Preparative separation of the naphthopyranone glycosides by high-speed counter-current chromatography

High-speed counter-current chromatography was applied to the preparative separation and purification of naphthopyranone glycosides from a crude 70% ethanolic extract of the capitula of Paepalanthus microphyllus. The solvent system used was composed of water-ethanol-ethyl acetate-hexane (10:4:10:4, v/v). This technique led to the separation of four different naphthopyranone glycosides in pure form in only 7 h. (C) 2001 Elsevier B.V. B.V. All rights reserved.

Planifolin, a new naphthopyranone dimer and flavonoids from Paepalanthus planifolius

A new naphthopyranone dimer (I) named planifolin was isolated from a methylene chloride extract of the capitula of Paepalanthus planifolius. The structure of 1 has been determined by chemical and spectroscopic means. In addition, a known dihydronaphthopyranone glycoside and seven known flavonoids were isolated from an ethanolic extract of the leaves of P. planifolius.

Estudo farmacognóstico do caule e raízes de Microgramma squamulosa (Kaulf.) Sota (Polypodiaceae)

Pharmacognostic and phytochemical study of the stem and root,,of Microgramma squamulosa (Kaulf.) Sota. Erva silvina, or Microgramma squamulosa (Kaulf.), Sota belongs to the family Polylodiaceae and is traditionally used as an antiulcer agent. Previous assays showed results that confirm the antiulcer action of the ethanolic extract and some fractions of the stem of M squamulosa and results showing no significant toxicity in the acute model, both made in rats Due to the significant results obtained in this previous work, it becomes important to furnish tools to identify the vegetal drug. The species is' an epiphyte that bears lanceolate scales totally covering the long-creeping brown-colored stem. Adventicious roots can be seen in the abaxial portion of, the-stem, while the leaves appear in the adaxial portion. Elements which contribute to the histological identification are: peltate lanceolate -haired scales, sclereids, meristeles (amphicribal bundles) and scalariform the roots. Phytochemical screening, and TLC tracheids in the stem; strip hairs and sclereids in analysis showed the presence of flavonoids and tannins which may be related to the anti,ulcer the plant as well as the characterization of activity. Results shown may help the identification of the fragmented vegetal drug and if it is presented as powder or as extract.

Atividade antimicrobiana de extratos brutos de Petiveria alliacea L.

"Antimicrobial activity of crude extratcs of Petiveria alliacea L.". Petiveria alliacea L. (Phytolaccacea) is an herbaceous plant of great importance in traditional medicine. This species have been widely used in several applications such as antirheumatic, anticarcinogenic, anti-flu, antitussive, analgesic, insecticidal, acaricidal, as well as bactericide and fungicide. Currently, the pathogenic microorganisms are acquiring resistence against the traditional antibiotics, and the search for new herbal antimicrobial agents has been intensified. The objective of this study was to evaluate the antifungal and antibacterial activity of several leaf crude extracts of P. alliacea against several strains of bacterias and yeasts namely Bacilus subtilis, Pseudomonas aeruginosa, Escherichia coli, Streptococcus mutans, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Candida parapsilosis, Candida kefyr and Candida albicans, using microdilution method. Promising results were observed for the 70% v/v ethanolic extract which presented minimum inhibitory concentration (MIC) from 250 to 760 mu g/mL for yeast. For the bacteria strains tested the MIC ranged between 240 to 3960 mu g/mL, depending of the extractive solution tested.

Naphthopyranone glycosides from Paepalanthus bromelioides

Two new naphtho[2,3-C]pyran-1-one glycosides, paepalantine-9-O-β-D- glucopyranoside and paepalantine-9-O-β-D-allopyranosyl(1 → 6)glucopyranoside, were isolated from an ethanolic extract of capitula from Paepalanthus bromelioides and identified from their spectrometric data.

Avaliação do extrato de Momordica charantia em Salmonella typhimurium: Mutagenicidade e antimutagenicidade

In the last years some natural products has been described as supressors of the mutagenic process in bacteria, the antimutagenics. The literature reference that in most of the countries, the population makes use of medicinal plants. The plant Momordica charantia (Cucurbitaceae) is original from Africa being used popularly as purgative, antirheumatic and for skin problems, burns and hemorrhoids. The present work had as objective to evaluate the mutagenic and antimutagenic activities of the ethanolic extract of M. charantia in Salmonella/microsome assays using TA100, TA98 and TA102 strains. It was verified that the extract did not present mutagenic activity when evaluated in different concentrations (0.64, 1.27, 2.55 and 3.84 mg/plate) but acted as antimutagenic agent against the mutations induced by the sodium azide (TA100,-S9), 4-nitro-phenylenediamine (TA98, -S9), daunomycin (TA102, +S9) 2-anthramine (TA100 and TA98, +S9) and 2-aminofluorene (TA102, +S9). When the metabolic activation (+S9) was used, the percentage of inhibition of the mutagenicity varied in the range of 31%-96%, while in absence of metabolizing system (-S9), the maximum percentage of inhibition of the mutagenicity was 44%. In that way, we can conclude that the metabolites found in the extract has potential to protect the genetic material against the damages induced by different chemical agents.

Liberação de óxido nítrico e tnf-α em macrófagos peritoneais na presença de Melampodium divaricatum (Asteraceae)

The immune responses are mediated by a variety cells and molecules that cells secreted. Macrophages are the first cells that participate in the immune response, and, when are activated, release more than hundred compounds at the extracelular medium, such as cytokine (TNF-α) and the intermediate compounds of the nitrogen (NO). In this paper the release of nitric oxide (NO) and necrose tumoral factor (TNF-α) were determined in peritoneal macrophage cultures of mice in the presence of the 70% ethanolic extract obtained from the flowers of the Melampodium divaricatum (Asteraceae) in the concentrations of 20, 10 and 5 mg/mL. The 70% ethanolic extracts from flowers of the Melampodium divaricatum presented higher liberation of NO and TNF-α in the concentration of the 20 mg/mL when compared with LPS. We conclude that this extract is a potente stimulator of macrophage, could be immunomodulatory activity.

Avaliação do colesterol plasmático em coelhos com hipercolesterolemia induzida e tratados com extrato etanólico de própolis

The propolis (bee glue) is a product rich in flavonoids, which are known for antioxidant activities, a protective action to the lipoproteins LDL-cholesterol against lipid peroxidation. Because they have antioxidant properties, we investigated the effect of the ethanolic extract propolis on the plasma level of cholesterol in rabbits (Oryctolagus cuniculus) submitted to hypercholesterolaemia. The animals were divided into 4 groups. G 1=received commercial feed and water, G 2=received enriched feed and water, G 3=received enriched feed and ethanol, G 4=received enriched feed and ethanolic extract of propolis. The hypercholesterolaemia was induced with commercial feed enriched with egg yolk. The animals received the ethanolic extract propolis at the concentration of 100 mg/kg daily. Weekly, after fast of 14 hours, the samples of blood were collected from the marginal vein of the ear. The plasma was used for the estimation total cholesterol. From the results obtained, we verified that the ethanolic extract propolis significantly reduced the plasma level cholesterol (109,59 mg/dL, p<0,05), compared to the animals treated with ethanol (331,38 mg/dL), and also to those receiving the commercial feed only, with cholesterol at 269,74 mg/dL.

Quick preparative separation of natural naphthopyranones with antioxidant activity by high-speed counter-current chromatography

The natural naphthopyranones paepalantine (1), paepalantine-9O-β-D-glucopyranoside (2) and paepalantine-9-O-β-D-allopyranosyl-(1→6)-O-β-D-glucopyranoside (3) were separated in a preparative scale from the ethanolic extract of the capitula of Paepalanthus bromelioides by high-speed counter-current chromatography (HSCCC). The solvent system used was composed of water-ethanol-ethyl acetate-hexane (10:4:10:4, v/v/v/v). This technique led to the separation of the three different naphthopyranone glycosides in pure form in approximately 7 hours. Paepalantine showed a good antioxidant activity when assayed by the DPPH radical spectrophotometric assay.

Avaliação bioquímica do extrato etanólico de própolis em ratos

Propolis is a natural product collected by honey bees containing, among other biochemical constituents, a variety of flavonoids. Propolis is a folk medicinal employed for treating various diseases. It is alleged to exhibit a broad spectrum of bioactivities. The aim of this study was to evaluate the effect of ethanolic extract of propolis (EEP) of species Plebeia droryana and Scaptotrigonea bipunctata through biochemical parameters. Rats were divided into 4 groups: (G1) untreated; (G2) ethanol treated; (G3) treated EEP of Plebeia droryana; (G4) treated of Scaptotrigonea bipunctata. The EEP (100 mg/kg b. w., daily) was administered orally to the animals, for 30 days. Treatment with EEP for two species showed reduction (p<0,05) in serum alanine aminotransferase, aspartato aminotransferase and alkaline phosphatase activity, compared to control ethanol values. The administration of EEP lowered significantly the serum levels of cholesterol (G3= 48,83±5,7 mg/dL; G4=56,91±6,5 mg/dL) and triacylglycerol (G3=45,17±4,16 mg/dL; G4=46,74± 3,90 mg/dL). The serum concentration of albumin (G3=4,16±0,6 g/dL; G4= 3,61±0,36 g/dL) increased (p<0,05) after the administration of EEP, however, it did not affect total protein and glucose concentration. The data suggest that EEP of two species caused alterations of the biochemical parameters.

Contribuição ao protocolo de controle de qualidade da própolis e de seus extratos

A quality control protocol for the analysis of propolis and its extracts is proposed in the present work. Propolis of Apis mellifera L. bees collected in beehives of an apiary located in the northwest of the Paraná State was triturated and submitted to the following analysis: particles medium size determination, loss of dry, ashes drift, waxes drift, drift of extractive (in water and in ethanol) and total flavonoids drift determination. Propolis ethanolic extracts (96 °GL) at 10% (w/w) and at 30% (w/w) were prepared and submitted to the determination of the pH, relative density, dry residue, alcoholic drift and total flavonoids drift determination. Propolis was analyzed, through High Performance Liquid Cromatography (HPLC). Comparing the obtained results with other works, it was observed that is possible to establish the intervals of values for parameters in order to evaluate the quality of a propolis sample and its extracts.

Ativação de macrófagos por produtos naturais obtidos da Achillea millefolium L. (Asteraceae)

Plants are a valuable source of natural products for the maintenance of human health. The purpose of this paper was the study of immunologic activity of yarrow (Achillea millefolium L.), a largely used plant in popular medicine that has many different properties such as: antiinflammatory, astringent, antiseptic and antispasmodic. Macrophages stimulation was evaluated by the determination of H2O2, NO and TNF-α in supernatants of peritoneal macrophages cultures of mice in the presence of the yarrow leaves extract. The thin layer chromatography of extract was also analyzed, showing rutin. All concentrations showed a moderate release of H2O2 and the concentrations of 6, 8 and 10mg/mL had a higher release of NO. The TNF-a was produced in all concentrations, but the best result was obtained at 4mg/mL. Analyzing the results, it is suggested that the yarrow ethanolic extract can modulate the macrophages activation.

Atividade anti Staphylococcus aureus de extratos de própolis (EP) de Apis mellifera preparados com diferentes concentrações de etanol

Propolis has been used in folk medicine and possesses a broad spectrum of biological activities, specially antibacterial activity. Studies have demonstrated that the composition of propolis extract may have influence in such activity. The goal of this study was to investigate the antibacterial activity of eleven propolis extracts (PE) against sixty one Staphylococcus aureus strains, isolated from newborn clinical specimens. The PE from Apis mellifera were prepared by using pure water and mixtures of water with ethanol at different concentrations (from 0 to 100%), 25g of propolis in 100 mL of solvent, and three days of maceration followed by filtration. Determination of Minimal Inhibitory Concentration (MIC) by agar dilution method was performed and serial concentrations from each PE were achieved (%v/v) in plates containing Mueller Hinton agar. It was possible to verify that the anti S. aureus activity was directly proportional to ethanol concentration and no significant differences were observed among PE with ethanol concentration from 70 to 100%. The MIC 90% values ranged from 0.4 to 0.6% (v/v) and the 70% ethanolic extract were the most efficient to inhibit bacterial growth (MIC 90%=0.42%, v/v). In conclusion, our results suggest that the EP composition and, consequently, the concentration of ethanol used as solvent may influence the antibacterial activity of propolis from A. mellifera.

Effect of Brazilian propolis on experimental hepatotoxicity in rats

Propolis (bee glue) is one of the major hive products of bees and is rich in flavonoids, which are known for their antioxidant activities. The aim of this study was to evaluate the hepatoprotective effects of the ethanolic extract of propolis (EEP) against experimental carbon tetrachloride (CCl 4)-induced liver toxicity in rats by means of biochemical indices. The animals were divided into 4 groups: GI= received mineral oil; GII= CCl 4(4mL/kg; Lp., single dose) treated; GIII= CCl4 (4mL/kg; i.p., single dose) treatment followed by ethanolic extract of propolis (100mg/kg) for gavage from the species Tetragonisca angustula, daily for 3 days and GIV= CCl4 (4mL/kg; i.p., single dose) treatment followed by ethanolic extract of propolis (100mg/kg) for gavage from the species Nannotrigonea testaceicornes, daily, for 3 days. The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), cholesterol and triacylglycerols were estimated after 3 days. CCl 4 caused a maximum increase (p<0,01) above biochemical parameters. As compared to CCl4 group (GII), the EEP (GIII and GIV) showed reduction in cholesterol, triacylglycerol, ALT, AST and alkaline phosphatase activity in the serum. In conclusion, these data indicate that EEP improved the dyslipidaemia, moreover, significantly attenuated increases in serum ALT and AST activities in rats with liver damage induced by carbon tetrachloride.

In vitro release of propolis from gelatin microparticles prepared by spray-drying technique

The purpose of this study was to investigate the in vitro release of propolis from gelatin microparticles. Gelatin microparticles containing propolis extractive solution (PES) were prepared by spray-drying technique. Microparticles with a mean diameter of 2.50 μm and with regular morphology were obtained. The entrapment efficiency of propolis in the microparticles was over 39%. Spray-drying showed to be a feasible method for the preparation of gelatin microparticles containing propolis. Comparing to PES, the in vitro release of propolis from gelatin microparticles in aqueous medium was slower, considering markers 1 and 2. Thus, it was possible to transform a liquid propolis dosage form into a solid one, improving manipulation, packaging and storage and with modified release in aqueous medium, comparatively to the ethanolic extract of the drug.