Antonio Salatino

Analysis of propolis: some parameters and procedures for chemical quality control

SUMMARYA set of parameters and respective procedures for the establishment of chemical profiles of samples of tinctures and crude propolis is presented. It is proposed that estimations of the content of total phenolic substances, flavonoids, waxes, ash, volatile substances and dry residue be used as parameters to characterize samples of crude propolis. For tinctures, the estimations of total phenolic substances, flavonoids, waxes, specific gravity and ethanol are proposed. Total phenolic substances and flavonoids are measured by spectrophotometric methods, waxes gravimetrically and ethanol by gas chromatography. The accuracy of the spectrophotometric procedures was tested by assaying a mixture with a known composition of phenolic acids and flavonoids. The use of the procedures is exemplified by the analyses of six samples of crude propolis from different localities in Brazil and of tinctures prepared with absolute and 70% aqueous ethanol. The contents of total phenolic substances, flavonoids, waxes and vo...

Traditional uses, chemistry and pharmacology of Croton species (Euphorbiaceae)

Croton is a genus of Euphorbiaceae comprising around 1,300 species, widespread in tropical regions of the Old and New Worlds. Several species have a long role in the traditional use of medicinal plants in Africa, Asia and South America. Popular uses include treatment of cancer, constipation, diabetes, digestive problems, dysentery, external wounds, fever, hypercholesterolemia, hypertension, inflammation, intestinal worms, malaria, pain, ulcers and weight-loss. Several species of Croton have a red sap, in some species containing proanthocyanidins and/or alkaloids. The latter may be taspine or some of several benzylisoquinoline-like compounds. Diterpenes are very common in Croton, corresponding to clerodanes, cembranoid, halimanes, kauranes, labdanes, phorbol esters, trachylobanes and sarcopetalanes. Some species are aromatic due to the possession of volatile oils. Representatives of new classes of compounds (phenylbutanoids, glutarimide alkaloids, sarcopetalane diterpenes) have been isolated from Croton species. While laticifers have been described in Croton species, so far there are no anatomical studies about secretory structures of volatile oil. Few studies about flavonoids have been carried out with Croton species. Chemical affinities are apparent in the genus, grouping species with (i) kauranes and/or labdanes, (ii) trachylobanes and (iii) alkaloids. Pharmacological assays have frequently corroborated the traditional uses of Croton species. A great part of pharmacological assays with Croton substances dealt with the clerodane trans-dehydrocrotonin, a wide diversity of effects having been noticed, including hypolipidemic, hypoglycaemic, anti-oestrogen and anti-cancer. Cytotoxic effects also have been observed in assays with alkaloids (taspine) and with secokaurene, labdane and cembranoid diterpenes. Several other effects of Croton substances have been registered, including anti-hypertensive, anti-inflammatory, antimalarial, antimicrobial, antispasmodic, antiulcer, antiviral and myorelaxant.

Origin and Chemical Variation of Brazilian Propolis.

Propolis is a hive product containing chiefly beeswax and plant-derived substances such as resin and volatile compounds. Propolis has been used as an antiseptic and wound healer since ancient times and interest for the product has increased recently. Probably few plant species contribute as major resin sources. Green propolis derives mainly from vegetative apices of Baccharis dracunculifolia (alecrim plants). However, wide variation detected in the chemical composition suggests contributions from alternative resin plant sources. Predominant components of the resin of green propolis are cinnamic acids, chiefly compounds bearing prenyl groups. Terpenoid compounds, such as sesqui, di and pentacyclic triterpenoids, have been detected in many, but not all, samples investigated. Propolis research has uncovered potentialities of substances previously isolated from plants and has detected constituents of plant origin that would hardly be known otherwise.

Seasonal Variation, Chemical Composition and Antioxidant Activity of Brazilian Propolis Samples

Total phenolic contents, antioxidant activity and chemical composition of propolis samples from three localities of Minas Gerais state (southeast Brazil) were determined. Total phenolic contents were determined by the Folin–Ciocalteau method, antioxidant activity was evaluated by DPPH, using BHT as reference, and chemical composition was analyzed by GC/MS. Propolis from Itapecerica and Paula Cândido municipalities were found to have high phenolic contents and pronounced antioxidant activity. From these extracts, 40 substances were identified, among them were simple phenylpropanoids, prenylated phenylpropanoids, sesqui- and diterpenoids. Quantitatively, the main constituent of both samples was allyl-3-prenylcinnamic acid. A sample from Virginópolis municipality had no detectable phenolic substances and contained mainly triterpenoids, the main constituents being α- and β-amyrins. Methanolic extracts from Itapecerica and Paula Cândido exhibited pronounced scavenging activity towards DPPH, indistinguishable from BHT activity. However, extracts from Virginópolis sample exhibited no antioxidant activity. Total phenolic substances, GC/MS analyses and antioxidant activity of samples from Itapecerica collected monthly over a period of 1 year revealed considerable variation. No correlation was observed between antioxidant activity and either total phenolic contents or contents of artepillin C and other phenolic substances, as assayed by CG/MS analysis.

Plant Origin of Green Propolis: Bee Behavior, Plant Anatomy and Chemistry

Propolis, a honeybee product, has gained popularity as a food and alternative medicine. Its constituents have been shown to exert pharmacological effects, such as anti-microbial, anti-inflammatory and anticancer. Shoot apices of Baccharis dracunculifolia (alecrim plant, Asteraceae) have been pointed out as sources of resin for green propolis. The present work aimed (i) to observe the collecting behavior of bees, (ii) to test the efficacy of histological analysis in studies of propolis botanical origin and (iii) to compare the chemistries of alecrim apices, resin masses and green propolis. Bee behavior was observed, and resin and propolis were microscopically analyzed by inclusion in methacrylate. Ethanol extracts of shoot apices, resin and propolis were analyzed by gas chromatography/mass spectroscopy. Bees cut small fragments from alecrim apices, manipulate and place the resulting mass in the corbiculae. Fragments were detected in propolis and identified as alecrim vestiges by detection of alecrim structures. Prenylated and non-prenylated phenylpropanoids, terpenoids and compounds from other classes were identified. Compounds so far unreported for propolis were identified, including anthracene derivatives. Some compounds were found in propolis and resin mass, but not in shoot apices. Differences were detected between male and female apices and, among apices, resin and propolis. Alecrim apices are resin sources for green propolis. Chemical composition of alecrim apices seems to vary independently of season and phenology. Probably, green propolis composition is more complex and unpredictable than previously assumed.

Epicuticular waxes from caatinga and cerrado species and their efficiency against water loss

The effects of the contents and chemical composition of the foliar epicuticular waxes of species from the caatinga (Aspidosperma pyrifolium, Capparis yco, Maytenus rigida and Ziziphus joazeiro) and cerrado (Aristolochia esperanzae, Didymopanax vinosum, Strychnos pseudoquina and Tocoyena formosa) were evaluated as to the resistance to water loss by means of an experimental device constructed for this purpose. In general, the waxes of the caatinga species investigated were more efficient against water loss than cerrado species. Increase of the thickness of the waxy deposits from 40 to 90 microg.cm-2 had no significant effect on the resistance to water loss. The chemistry of the wax constituents was shown to be an important factor to determine the degree of resistance to evaporation. n-Alkanes and alcoholic triterpenes were the most efficient barriers, while hentriacontan-16-one (a ketone) and ursolic acid (an acid triterpene) revealed low efficiency. The higher efficiency of the waxes of the leaves from caatinga species (mainly those of C. yco and Z. joazeiro) is probably accounted for the predominance of n-alkanes in their composition. The lower efficiency of the waxes of A. pyrifolium (caatinga), T. formosa and A. esperanzae (both species from the cerrado) is probably a consequence of the predominance of triterpenoids in the waxes of the two former species and hentriacontan-16-one in the latter.

Aqueous Extract of Brazilian Green Propolis: Primary Components, Evaluation of Inflammation and Wound Healing by Using Subcutaneous Implanted Sponges

Propolis is a chemically complex resinous bee product which has gained worldwide popularity as a means to improve health condition and prevent diseases. The main constituents of an aqueous extract of a sample of green propolis from Southeast Brazil were shown by high performance liquid chromatography/mass spectroscopy/mass spectroscopy to be mono- and di-O-caffeoylquinic acids; phenylpropanoids known as important constituents of alcohol extracts of green propolis, such as artepillin C and drupanin were also detected in low amounts in the aqueous extract. The anti-inflammatory activity of this extract was evaluated by determination of wound healing parameters. Female Swiss mice were implanted subcutaneously with polyesther-polyurethane sponge discs to induce wound healing responses, and administered orally with green propolis (500 mg kg−1). At 4, 7 and 14 days post-implantation, the fibrovascular stroma and deposition of extracellular matrix were evaluated by histopathologic and morphometric analyses. In the propolis-treated group at Days 4 and 7 the inflammatory process in the sponge was reduced in comparison with control. A progressive increase in cell influx and collagen deposition was observed in control and propolis-treated groups during the whole period. However, these effects were attenuated in the propolis-treated group at Days 4 and 7, indicating that key factors of the wound healing process are modulated by propolis constituents.

Quinolizidine alkaloids as systematic markers of the papilionoideae

Abstract The structural characteristics of quinolizidine alkaloids have been quantified on the basis of their biogenetic origins. Such data have been used to determine evolutionary advancement parameters for genera of the Papilionoideae. These analyses indicate a correlation of the structural characters of alkaloids with ecogeography and morphology of the genera, suggesting an adaptive radiation of the taxa from tropical Africa in three major directions along a southern temperate “pre-cytisine” route, a northern temperatre “pre-cytisine-cytisine” route and a tropical “ormosanine” route with a temperate cytisine branch. The data also suggest the division of the Sophoreae into three chemical groups giving rise to Liparieae, Genisteae, Thermopsideae and Bossiaeeae. These concepts are embodied in a tentative phyletic classification of all genera of the Papilionoideae which contain quinolizidine alkaloids.

Comparative Chemistry of Propolis from Eight Brazilian Localities

Propolis is a complex honeybee product with resinous aspect, containing plant exudates and beeswax. Their color, texture, and chemical composition vary, depending on the location of the hives and local flora. The most studied Brazilian propolis is the green (alecrim-do-campo) type, which contains mainly prenylated phenylpropanoids and caffeoylquinic acids. Other types of propolis are produced in Brazil, some with red color, others brown, grey, or black. The aim of the present work was to determine the chemical profiles of alcohol and chloroform extracts of eight samples of propolis, corresponding to six Brazilian regions. Methanol and chloroform extracts were obtained and analyzed by HPLC/DAD/ESI/MS and GC/MS. Two chemical profiles were recognized among the samples analyzed: (1) black Brazilian propolis, characterized chiefly by flavanones and glycosyl flavones, stemming from Picos (Piauí state) and Pirenópolis (Goiás state); (2) green Brazilian propolis, characterized by prenylated phenylpropanoids and caffeoylquinic acids, stemming from Cabo Verde (Bahia state), Lavras and Mira Bela (Minas Gerais state), Pariquera-Açu and Bauru (São Paulo state), and Ponta Grossa (Paraná state). The present work represents the first report of prenylated flavonoids in Brazilian propolis and schaftoside (apigenin-8-C-glucosyl-6-C-arabinose) in green propolis.

Effects of Auxin, Cytokinin and Ethylene Treatments on the Endogenous Ethylene and Auxin-to-cytokinins Ratio Related to direct Root Tip Conversion of Catasetum fimbriatum Lindl. (Orchidaceae) into Buds

Summary The present investigation attempts to correlate the effect of the exogenous auxins, cytokinins and ethylene with the endogenous ethylene and auxin-to-cytokinin ratio, and with the direct root-to-bud conversion in Catasetum fimbriatum . Incubation of root tips with IAA and NAA strongly inhibited both root elongation and bud formation. On the other hand, treatment with IBA resulted in a conspicuous stimulation of longitudinal root growth, while bud formation was retarded. High levels of exogenous zeatin, iP and BA enhanced bud formation, and inhibited root growth. Ethylene production was measured in IAA, IBA and zeatin-treated roots. All growth regulators elicited ethylene evolution, which decreased considerably along the period of incubation. Addition of AVG to the media reduced ethylene evolution. Endogenous auxin, cytokinins, and ABA contents were determined by means of HPLC and ELISA methods in root tips of C. fimbriatum pre-incubated with ethylene (CEPA) and cytokinin (iP). Both exogenous substances stimulated bud formation and conduced to an endogenous IAA/Cks ratio favourable to Cks. From these results it is proposed that the stimulatory effect of exogenous Cks and ethylene on root-to-bud conversion of C. fimbriatum seems to be mediated by the establishment of an endogenous auxin-to-cytokinins balance favourable to shoot formation.