Authentication of Brazilian Ginseng using Bar-HRM analysis

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".

In vitro and in vivo antioxidant potentials of the methanolic crude extract from Inula glomerata Oliv. & Hiern (Asteraceae) and Salacia kraussii (Harv) Harv (Celastraceae) / Potenciales antioxidantes in vitro e in vivo del extracto crudo metanólico de Inula glomerata Oliv. & Hiern. (Asteraceae) y Salacia kraussii (Harv) Harv (Celastraceae)

Reactive oxygen species are implicated in multiple pathological conditions including erectile dysfunction. This study evaluated the in vitro and in vivo antioxidant potential of the methanolic extracts of Inula glomerata and Salacia kraussii. The plant materials were pulverized and extracted with methanol. The phytochemical analysis, ability of the crude extracts to scavenge free radicals (ABTS, DPPH, NO.) in vitroas well as the total phenolic and flavonoid contents was investigated. In vivo, antioxidant potentials of the crude extracts (50/250 mg/kg body weight) were determined in an erectile dysfunction rat model. The phytochemical analysis revealed that both plants contain flavonoids, tannins, terpenoids, and alkaloids. The crude extracts at varying degree of efficiency, scavenged ABTS and DPPH radicals. The crude extracts at low concentrations (50 mg/kg b.w) significantly (p<0.05) diminished the level of malondialdehyde, augmented catalase activities and elevated glutathione levels. However, SOD activities were significantly boosted in a dose-dependent manner by the crude extracts. Therefore, I. glomerataand S. kraussiipossess antioxidant properties, hence, can serve as a therapeutic modality in the treatment of oxidative stress-induced erectile dysfunction.

Las especies reactivas de oxígeno están implicadas en múltiples condiciones patológicas, incluyendo la disfunción eréctil. Este estudio evaluó el potencial antioxidante in vitro e in vivo de extractos metanólicos de Inula glomeratay Salacia kraussii. Los materiales vegetales fueron pulverizados y extraídos con metanol. A estos extractos crudos se les llevó a cabo el análisis fitoquímico junto con el contenido total de fenólicos y flavonoides, así como se les investigó la capacidad in vitro para atrapar radicales (ABTS, DPPH, NO.). Los potenciales antioxidantes in vivo de los extractos crudos (50/250 mg/kg de peso corporal) se determinaron en un modelo en ratas con disfunción eréctil. El análisis fitoquímico reveló que ambas plantas contuvieron flavonoides, taninos, terpenoides y alcaloides. Los extractos crudos con un grado variable de eficiencia, atraparon a los radicales ABTS y DPPH. Los extractos crudos a bajas concentraciones (50 mg/kg p.c) significativamente (p<0.05) disminuyeron el nivel de malondialdehído, aumentaron las actividades de catalasa y elevaron los niveles de glutatión. Sin embargo, las actividades de SOD por los extractos crudos fueron significativamente dosis-dependientes. Así, los extractos de I. glomeratay S. kraussii mostraron propiedades antioxidantes, y por lo tanto, podrían servir como una alternativa terapéutica en el tratamiento de disfunción eréctil inducida por estrés oxidativo.

β-ecdysone content and antioxidant capacity in different organs of Brazilian ginseng / Teor de β-ecdisona e capacidade antioxidante em diferentes órgãos de ginseng brasileiro

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.

Stability of hydroalcoholic extracts of two species of guaco; Mikania glomerata SPRENG. and Mikania laevigata SCHULTZ. (Asteraceae), by UHPLC-MS

It is important to study the stability of plant extracts used as active ingredients in phytotherapic medicine, as degradation of the active principles directly affects the efficacy and safety of these products. Therefore, a stability study of the hydroalcoholic extract of the species: Mikania glomerata and Mikania laevigata was conducted in order to determine the speed of degradation and shelf life of these extracts, which are incorporated in cough syrup in Brazil. Leaves of both species were dried in an oven or by lyophilization (freeze-dried). Hydroalcoholic extracts underwent both accelerated stability study of six months and long-term stability study for 12 months. Samples were stored at different temperatures and every three months were analysed by ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS) to monitor their chemical profile, quantifying coumarin and chlorogenic acid. For all conditions of the study, a reduction of the content of the chemical marker of this species, coumarin, greater than 5% was observed, so a shelf life of two years cannot be assigned to the hydroalcoholic extracts of these species as observed in commercial extracts.

Teratogenic effects of aqueous extract of Ficus glomerata leaf during embryonic development in zebrafish (Danio rerio)

Over the past few decades, many reports were published in scientific journals describing medicinal properties of Ficusglomerata (FG). However, its effects on embryonic development and its safety characteristics have not been studied.The purpose of this investigation was to determine lethal concentration 50 (LC50) and study the effect of aqueousextract of FG leaf (AEFG) on developmental abnormalities in zebrafish embryos. LC50 value of AEFG was calculatedby using probit analysis. Effect on percentage hatchability, heartbeat rate, total body length, and developmentalmorphological abnormalities, i.e., delayed growth, abnormal movement, tail detachment, abnormal head-trunkangle, scoliosis/flexure, and yolk sac edema were recorded. AEFG revealed LC50 of 239.88 ppm. The result showeda significant reduction in percent hatchability (p < 0.05), heartbeat rate (p < 0.001), total body length (p < 0.001),and developmental morphological abnormalities in the embryos treated with AEFG. This research can be used inconsidering the safety of an AEFG extract for their use during pre-conception or early pregnancy period.

Preliminary phytochemical screening and GC-MS analysis of Cladophora glomerata: green marine algae

Background: Seaweeds since ages are excellent source of biologically active ingredients. Several Asian countries have a strong tradition of using various seaweeds in herbal medicines preparations. These plants contain various phytochemical constituents having biological activities. Seaweeds are the source of phytochemicals namely agar-agar, carrageenan and algin, which are extensively used in various industries such as food, confectionary, textiles, pharmaceuticals, dairy and paper industries mostly as gelling, stabilizing and thickening agents. They are also used for human consumption, animal feed and as manure in several countries. Several Asian countries are using various seaweeds in traditional medicines.Methods: In the present study, the shade dried and methanolic extract of Cladophora glomerata, a marine green algae was subjected to preliminary phytochemical and gas chromatography-mass spectrometry analysis(GC-MS) to identify the various bioactive components.Results: The methanolic extract of Cladophora glomerata revealed the presence of alkaloids, glycosides, flavonoids, saponins, diterpenes and carbohydrates. The GC-MS analysis of the methanolic extract of Cladophora glomerata showed the presence of 42 different compounds. The major compounds were dibutyl phthalate (27.07%), hexadecanoic acid, methyl ester (9.58%), 1,2-benzene-di-carboxylic acid (8.11%), octatriacontyl trifluoroacetate (6.81%), cholesterol (6.66%).Conclusions: Thus, in the present study of Cladophora glomerata, phytochemical and GC-MS analysis provides an important novel information to support further ongoing studies to evaluate structure of bioactive compound and its pharmacological activities.

Radiation levels of UV-A and UV-B on growth parameters and coumarin content in guaco / Níveis de radiação UVA e UVB sobre parâmetros de crescimento e conteúdo de cumarina em guaco

ABSTRACT: Guaco is a medicinal plant species containing coumarin, a secondary metabolite whose concentration is influenced by various factors, such as the level of exposure to UVA (315-400 nm) and UVB (280-315 nm) radiation, age of the plant, and position of the storage organ in the plant. This study aimed to assess the effect of different blocking levels of UVA and UVB radiation on growth parameters and coumarin content of two guaco accessions. Treatments did not affect any of the growth parameters assessed, except leaf area. Coumarin content was higher in leaves collected from the upper canopy and with unblocked exposure to UVA and UVB.

RESUMO: Guaco é uma espécie medicinal que apresenta em sua composição a cumarina, um metabólito secundário cuja concentração no vegetal é influenciada por fatores como os níveis de radiação UV-A (315-400 nm) e UV-B (280-315 nm) incidentes, bem como pela idade e posição do órgão de armazenamento na planta. O objetivo deste estudo foi o de avaliar o efeito de diferentes níveis de bloqueio das radiações UV-A e UV-B sobre parâmetros de crescimento e teor de cumarina em dois acessos de guaco. Os tratamentos não apresentaram efeito sobre os parâmetros de crescimento avaliados, com exceção dos valores de área foliar. Em relação aos teores de cumarina, estes foram superiores nas folhas coletadas do dossel superior, bem como no tratamento sem bloqueio das radiações UV-A e UV-B incidentes.

Comparison of oleanolic acid content in Pfaffia glomerata from different habitats of Guangxi / 国际药学研究杂志

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.

Optimization of the extraction and enrichment process and evatuation of the free radical scavenging activity of triterpene sapogenins from Pfaffia glomerata / 国际药学研究杂志

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.

Chemical constituents from ethyl acetate extracts of domestic Pfaffia glomerata / 中草药

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.

UHPLC–MS quantification of coumarin and chlorogenic acid in extracts of the medicinal plants known as guaco (Mikania glomerata and Mikania laevigata)

Abstract In Brazil, Mikania glomerata Spreng. and M. laevigata Sch. Bip. ex Baker, Asteraceae, known popularly as guaco, are widely used for colds and asthma. Although coumarin is adopted as the chemical marker of both species, it was not always detected in M. glomerata, for which chlorogenic acid was identified and quantified instead. The purpose of this study was to develop and validate a method to quantify both coumarin and chlorogenic acid and apply it to extracts of plants identified as M. glomerata, M. laevigata, or as guaco, to determine the pattern of composition of these two species and to observe differences between oven-dried and lyophilized leaves. A method using ultra-high resolution liquid chromatography–mass spectrometry (UHPLC–MS) in the full scan mode was validated for selectivity, matrix effect, linearity, limits of detection and quantification, precision and accuracy. The concentration of coumarin varied between species and samples, therefore these two species should not be used interchangeably. The concentration of chlorogenic acid was also determined for all samples. The UHPLC–MS method permitted the quantification of coumarin and chlorogenic acid in 16 samples of guaco and several commercial samples were possibly misidentified.

Quantificação de citocinas no soro e homogenato da pata na intoxicação experimental com veneno de Bothropoides jararaca em ratos Wistar tratados com soroterapia e Mikania glomerata / Quantification of cytokines in serum and paw homogenate of experimental intoxication for venom of the Bothropoides jararaca in Wistar rats treated with antivenom and Mikania glomerata

O presente estudo teve como objetivo quantificar os níveis de citocinas pró-inflamatórias, entre as quais TNF-α, interleucina-1β (IL-1β), IL-6, e anti-inflamatórias, como IL-10, interferon-γ (INF-γ), bem como comparar o efeito do tratamento convencional com o efeito do tratamento complementado pelo extrato da planta Mikania glomerata, na intoxicação experimental por Bothropoides jararaca. Foram usados ratos Wistar,divididos em três grupos: C - controle, VB - veneno botrópico + soro antiofídico e VBM - veneno botrópico + soro antiofídico + Mikania glomerata. As citocinas foram quantificadas, no soro e no homogenato desses animais, pelo teste ELISA, em três momentos (M1 - 30 minutos, M2 - seis horas e M3 - 24 horas após a inoculação do veneno). Os resultados obtidos evidenciaram que a intoxicação por veneno botrópico estimula principalmente a produção de IL-6 no soro e TNF-α, IL-1β, IL-6 no homogenato da pata de animais experimentalmente intoxicados. O tratamento complementar, com o extrato da planta Mikania glomerata, teve influência principalmente na produção de IL-6, IL-10 e IFN-γ no soro e IL-6, IL-1β e IFN-γ no homogenato. Porém, são necessários novos estudos com o extrato de Mikania glomerata para que se possa entender a ação dessa planta sobre a intoxicação botrópica, bem como verificar qual a melhor via para administrá-lo...

This experiment aimed to quantify the pro-inflammatory cytokine levels, including TNF-α, interleukin-1β (IL-1β) and IL-6 as well as the anti-inflammatory ones such as IL-10 and INF-γ. It was also proposed to compare the effect of the conventional treatment to a treatment in which was added the Mikania glomerata plant in the experimental intoxication using Bothropoides jararaca venom. It was used Wistar rats that were randomly divided into 3 groups: C - control; VB - Bothrops venom + antivenom serum; and VBM - Bothrops venom + antivenom serum + Mikania glomerata. Cytokines were quantified in the serum and paw homogenate using ELISA test in three different moments (M1- 30 minutes, M2- 6 hours and M3- 24 hours after venom injection). The intoxication by Bothropoides jararaca venoms mainly stimulated the production of IL-6 in the serum and TNF-α, IL-1β, IL-6 in paw homogenate of animals experimentally intoxicated. Adjunctive treatment with the extract of the Mikania glomerata plant mainly influenced the production of IL-6, IL-10 and IFN-γ in the serum and IL-6, IL1β and IFN-γ in paw homogenate. Further research is necessary with the extract of Mikania glomerata in order to understand the action of this plant on the Bothropoides poisoning and also to verify the best way to manage it...

Analgesic and Anti-inflammatory Effect of the Aqueous Extract of Dichrostachys glomerata (Forssk.) Hutch Fruits.

Aims: Dichrostachys glomerata is a plant found in the humid areas of Africa and widely used for the treatment of many ailments including rheumatism and snake bite. The present study has been undertaken to assess the analgesic and the anti-inflammatory properties of aqueous extract Dichrostachys glomerata fruit. Place and Duration of Study: Department of Animal Biology and Physiology, Faculty of Science, University of Yaounde I, Cameroon. Between January 2012 and October 2012. Methodology: Pain was induced in mice by the intraperitoneal administration of 1% acetic acid, hot plate, formalin and tail immersion test. Carrageenan and serotonin (1%) were used to induce inflammation in rat paws. Results: Dichrostachys glomerata significantly inhibited pain induced by acetic acid with a percentage inhibition of 19.4, 69.8, 33.7 and 24.3% respectively at the doses of 25, 50, 100 and 200 mg/kg. An acute pretreatment of mice with extract significantly increased reaction time in the hot plate test with a percentage inhibition more than 68%. Formalin induced pain was also significantly inhibited after treatment of rat with the plant extract at the doses of 25 and 50 mg/kg for the neurogenic phase with percentage of inhibition of 56.14 and 61.46% respectively. The extract significantly reduced oedema induced by carrageenan injection with a PI of 72.57 and 79.85% at the doses of 200 and 25 mg/kg respectively. In contrast, a pi of 65.03% was obtained with the plant extract at the dose of 50 mg/kg on serotonin-induced oedema. Conclusion: The Results obtained showed that D. glomerata aqueous extract have both analgesic and anti-inflammatory properties and could be a potential source of new oral anti-inflammatory and/ or analgesic drug.

Estudo das propriedades físico-químicas e avaliação da toxicidade aguda do extrato etanólico das folhas de Mikania glomerata Sprengel / Study of the physicochemical properties and acute toxicity evaluation of the ethanolic extract of the leaves of Mikania glomerata Sprengel

A Mikania glomerata é uma planta pertencente à família Asteraceae que é bastante utilizada na medicina popular devido às suas ações broncodilatadora, antiasmática, expectorante e antitussígena. O objetivo do presente estudo foi determinar as propriedades físico-químicas do pó obtido a partir das folhas de M. glomerata, bem como, avaliar a toxicidade em camundongos após tratamento agudo com doses repetidas do extrato etanólico padronizado preparado a partir das folhas dessa espécie. Durante o estudo das propriedades físico-químicas do pó obtido a partir das folhas de M. glomerata foram feitas as determinações da densidade bruta e de compactação, do teor de cinzas totais, do teor de umidade, e da granulometria. De acordo com os resultados obtidos podemos sugerir que o pó pode ser usado na formulação de uma forma farmacêutica sólida. Na segunda parte do estudo foi determinada a Dose Letal 50% (DL50), bem como, realizada a análise morfológica macroscópica e avaliados a toxicidade aguda com doses repetidas e os parâmetros bioquímicos e hematológicos de camundongos. De acordo com os dados obtidos na segunda parte deste estudo podemos sugerir que o extrato etanólico pode ser usado de forma segura em humanos, uma vez que apresentou valor de DL50 de aproximadamente 3000 mg Kg-1), bem como, não produziu nenhuma alteração morfológica nos principais órgãos, e nem provocou alterações nos parâmetros bioquímicos e hematológicos de camundongos.

The Mikania glomerata is a plant belonging to the Asteraceae that is widely used in folk medicine because of its bronchodilator, antiasthmatic, expectorant and antitussive actions. The aim of this study was to determine the physicochemical properties of the powder obtained from the leaves of M. glomerata, and to evaluate its toxicity in mice after acute treatment with repeated doses of standardized ethanol extract prepared from the leaves of this plant species. We determined the bulk and packing density, the total ash content, the moisture content and the particle size. The results suggest that the powder can be used in the formulating of a solid pharmaceutical form. In the second part of this study, we determined the 50% lethal dose (LD50), performed the gross morphological analysis and evaluated the acute toxicity from the use of repeated doses and the biochemical and hematologic parameters in mice. The data obtained in this part suggest that the ethanol extract can be used safely in humans, since it has a LD50 value of approximately 3000 mg kg -1 and produced no morphological changes in the major organs, or caused alterations in the biochemical and hematological parameters in mice.

A Case of Phoma glomerata Keratitis Occurred in Recurrent Herpes Simplex Keratitis Cicatrix

PURPOSE: To report a case of Phoma glomerata keratitis occurring in recurrent herpes simplex keratitis cicatrix. CASE SUMMARY: A 63-year-old male patient was admitted to our hospital with complaints of abrupt visual deterioration and ocular pain in his left eye. He was treated for recurrent herpes simplex keratitis in the same eye 12 years prior. Because central desmatocele was observed as a result of advanced corneal stromal melting, Gram staining, Potassium Hydroxide (KOH) mount, and culture were performed in corneal scrape specimens. On microbiological evaluation, a Phoma species was detected and Phoma glomerata was diagnosed using DNA sequencing method. Two consecutive amniotic membrane transplantations were performed with topical antifungal agents. The lesion was not improved when using topical amphotericin B and natamycin eyedrops, thus fluconazole eyedrops were used additionally. The corneal infection was resolved with central thick opacification. CONCLUSIONS: In the present case, herpetic keratitis was the main underlying causative factor because the patient had no past history of trauma. When diverse appearances of keratitis occur in herpes simplex keratitis patients, clinicians need to consider the concurrence of fungal infection, especially Phoma glomerata, a rare fungal organism.

Propriedades físicoquímicas e avaliação da toxicidade aguda do extrato etanólico padronizado a 70% das folhas de Mikania glomerata (Asteraceae) / Physicochemical properties and acute toxicity evaluation of ethanol extract of the leaves from Mikania glomerata Sprengel

Mikania glomerata (Asteraceae) é bastante utilizada na medicina popular devido às suas ações broncodilatadora, antiasmática, expectorante e antitussígena. O objetivo do presente estudo foi determinar propriedades físicoquímicas do pó obtido a partir das folhas de M. glomerata, bem como avaliar a toxicidade em camundongos após tratamento agudo com doses repetidas do extrato etanólico padronizado a 70% preparado durante 30 dias consecutivos. No estudo das propriedades físicoquímicas fez-se a determinação da densidade bruta e de compactação, do teor de cinzas totais, do teor de umidade e da granulometria. De acordo com os resultados obtidos o pó pode ser usado na formulação de uma forma farmacêutica sólida, uma vez que suas propriedades físico-químicas são compatíveis com o desenvolvimento desse tipo de formulação. Na segunda parte do estudo foi determinada a dose letal 50% (DL50) em camundongos, e na análise morfológica macroscópica dos principais órgãos e avaliada a toxicidade aguda com doses repetidas em parâmetros bioquímicos e hematológicos de camundongos. Os resultados sugerem que o extrato etanólico padronizado a 70% pode ser usado de forma segura, uma vez que apresentou um valor para a DL50 (~3000 mg kg-1) que pode ser classificado na categoria nociva, e não produziu nenhuma alteração morfológica nos principais órgãos e em parâmetros bioquímicos e hematológicos de camundongos.

Mikania glomerata Sprengel is a plant from the Asteraceae family and it is widely used in folk medicine because of its bronchodilating, anti-asthmatic, expectorant and antitussive effects. The purpose of this study was to determine the physicochemical properties of the powder obtained from the leaves of M. glomerata, and to evaluate the toxicity in mice after an acute treatment with repeated doses of standardized 70% ethanol extract prepared from the leaves during 30 consecutive days. To study the physicochemical properties of the powder, we conducted a determination of the bulk density and compaction, the total ash content, the moisture content and particle size. According to the results obtained, we suggest that the powder can be used to formulate a solid dosage form, since its physicochemical properties matches the development of this type of formulation. In the second part of the study, it was determined a lethal dose in the order of 50% (LD50), along with a gross morphological analysis and the evaluation of the acute toxicity with repeated doses, in the terms of biochemical and hematological parameters of mice. According to the results from the second phase, we suggest that the 70% ethanol extract can be used safely in humans, since it presented a value for the LD50 (~ 3000 mg kg-1) that can be classified as 'harmful'. It also did not produce any morphological changes in the major organs and in the biochemical and hematological parameters of mice.

Quantification of β-ecdysone in differents parts of Pfaffia glomerata by HPLC

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.

Rendimento extrativo de cumarina de folhas de guaco (Mikania glomerata Sprengel) submetidas a diferentes temperaturas de secagem / Extraction yield of coumarin from guaco (Mikania glomerata Sprengel) leaves subjected to different drying temperatures

O objetivo deste trabalho foi avaliar o efeito da temperatura do ar de secagem no rendimento extrativo da cumarina de folhas de guaco. Foram empregados 6 tratamentos de secagem, sendo ar ambiente, ar aquecido a 40, 50, 60, 70 e 80ºC. Utilizou-se secador de bandejas, tendo como fonte de aquecimento o gás liquefeito de petróleo (GLP). Os rendimentos extrativos da cumarina, depois de realizada a secagem, foram comparados com os valores obtidos da planta fresca (tratamento testemunha). A extração da cumarina foi realizada pelo método a quente, em banho-maria a 65ºC, sendo a identificação e quantificação realizada por cromatografia líquida de alta eficiência (CLAE). Em função dos resultados obtidos, pôde-se concluir que a temperatura do ar de secagem a 50ºC possibilitou o melhor resultado para o rendimento extrativo de cumarina em folhas de guaco.

The aim of this study was to evaluate the effect of drying on the extraction yield of coumarin from guaco leaves. Six drying treatments were used, being room air, heated air at 40, 50, 60, 70 and 80ºC. A tray dryer was used with liquefied petroleum gas (LPG) as heating source. The extraction yield of coumarin, after drying, was compared to the values obtained from the fresh plant (control treatment). Coumarin extraction was carried out by using the heat method, in water bath at 65ºC, and identification and quantification were done by means of high performance liquid chromatography (HPLC). Considering the obtained results, the temperature of the drying air at 50ºC led to the best result for the extraction yield of coumarin in guaco leaves.

First Record of Anastrepha flavipennis Greene (Diptera: Tephritidae) and of its Host in the Brazilian Amazon

Anastrepha flavipennis Greene was obtained from Pouteria glomerata (Sapotaceae) fruits, known as "abiurana-da-várzea" in the Brazilian Amazon. This is the first record of A. flavipennis for the state of Amazonas and of P. glomerata as a host for this fruit fly in the Amazon Basin.

Toxicological screening of Mikania glomerata Spreng., Asteraceae, extract in male Wistar rats reproductive system, sperm production and testosterone level after chronic treatment / Triagem toxocológica do extrato de Mikania glomerata Spreng., Asteraceae, no sistema reprodutor, produção espermática e nível de testosterona em machos de ratos Wistar após tratamento crônico

Some compounds present in therapeutic plants may be responsible for the occurrence of adverse side effects. Coumarin and flavonoids are substances found in many plant species that showed antifertility activity in female rats and dogs, respectively. Mikania glomerata Spreng., Asteraceae, known as guaco in Brazil, is a plant largely used in folk medicine and its leaves are reported to have coumarin and flavonoids. This work analyzes the effect of chronic administration of M. glomerata on the reproductive system of male rats. Thirty-day-old Wistar rats were treated with M. glomerata hydroalcoholic extract at a dose of 3.3 g/kg of body weight for ninety days. Body and organ weights, gamete concentration on the epididymis cauda, serum testosterone level and food consumption were evaluated. No significant alteration was observed in any of the variables analyzed, suggesting the absence of toxic action or antifertility activity of the M. glomerata hydroalcoholic extract.

As plantas medicinais podem apresentar na sua constituição compostos capazes de causar efeitos adversos no organismo. Cumarina e flavonoides são substâncias encontradas em muitas espécies vegetais, cuja interferência na fertilidade de ratas e cadelas, respectivamente, foi evidenciada em estudos prévios. Mikania glomerata Spreng., Asteraceae, (guaco) é uma planta usada no tratamento de doenças respiratórias e em suas folhas foi detectada a presença de cumarina e flavonoides. Neste estudo, avaliou-se o efeito do extrato hidroalcoólico, preparado com partes aéreas de guaco, no sistema reprodutor de ratos submetidos a tratamento crônico. Ratos Wistar (trinta dias de idade) foram tratados com extrato hidroalcoólico de guaco na dose de 3,3 g/kg de peso corporal durante noventa dias. O peso corporal e de órgãos, a produção de espermatozoides, a concentração de testosterona plasmática e o consumo de ração foram avaliados. Não foi observada nenhuma alteração significativa das variáveis analisadas e o tratamento não afetou o consumo de ração. Estes dados sugerem que, na dose utilizada, o extrato hidroalcoólico de guaco não teve efeito tóxico e nem interferiu com a fertilidade de ratos Wistar submetidos a um tratamento de longa duração.