively. Of these, only seven samples were authenticated and, in 71.4% of the cases, adulterated. We concluded that Bar-HRM has proven to be a fast alternative method to authenticate plants under the common name "Brazilian Ginseng".
Subject(s)
Amaranthaceae/classification , Panax/classification , Plants, Medicinal/adverse effectsABSTRACT
ABSTRACT: Plants that contain antioxidant compounds have attracted increasing interest for their vital role in the attenuation of oxidative damage caused by free radicals and in the treatment of various diseases. The present study investigated the β-ecdysone content and the antioxidant activity of Brazilian ginseng (Pfaffia glomerata) extracts obtained from inflorescences, stems, and roots. The P. glomerata extracts were tested for antioxidant activity using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging method, β-carotene bleaching test, and phosphomolybdenum method. The β-ecdysone content of P. glomerata extracts was measured by high-performance liquid chromatography (HPLC). The P. glomerata inflorescences showed the strongest DPPH radical scavenging activity and the strongest antioxidant activity in the β-carotene bleaching assay and phosphomolybdenum test. The roots showed the lowest antioxidant capacity in all of the assays. The concentration of β-ecdysone in the plant organs followed the following decreasing order: inflorescences > stems > roots. The present study showed that P. glomerata inflorescence extract had high antioxidant capacity that could be attributed to the presence of β-ecdysone.
RESUMO: Plantas que contêm compostos antioxidantes têm atraído interesse crescente por seu papel fundamental na atenuação de danos oxidativos causados pelos radicais livres e no tratamento de várias doenças. O presente estudo investigou o conteúdo de β-ecdysone e a atividade antioxidante de extratos de ginseng brasileiro (Pfaffia glomerata) obtidos a partir das inflorescências, caules e raízes. Os extratos de Pfaffia glomerata foram testados para atividade antioxidante usando o método sequestrante do radical 2,2-difenil-1-picrilhidrazil (DPPH), sistema modelo β-caroteno-linoleato e método de fosfomolibdênio. O conteúdo de β-ecdisona dos extratos de P. glomerata foi medido por cromatografia líquida de alta eficência (CLAE). As inflorescências de P. glomerata mostraram a maior atividade sequestrante de radical DPPH e a maior atividade antioxidante no ensaio β-caroteno-linoleato e no teste de fosfomolibdênio. As raízes mostraram a menor capacidade antioxidante em todos os ensaios. A concentração de β-ecdisona nos órgãos da planta seguiu a seguinte ordem decrescente: inflorescências > caules > raízes. Os resultados indicaram uma correlação positiva entre conteúdo de β-ecdisona e atividade sequestrante de radical DPPH. O presente estudo mostrou que o extrato das inflorescências de P. glomerata teve alta atividade antioxidante que poderia ser atribuída à presença de β-ecdisona.
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Objective: To study the chemical constituents from the roots of Pfaffia paniculata. Methods: The compounds were isolated and purified by means of chromatographic separation techniques and their structures were identified based on spectral features. Results: Fourteen known compounds, including 11 30-noroleanane triterpenoids and three ecdysterones, named pfaffianol A (1), 3-oxo-akebonoic acid (2), 16β-hydroxyl-3-oxo-akebonoic acid (3), pfaffiaglycoside A (4), pfameric acid (5), 2α,3β,20α-trihydroxy-30-norolean-12-en-28-oic acid (6), 2α,3β-dihydroxy-23-oxo-30-norolean-12,20(29)-dien-28-oic acid (7), 2α,3β-dihydroxy-30-norolean-12,20(29)-dien-28-oic acid (8), 2α,3α-dihydroxy-30-norolean-12,20(29)-dien-28-oic acid (9), quinatic acid (10), 3β-hydroxy-30-norhederagenin (11), diaulusterol B (12), 20,22-didehydrotaxisterone (13), and ecdysterone (14) were isolated from the roots of P. paniculata. Conclusion: Compounds 10-12 are obtained from P. paniculata for the first time. Compounds 6-9, and 13 are isolated from the genus of Pfaffia for the first time.
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Objective: To establish a method for the determination of oleanolic acid(OA)in Pfaffia glomerata to compare the contents of OA in different parts of P. glomerata from Guangxi, China, with different growth years and different habitats. Methods: HPLC method was adopted. The determination was performed on Phenomenex Luna C18(250 mm×4.6 mm, 5 μm)column with mobile phase consisting of methanol-water-glacial acetic acid-triethylamine(285:15: 0.2:0.1, V/V) at the flow rate of 1.0 ml/min. The column temperature was set at 30℃. The detection wavelength was set at 210 nm, and the injection volume was 20 μl. The content of OA in P. glomerata was determined by the established method, and the extraction process of OA was optimized by the orthogonal test. Results: The linear range of OA was 0.0206-2.060 mg/ml(r2=1.0000). RSDs for the precision, repeatability and stability tests were all lower than 2%(n=6 or n=7). The average recovery of OA was 100.89%(RSD=1.69%, n=9). The optimum extraction conditions of OA were as follows: 20-fold ethanol(80%, V/V), extracting for three times, refluxing 1.5 hour each time, and processing acid hydrolysis with 3.0% sulfuric acid(g/g)for 1 hour. Under these conditions, the OA had the highest extraction efficiency. The contents of OA in reed head, root, old stem, tender stem, leaf and flower of P. glomerata were 4.87, 4.61, 2.67, 0.99, 0.24 and 1.13 mg/g, respectively. The average contents of OA in the roots of P. glomerata aged 1, 2, 3, 4 and 5 years in Guangxi were 3.08, 4.07, 4.71, 4.62 and 4.46 mg/g, respectively. Conclusion: The established extraction process and detection method is suitable for the extraction and content determination of OA in P. glomerata. Although OA is distributed in all parts of P. glomerata, the contents significantly vary in different parts. The content of OA is highest in reed head and lowest in leaves. The OA content in P. glomerata becomes stable after 3 years of growth in Guangxi.
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Objective: To establish a macroporous resin enrichment process for total triterpene sapogenins from Pfaffia glomerata,and evaluate the free radical scavenging activity of the total triterpene sapogenins. Methods: The transfer rate of triterpene sapogenins was used as the index. Static adsorption-desorption method was used to select the best macroporous resin,dynamic adsorption-desorption method was used to determine the enrichment process parame- ters,and orthogonal test was used to optimize the enrichment process. The 1,1- diphenyl- 2- trinitrophenylhydrazine (DPPH)and hydroxyl free radical scavenging tests were used to evaluate the free radical scavenging capacity of the triter- pene sapogenins. Results: The AB-8 type macroporous resin had the best enrichment effect on the triterpene sapoge-nins. The conditions for the optimum enrichment process were as follows:the concentration of triterpene sapogenins in the sample solution for resin column chromatography was 3.5 mg/ml,the ratio of diameter to height for the bed of resin column was 1:7,the amount of the raw herbs subjected to the resin column was 0.11 g/ml(herbs/resin),the flow rate for subjecting sample solution to resin column was 1 BV/h,the eluent was 95% aqueous ethanol,the eluting flow rate was 2BV/h,and the eluent volume for elution was 7 BV. After optimization,total triterpene sapogenins reached 65% in the samples prepared by the optimized process,and the transfer rate of total triterpene sapogenins reached 80.23% in the pre- paring process. The total triterpene sapogenin sample could scavenge the DPPH and hydroxyl free radicals,and the scav- enging rate for the DPPH and hydroxyl radicals reached 82.42% and 73.43% at the 1.0 mg/ml,respectively. Conclusion: The AB- 8 macroporous resin can be used for the enrichment and purification of triterpene sapogenins from P. glomerata. The optimized enrichment process was stable and feasible,and the obtained triterpene sapogenins showed a good free radical scavenging activity.
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Objective: To study the chemical constituents of ethyl acetate extract from domestic Pfaffia glomerata. Methods: The compounds were isolated and purified by silica gel, reversed-phase, Sephadex LH-20 column chromatography, and their structures were elucidated by spectroscopic analysis as well as chemical methods. Results: Twenty compounds were identified as 1-undecanol (1), oleic acid (2), β-sitosterol (3), 2-hydroxy-4,6-dimethoxyacetophenone (4), oleanolic acid (5), pfaffianol A (6), benzene-1,4-diol (7), vanillic acid (8), iresinone (9), ethylcaffeate (10), oleanoicaeid-28-O-β-D-glucopyranosyl ester (11), allantoin (12), ajugasterone-C (13), β-ecdysterone (14), iresinoside (15), adenine (16), oleanolic acid 3-O-β-D-glucuronopyranoside (17), ficusoside B (18), pfaffiaglycoside B (19), and β-D-Glucopyranosyl-3-(O-β-D-glucopyranosyloxy)-oleanolate (20). Conclusion: Compounds 7-10, 15, 17, 18, and 20 are isolated from this plant for the first time.
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Objective: To establish and optimize the technique of tissue culture and rapid propagation of Pfaffia paniculata. Methods: Using MS and 1/2MS basic media, the effects of various combinations of plant growth regulators (6-BA, NAA, IBA, and IAA) on the seedlings subculture and rooting culture were studied by orthogonal design. Results: The effective medium for cluster buds-inducing and subculture was MS + 6-BA 1.5 mg/L + NAA 0.1 mg/L + IBA 0.2 mg/L, and the propagation coefficient was over 6.0 per 30 d; The best rooting medium was 1/2 MS + NAA 1.0 mg/L + IBA 0.2 mg/L, and the rooting rate was 100%. The rooting seedlings were transplanted in well-drained sand bed and the survival rate was 98%. Conclusion: The method in the experiment could provide a basis on protecting the mild resources of P. paniculata exploring the artificial resources, and discussing the new way breedings.
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Pfaffia glomerata (Spreng.) Pedersen, Amaranthaceae, is widely distributed in Brazil. Roots are considered as the world's greatest supplier and β-ecdysone is the most important compound extracted from roots of Pfaffia glomerata. So, the aim this study was analyze the presence of β-ecdysone in the inflorescences and stems and compared with the content from roots of Pfaffia glomerata and determine the best extractive method of β-ecdysone this plant. The crude extracts were obtained by Soxhlet method, reflux, maceration, percolation and turbolyse. Compound extracts were quantified by High Performance Liquid Chromatography (HPLC). The analysis were carried out a Phenomenex Column C18, 5 µm, 250x4,6mm, maintened at 30 ºC, gradient system using as mobile phase a mixture of methanol and water, flow rate 1,0 mL and detection at 245 nm. Results showed Soxhlet method with ethanol:water (90:10 v/v) presented the higher concentration of β-ecdysone in P. glomerata and inflorescences showed higher amount of this active substance (3,06%), compared with stems (2,37%) and roots (1,63%), showing that the inflorescences and plant stems may also be used as a rich source of β-ecdysone.
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A família Amaranthaceae é formada por cerca de 2.360 espécies, 145 delas encontradas no Brasil. Cerca de 94 espécies da família subsistem em diversas fitofisionomias do Bioma Cerrado e 27 espécies aparecem em listas regionais de espécies ameaçadas de extinção. O objetivo deste trabalho foi inventariar e estudar a anatomia foliar e a morfologia de espécies da família Amaranthaceae de uma Unidade de Conservação de Alto Paraíso, GO, relacionando-as ao metabolismo fotossintético. Foram localizadas uma espécie de hábito subarbustivo (Pfaffia townsendii) e cinco espécies herbáceas (Froelichiella grisea, Gomphrena hermogenesii, G. lanigera, G. prostrata e P. gnaphalioides), a maioria demonstrando comportamento pirofítico e anemocoria, bem como sistemas subterrâneos bem desenvolvidos associados com anfiestomia foliar. A anatomia Kranz foi caracterizada em três espécies (todas do gênero Gomphrena), indicando o metabolismo fotossintético C4. Duas espécies são endêmicas da área e duas espécies são consideradas ameaçadas de extinção. Aspectos de anatomia e morfologia são discutidos em relação ao hábito das espécies, comportamento ecológico, duração das porções aéreas e significado funcional. Os dados demonstram a importância da família como indicadora da biodiversidade das áreas abertas dos cerrados e da importância da ampliação das pesquisas na Chapada dos Veadeiros, que tem potencial para o registro de novas espécies, inclusive endêmicas, dado o comportamento sazonal de algumas dicotiledôneas herbáceas e as dificuldades para localizá-las, identificá-las e coletá-las.
The Amaranthaceae family is composed of 2,360 species of which 145 are found in Brazilian vegetation. About 94 species of this family subsist in different phytofisionomies of the Cerrado Biome (a savanna-like vegetation) and 27 species are cited in Brazilian regional lists of endangered species. This work aimed to inventory and to study the leaf anatomy and morphology of the Amaranthaceae species found in one Conservation Area in Alto Paraíso, GO, relating them to the species' photosynthetic metabolism. It was found one subshrub species (Pfaffia townsendii) and five herbaceous species (Froelichiella grisea, Gomphrena hermogenesii, G. lanigera, G. prostrata and P. gnaphalioides), most of them showing pirophytic and anemocoric behavior and well developed subterraneous systems associated with leaf amphistomy. The Kranz anatomy was verified in three species (all Gomphrena genus), which indicates the C4 pathway of photosynthesis. Two species are endemic of the area (Chapada dos Veadeiros) and two are considered endangered species. The anatomy and morphology aspects are discussed in relation to the species habit, ecological behavior, life span of the aerial organs and functional data. The results indicate the Amaranthaceae importance as biodiversity indicator of open vegetation areas of Cerrado and the necessity of further research in Chapada dos Veadeiros, which has the potential to register new plant species, including endemic ones, since this work displays the seasonal behavior of some dicotyledonean herbaceous species and the difficulties to locate, identify and collect them.
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This study aimed to investigate the effects of the administration of butanolic residue (BR) of Pfaffia paniculata by intraperitoneal route to Ehrlich ascitis tumor bearing mice. Initially, a toxicity study of P. paniculata BR was performed in which doses of 12.5; 25 and 50mg/Kg were administered by intraperitoneal injection for seven days to Swiss mice. The treatment did not show toxicity. Then, Swiss male mice received, by intraperitoneal injection, once a day, 12.5; 25 or 50mg/Kg of P. paniculata BR for seven days. This protocol started in the same day of tumor inoculation with 5X10(6) cells i.p. The treatment with butanolic residue of P.paniculata i.p caused a significant increase in the ascitic volume; however, a significant decrease in tumor cells number per ml (p<0.05) was observed in P. paniculata treated mice that was followed by a numerical (although non-significant) decrease in the total numbers of tumor cells in the collected ascitic fluid. These results indicated a tumor cell inhibitory effect by P. paniculata butanolic residue in this experimental system, and indicate that topical application of this residue can be useful to control the cancer growth.
Neste estudo, foi avaliado o efeito do tratamento intraperitoneal com Resíduo Butanólico de Pfaffia paniculata, sobre o crescimento do Tumor de Ehrlich, forma ascítica. Foram utilizados dois grupos de 15 camundongos cada, sendo um grupo controle e o outro grupo tratado com RB 50mg/Kg. Todos os animais foram inoculados intraperitonealmente, com 5X10(6) células tumorais O tratamento iniciou-se no mesmo dia da inoculação do tumor. Assim, os animais receberam diariamente, por via intraperitoneal, 0,1 ml de RB na concentrações 50 mg/Kg, ou PBS como controle. Após 7 dias da inoculação do tumor, os animais foram eutanasiados e foi colhido o fluído ascítico total, para a contagem do número de células tumorais presentes neste fluído e estudo da morfologia destas células . Neste experimento observou-se aumento significante da quantidade de fluido ascítico nos animais tratados com RB, e diminuição significativa em relação ao número de células tumorais/ml e células tumorais totais, presentes no fluído ascítico, comparativamente com os animais controle. Estes resultados sugerem efeito inibitório tópico do RB levando à morte as células neoplásicas.
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Este estudo teve como objetivo otimizar a extração de ecdisterona em raízes de ginseng brasileiro. Primeiramente, para se avaliar a eficiência do solvente extrator, amostras de raízes dois acessos (BRA e JB-UFSM) de P. glomerata foram extraídas em Soxhlet com metanol e clorofórmio, separadamente, durante 4 horas. No segundo ensaio, com o intuito de se escolher o método extrator, a extração foi conduzida em Soxhlet e em ultrassom, utilizando metanol como solvente. Em P. tuberosa, as amostras foram extraídas com metanol, e a extração foi conduzida em Soxhlet e em banho ultrasônico. O conteúdo de ecdisterona foi determinado em Cromatógrafo Líquido de Alta Eficiência (CLAE). Em ambas as espécies, um maior conteúdo de ecdisterona foi detectado nas amostras extraídas com metanol e em Soxhlet. A metodologia proposta mostrou-se eficaz para a quantificação da ecdisterona a partir das raízes de P. glomerata e P. tuberosa, podendo ser aplicada no controle de qualidade de drogas vegetais e/ou fitoterápicos.
This study aimed at optimizing the extraction method from ecdysterone of Brazilian ginseng. Root samples of two accessions (BRA and JB-UFSM) of P. glomerata were extracted in a Soxhlet with methanol or chloroform for 4h. In the second trial, the extration was conduced in a Soxhlet or ultrasonic using metanol as a solvent. In P. tuberosa, the roots samples were extracted with methanol in a Soxhlet or in ultrasonic. The ecdysterone content was determinated using high efficiency liquid chromatography methods. In both studied species, the highest ecdisterone content was detected from samples extracted in a Soxhlet and using methanol as a solvent. This extration method has been successfully applied for determination of ecdysterone content from roots of Brazilian ginseng, and could be useful for the quality control of drugs and pharmaceutical formulations.
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Purpose: Pfaffia paniculata, a plant known as Brazilian ginseng, has been claimed by some patients suffering from sickle cell disease to have beneficial effects. In order to examine this assertion, a powder extract was obtained from the roots of the plant. Pacients and methods. The extract was studied to verified the desickling properties in vitro. We studied the behavior of blood cells treated with Pfaffia paniculate extract in vitro through morpholofic analyses of blood cells incubated with paffia paniculate extract in vitro. Thirty Brazilian patients with sickle cell disease receiving capsules containing the powder extract of Pfaffia paniculata (500 mg each) every 8 hours or capsules containing placebo were followed up for three months. The number of erythrocytes, reticulocytes, sickle cells, and peripheral erythroblast, hemoglobin (HB), hematocrit (HT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC), were determined in peripheral blood immediately before and 2 and 3 months after the beginning of the treatment. Results: Administration of Brazilian ginseng powder extract to patients having sickle cell resulted in decrease in sickle cells, erythroblast and reticulocytes in blood as well as increase in hemoglobin and hematocrit levels. Conclusion: The Pfaffic extract exhibited desickling properties when incubated with blood cells from deasese sickle cell disease patients or blood cells treated with 2% sodium methabisulphite. The clinical findings showed that treatment also led to improvement of the sintoms and signs in these patients.
Subject(s)
Humans , Male , Female , Anemia, Sickle Cell/drug therapy , Phytotherapeutic Drugs , Panax , Hematologic DiseasesABSTRACT
O presente trabalho objetivou avaliar o efeito do pH do meio de cultivo sobre alguns parâmetros de crescimento da Pfaffia glomerata (Spreng.) Pedersen cultivada in vitro, bem como checar se o crescimento dos explantes altera o pH do meio ao longo do período de cultivo. Foram testados quatro tratamentos constituídos de distintos valores de pH (3,7; 5,0; 6,0 e 7,5) do meio de cultivo. O pH do meio de cultivo foi ajustado antes da inclusão do agar (6g L-1 - Merck) e da autoclavagem. Como fonte de explantes foram utilizadas segmentos nodais de plantas previamente estabelecidas in vitro em meio MS. Dos nove aos 15 dias após a inoculação (DAI) dos segmentos nodais, verificou-se maior número de raízes em pH 6,0 e o menor no pH 7,5. Aos 35 DAI, o comprimento da maior brotação e o número total de segmentos nodais por planta foram maiores em torno de pH 6,0. Aos 35 DAI, observou-se menor crescimento em biomassa de raízes em pH 3,7. Já a parte aérea apresentou menor biomassa em pH 7,5. Aos 35 DAI, a produção de matéria fresca e seca total da plântula foi maior em pH próximo a 6,0. Concluiu-se que valores de pH do meio de cultivo próximos a 6,0, ajustados antes da autoclavagem, são ideais para o crescimento da P. glomerata cultivada in vitro. Também se verificou que o crescimento da plântula modificou significativamente o pH do meio de cultivo.
The present research aimed to evaluate the effect of culture medium pH on some growth parameter of Pfaffia glomerata (Spreng.) Pedersen in vitro cultured plantlets, as well as to check whether the explant´s growth alters the culture medium pH. Four treatments consisted of different values (3.7; 5.0; 6.0 and 7.5) of culture medium pH were tested. The culture medium pH was adjusted prior to the addition of agar (6g L-1 - Merck) and autoclaving. Nodal segments from asseptic plants grown in MS medium were used as explants. From 9 to 15 days after inoculation (DAI) of nodal segments, the higher number of roots was obtained at pH 6.0, and the lower at pH 7.5. At 35 DAI, both length of the higher sprout and total number of nodal segments per plantlet were greater at about pH 6.0. At 35 DAI, roots biomass was lower at pH 3.7. On the other hand, shoots biomass was lower at pH 7.5. Fresh and dry matter of the whole plantlet was greater at pH around 6.0. In conclusion, values of culture medium pH near to 6.0, adjusted prior to autoclaving, are ideal for the growth of P. glomerata in vitro cultured plantlets. Moreover, the in vitro growth of plantlet modified significantly the culture medium pH.
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A aquisição de nitrato de potássio PA, para qualquer finalidade, depende de autorização do Ministério da Defesa, o que resulta em dificuldades na sua aquisição. Com a finalidade de estudar a possibilidade da substituição desse reagente por outro produto encontrado livremente no comércio, foi conduzido um experimento onde se testaram várias concentrações de salitre potássico em comparação com o nitrato de potássio PA, como componente dos sais de MS. Utilizou-se o produto granulado MURER® nas seguintes concentrações (g L-1): a) 7,0; b) 7,4; c) 7,8; d) 8,2. Foi adicionado um tratamento-controle, com o KNO3 PA, conforme aparece nos sais de MS (1,9 g L-1), sendo a espécie testada à fáfia, a partir de segmentos nodais. O meio de cultura constituiu-se dos sais de MS, vitaminas de White, 30 g L-1 sacarose, 2,0 mg L-1 ácido indol butírico, 2,0 mg L-1 ácido naftalenoacético, 2,0 mg L-1 benzilaminopurina, pH 6,0 + 0,1 e 1,5 g L-1 Phytagel® como gelificante. O número médio de ramos formados para todos os tratamentos foi inferior ao do tratamento-controle; mas, em compensação, o comprimento médio deles foi superior, também em todos os tratamentos, sendo que o comprimento total de ramos na menor concentração de salitre, atingiu 17,0 cm, enquanto no tratamento-controle foi de 12,6 cm. A média da biomassa seca foi de 68,3 mg para a menor concentração de salitre e de 50,0 mg para o tratamento-controle. A análise dos resultados indicou ser vantajosa a substituição do nitrato de potássio PA pelo salitre potássico.
The acquisition of potassium nitrate for any purpose, including preparation of nutritive media for micropropagation, depends on authorization from the Ministry of Defense, what turns out in difficulties in its acquisition. With the purpose of studying the possibility of replacing this component of the culture for another product found freely in the market, an experiment was carried out, where several concentrations of potassic saltpetre were tried, in comparison to the chemically pure potassium nitrate, as a component of the MS salts formulation. The commercial granulated product MURER® was used in the following (g L-1): a) 7,0; b) 7,4; c) 7,8; d) 8,2; and e) chemically pure potassium nitrate (control-treatment) as it appears in the MS salts, or 1,9 g L-1Fáfia was tested, starting from nodal segments. The basal nutrient medium comprised the MS salts, White vitamins, 30 g L-1 sucrose, 2.0 mg L-1 indol butyric acid, 2.0 mg L-1 naftaleneacetic acid, 2.0 mg L-1 benzilaminopurine, pH 6.0 + 0.1 and 1.5 g L-1 Phytagel® as the gelling agent. The number of branches formed in all concentrations of potassic saltpetre was lower than those ones in the control treatment; however, their lengths were higher. The mean length of branches in the treatment with the smallest potassic saltpetre concentration, reached 17.0 cm in length, while in the control-treatment it was 12,6 cm. The dry biomass weight reached 68.3 mg for the smallest potassic saltpetre concentration, but only 50,0 mg for the control treatment. The results indicated to be advantageous the substitution of the chemically pure potassium nitrate for the potassic saltpeter, when either shoot elongation or biomass production is desired, being the nutrient medium sterilized with sodium hipoclorite. However, when a faster multiplication rate is the main objective, the nutrient medium should be prepared with chemically pure potassium nitrate.
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The present study was carried out to evaluate the acute toxicity and the effect of the aqueous extract of the roots from Pfaffia glomerata (Spreng) Pedersen (Amaranthaceae) (AEP) on the prevention of acetic acid-induced ulcer and on the healing process of previously induced ulcers. The acute toxicity was evaluated in Swiss mice after oral administration of a single dose and the chronic gastric ulcer was induced with local application of acetic acid. The results showed that the LD50 of the extract was 684.6 mg.kg-1 for the intraperitoneal administration and higher than 10 mg.kg-1by the oral route. The administration of the AEP did not prevent ulcers formation. However, the AEP increased of the healing process of previously induced ulcers. The results suggest that AEP chronically administered promote an increase of tissue healing, after the damage induced by acetic acid and the extract seemed to be destituted of toxic effects in the mice by the oral route.
Pfaffia glomerata (Spreng) Pedersen (Amaranthaceae), uma planta conhecida popularmente como "Ginseng Brasileiro" e "paratudo", é utilizada para tratar distúrbios gástricos e como cicatrizante. Em estudos anteriores, foi demonstrado que o extrato aquoso bruto da P. glomerata (AEP) protegeu a mucosa gástrica contra úlceras induzidas por etanol e estresse e reduziu a secreção ácida gástrica basal e estimulada em ratos com ligadura de piloro. Além disso, a secreção gástrica de animais tratados com AEP apresentou níveis de nitrato e nitrito aumentados. O objetivo deste estudo foi avaliar se o AEP previne o desenvolvimento de úlceras induzidas por ácido acético e o efeito desse extrato no processo de cicatrização em úlceras previamente formadas. A administração do AEP em diferentes doses produziu efeitos tóxicos baixos e não preveniu a formação de úlceras, porém aumentou o processo de cicatrização em úlceras já existentes, como evidenciado no estudo histopatológico. Em conclusão, o AEP administrado cronicamente promove o aumento da cicatrização do tecido após a lesão induzida com o ácido acético.
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Neste trabalho foi avaliado, em roedores, o efeito depressor das frações clorofórmio (CHCl3), acetato de etila (EtOAc) e n-butanol, obtidas das partes subterrâneas de Pfaffia glomerata, empregando-se o teste de tempo de sono barbitúrico como referência. Somente a fração lipofílica (CHCl3:EtOAc, 1:1, m/m) (i.p. 500 mg/kg; v.o. 1000 mg/kg) potenciou o tempo de sono induzido por pentobarbital. A ecdisterona foi isolada e identificada como constituinte majoritário (1,4 por cento m/m) desta fração, através de cromatografia líquida de alta eficiência e métodos espectroscópicos, respectivamente. Este composto potenciou o tempo de sono barbitúrico (100 mg/kg, i.p.; 400 mg/kg, v.o), sem causar hipotermia. Nestas mesmas doses, a ecdisterona não alterou a performance dos animais no rota-rod, esquiva inibitória e labirinto em cruz-elevado, além de não alterar o padrão de convulsões induzidas por pentilenotetrazol. Este perfil indica que esta substância, nestas doses, não apresenta perfil ansiolítico ou neurotóxico. Estes resultados indicam que a ecdisterona é o componente responsável pela ação hipnótica apresentada pela fração lipofílica obtida das partes subterrâneas de P. glomerata.
In this study the depressant effect of fractions from P. glomerata was initially evaluated using the mice barbiturate sleeping time test as reference. The fractions tested were the CHCl3, the EtOAc, the n-BuOH and the aqueous fraction obtained from P. glomerata subterraneous parts. Only the pretreatment with the lipophilic fraction (CHCl3: EtOAc, 1:1, w/w) increased the barbiturate sleeping time (i.p 500 mg/kg; v.o. 1000 mg/kg). Ecdysterone, the main substance isolated from this lipophilic fraction, was identified by spectroscopic methods and its content in the ethanol extract was determined as 1.4 percent (w/w) by HPLC. In order to investigate the hypothesis of ecdysterone displaying a depressant effect on nervous central system, an evaluation toward the hypnotic-sedative and anxiolytic effects of this drug was carried out. Ecdysterone 100 mg/kg, i.p, increased the barbiturate sleeping time without provoking hypothermia; when administered by oral route its minimal effective dose was 400 mg/kg. On the other hand, ecdysterone (100 mg/kg, i.p; 400 mg/kg, p.o) did not impair motor coordination and was ineffective on pentylenetetrazole-induced convulsion, elevated plus-maze and step-down inhibitory avoidance tests, indicating that at these doses the drug does not present an anxiolytic profile and does not cause manifest neurotoxic effects as well. In conclusion, the lipophilic fraction from P. glomerata presents a hypnotic effect being ecdysterone one of the compounds responsible for this CNS activity.
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Plants have been used for the human beings as food, as additives and/or as medicines. There are controversies about the biological effects of several natural products and, it is worthwhile to try to develop experimental assays to evaluate properties of extracts of plants. Pfaffia sp. is utilized in popular medicine and various properties have been attributed to its extract. Red blood cells (RBC) and plasma proteins are labeled with technetium-99m (Tc-99m) and this labeling procedure depends on a reducing agent and stannous ion is usually used. There are reports that drugs can alter the labeling of blood elements with Tc-99m. We have evaluated the influence of a Pfaffia sp. extract on the labeling of blood constituents with Tc-99m and on the morphology of RBC. Blood was incubated with an aqueous extract of Pfaffia sp., stannous chloride and Tc-99m. Samples were centrifuged and plasma and blood cells were separated and also precipitated with trichloroacetic acid. Soluble and insoluble fractions were separated. The results did not show alteration in the uptake of radioactivity and no modifications on the shape of the RBC in presence of Pfaffia sp. Once this labeling process depends on a reducing agent, probably, this extract has compounds with anti-oxidant properties as already described elsewhere, that could protect the stannous ions against the oxidation process. This fact would aid the labeling process of blood elements with Tc-99m.
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Analisou-se a variação sazonal e a influência de diferentes tipos de secagem (sol, sombra e estufa) no teor de saponinas e nas características físico-químicas da droga vegetal - raízes de Pfaffia glomerata (Spreng.) Pedersen (Amaranthaceae). A avaliação sazonal apontou a ocorrência de diferenças significativas nos teores de extrativos (maior no inverno: 61,1 por cento±1,9), saponinas totais (maior no inverno: 17,1±0,2 por cento), e entre cinzas totais (maiores no inverno e primavera: 6,5±0,4 e 6,5±1,0 por cento, respectivamente) e cinzas insolúveis em ácido(maiores no verão e outono: 0,57±0,11 e 0,44±0,18 por cento, repectivamente). Quanto à variação sazonal do teor de saponinas, o valor do lote colhido no inverno foi maior do que nas outras estações, confirmando o conceito geral de maior ocorrência de ativos em órgãos subterrâneos no inverno. Quanto à influência dos métodos de secagem, o lote seco ao sol forneceu maior teor de extrativos (61,2±2,3 por cento) do que os lotes secos na sombra e em estufa, contradizendo a regra geral de que a radiação solar degrada os ativos vegetais. Esse mesmo perfil de dados ocorreu com relação ao teor de saponinas, pois o lote seco ao sol mostrou maior valor(15,4 por cento) do que daqueles secos na sombra ou em estufa (14,4 e 11,6 por cento, respectivamente). Desta forma, verificou-se que, de fato, ocorre variação sazonal nas raízes de P. glomerata, particularmente, no que diz respeito aos teoes de extrativos, cinzas e saponinas totais, sendo que essas variações indicam maior concentração de ativos nos períodos de inverno e primavera. Dentre os métodos de secagem testados, o emprego do sol parece favorecer os teores de saponinas e de extrativos, embora tal conclusão mereça ser melhor investigada.
The seasonal variation and the influence of drying (sun, shadow and hot-house greenhouse) were analysed in relation to the total saponin content and physicochemical characteristics. The seasonal evaluation detected significant differences in the parameters of extractable matter (larger in winter: 61.1 percent±1.9), total saponin content (larger in winter; 17.1±0.2 percent) and between total ash (larger in winter and spring; 6.5±0.4 and 6.5±1.0 percent, respectively) and acid-insoluble ash (larger in summer and autumn; 0.57±0.11 and 0.44±0.18 percent, respectively). In relation to the seasonal variation of total saponin content, the value of the sample collected in winter was larger than in other seasons, confirming the general rule that the largest content of substances in underground organs occurred in winter. In relation to the influence of drying methods, the sample dried at the sun presented a larger value of extractable matter (61.2±2.3 percent) than those dried on shadow and in hot-house greenhouse, contradicting the general rule that the sun radiation would degrade natural substances. The same profile was found in relation to the total saponin content, because the sample dried at the sun showed a larger value (15.4 percent) than those dried on shadow or in hot-house (14.4 and 11.6 percent, respectively). However, these conclusions have to be more investigated.
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Os relatos encontrados na literatura científica sobre a composição química e atividades farmacológicas do gênero Pfaffia spp. são apresentados. Os estudos sobre esse gênero são ainda bastante escassos, sendo assim, o uso racional de espécies de Pfaffia, com finalidade terapêutica ainda depende de conhecimento aprofundado de suas propriedades farmacológicas e do desenvolvimento de tecnologias de produção e controle de qualidade.
This review summarizes available chemical and pharmacological data about Pfaffia genre. Scientific studies about these species are still lacking, and their therapeutical rational use depends on the availability of more informations about their pharmacological properties and the development of production and control quality technologies.
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As raízes secas da espécie brasileira Pfaffia tuberosa (Spreng.) Hicken foram submetidas a extração com metanol. Deste extrato metanólico foram isolados a ecdisterona e o ácido oleanólico, através de processos de separação por HPLC e cromatografia em coluna. Tanto a ecdisterona como o ácido oleanólico foram identificados comparativamenve com os respectivos padrões. Análise quantitativa por HPLC mostrou 0,23 por cento de ecdisterona. Este teor foi considerado baixo em relação a outra espécie, também brasileira, a Pfaffia iresinoides.
In this paper, we wish to report the isolation and identification of ecdysterone from Pfaffia tuberosa (Spreng.) Hicken. The roots of Pfaffia tuberosa (Spreng.) Hicken were colected in the Mato Grosso State - Brazil. The dried and sliced roots (197g) were extracted with hot MeOH. A suspension of the resulting MeOH extract in the water was treated with n-BuOH. The n-BuOH layer was concentrated in vacuo to give a brown viscous oil (5g), wich was chromatographed on silicagel (65g) using CHCl3 - MeOH - H2O (80:20:10) lower phase, and 15ml fractions were collected. Fractions 11-13 were concentrated and the residual solid was recrystallized from EtOH to afford oleanolic acid (65mg). The crystalline residue obtained from fractions 44-56 was recrystallized from EtOAC-MeOH to give ecdysterone (87mg). Each compound was identified by direct comparison with an authentic sample. Further HPLC quantitative analysis showed lower content of ecdysterone (0,23 percent) compared with that of Pfaffia iresinoides Spreng.