Maurício Passos

Profiling Propolis Flavonoids by Means of Micellar Electrokinetic Capillary Chromatography, Capillary Gas Chromatography and Bactericidal Action

SummaryThere is no proportional correlation between increasing organic solvent polarity from hexane to methanol and the extractability of propolis solids by the solvents or the bactericidal action of the propolis extracts obtained. Hexane (17.7%) and chloroform (62.1%) were the poorest and the best extractants, respectively, for propolis solids. The antibiotic activity of the extracts againstStaphylococcus strains decreased in the order hexane≥ethanol>methanol. Different capillary GC profiles were obtained for persilyl derivatives of propolis extracted with hexane and methanol and for propolis collected in different Brazilian provinces, suggesting the influence of flora variability on propolis composition. Wavelength-selected DAD detection revealed MEKC to be an innovative and sensitive method for monitoring the occurrence in propolis of the flavonoids and phenolcarboxylic acids thought to be responsible for the antimicrobial activity of propolis.

Fructo-oligosaccharide production from inulin through partial citric or phosphoric acid hydrolyses.

Purified inulin from Dahlia tubers was partially hydrolyzed to form fructo-oligosaccharides by using citric or phosphoric acids (pH, 2.0-2.5) as mild acid catalysts. The ideal kinetic conditions to ensure a high yield of fructo-oligosaccharides relative to free fructose were a temperature range of 85°C-95°C, a hydrolysis time of 15-25 minutes, and a catalyst pH of 2.5. At the higher temperature and the longest hydrolysis time, an inversion of the product ratio occurred. Under these conditions, co-generation of hydroxymethylfurfural occurred, and it was eliminated by activated charcoal. Unlike in classic hydrolysis with hydrochloric or sulfuric acid, deionization of the actual hydrolysates was not necessary because the catalyst neutralization with common bases results in the formation of co-nutrients with alternative uses as foods or fermentation substrates. These whole hydrolysates can be advantageously added as nutraceuticals to carbonated beverages and acidic foods, such as soft drinks and yogurts.

Selective Polarity- and Adsorption-Guided Extraction/Purification of Annona sp. Polar Acetogenins and Biological Assay Against Agricultural Pests

Annonaceae acetogenins (AG) comprise a family of natural chemical modifications of long-chain fatty acids (C35-37) bearing one to several hydroxyls (less often oxo), middle-chain tetrahydrofuran rings, and a 7-lactonized, α/β-unsaturated carboxyl group. Acetogenins’ strong biological activity as larvicides, pesticides, and antitumorals is dependent on these structural variations. The hydroxylation degree is particularly important for these etfects. Seeds, albeit rich in fats (mostly triacylglycerols, [TAG]), are a nonpredatory source of these drugs as compared to other botanical parts such as roots and stems. Conventional lipid extractions lead to quantitative lipid recovery and then the unfavorable natural ratio of TAG:AG in the range >90:<0.1 These extracts thus require, for instance, partitions and extensive silica gel column Chromatographic steps, in order to enrich or purify the AG fraction(s). Great operational difficulties result from the similar polarity and mol. wt. range of TAG and AG when carrying out these purification steps. An alternative fast two-step procedure to obtain polar acetogenin (pAG)-enriched preparations was developed. The extraction procedure for Annona spp. seeds pAG was carried out with acetonitrile (E° = 0.65; log P = - 0.33) as a polar organosolvent, followed by the adsorption of the solvent-free extract on activated charcoal, then washed with hexane and/or chloroform (E° = 0.0 and 0.40: log P = 3.5 and 2.0) for most of the contaminating TAG removal, and then with acetone (E° = 0.56; log P = - 0.23) to the desorption of an enriched pAG fraction. An alternative procedure for pAG extraction was supercritical fluid extraction (SFE) at moderate thermopressurization conditions (65-82°C; 120-130 atm) using CO2 with 10% acetonitrile as the polarity modifier. The pAG fractions’ bioactivity was evaluated with the brine-shrimp test (BST), and for feed deterrance, growth inhibition, and lethality against the high-impact agricultural pests Anticarsia gemmatalis and Pseudaletia sequax caterpillars feeding on soya or grass leaves sprayed with a 10% alcohol-stabilized emulsion of pAG.

Synergistic larvicidal effect and morphological alterations induced by ethanolic extracts of Annona muricata and Piper nigrum against the dengue fever vector Aedes aegypti

BACKGROUND Phytopesticide combinations of different botanical sources are seldom reported. Annona muricata seed and Piper nigrum fruit ethanolic extracts enriched in acetogenins and piperamides, respectively, were synergistically used as larvicides against the dengue fever vector Aedes aegypti. RESULTS Individual bioassays of A. muricata and P. nigrum indicated respective LC50 values of 93.48 and 1.84 µg mL(-1) against third-instar larvae. Five combinations of different proportions of plant extracts pointed to synergism between the extracts. The best A. muricata:P. nigrum extract combination was 90:10, which showed 5.12 times the amount of synergism, as confirmed by statistical equations and total concentration log versus combination proportions. Concerning the morphology, A. muricata caused larvae body elongation, mainly in the abdomen, along with the appearance of a cervix. Conversely, P. nigrum induced abdomen and whole body shortening. The morphological effects of A. muricata were prevalent in all of the combinations tested, irrespective of its proportion in the combination. CONCLUSION It is suggested that the different mechanisms of action of the larvicidal actives A. muricata acetogenins and P. nigrum piperamides explain the observed synergism. The combination of inexpensive botanicals and a low-cost organosolvent such as ethanol leads to a simple and efficient phytolarvicidal formulation.

The combined action of phytolarvicides for the control of dengue fever vector, Aedes aegypti

Amongst other botanical sources, Annona muricata L., Annonaceae, seeds and Piper nigrum L., Piperaceae, fruits are particularly enriched with acetogenins and piperine-related amides, respectively. These crude ethanolic extracts are potent Aedes aegypti bioactives that can kill Aedes aegypti larvae (dengue fever mosquito). A. muricata displayed a 93.48 µg/mL LC50 and P. nigrum an 1.84 µg/mL LC50. An uncommon pharmacognostical/toxicological approach was used, namely different combinations of both extracts to achieve an improved lethal effect on the larvae. The independence test (χ2) was utilized to evaluate the combination of the two crude extracts. All of the tested combinations behaved synergistically and these novel results were attributed to the completely different biochemical mechanisms of the differentiated chemical substances that were present in the two botanical sources. Besides the two above selected plants, Melia azedarach L., Meliaceae, Origanum vulgare L., Lamiaceae, and Ilex paraguariensis A. St.-Hil., Aquifoliaceae, in order of decreasing toxicity, may also be sought as potential extracts for the sake of synergic combinations.

Cassava Starch Maltodextrinization/Monomerization through Thermopressurized Aqueous Phosphoric Acid Hydrolysis

Kinetic conditions were established for the depolymerization of cassava starch for the production of maltodextrins and glucose syrups. Thin-layer chromatography and high-performance liquid chromatography analyses corroborated that the proper H3PO4 strength and thermopressurization range (e.g., 142-170 degrees C; 2.8-6.8 atm) can be successfully explored for such hydrolytic purposes of native starch granules. Because phosphoric acid can be advantageously maintained in the hydrolysate and generates, after controlled neutralization with ammonia, the strategic nutrient triplet for industrial fermentations (C, P, N), this pretreatment strategy can be easily recognized as a recommended technology for hydrolysis and upgrading of starch and other plant polysaccharides. Compared to the classic catalysts, the mandatory desalting step (chloride removal by expensive anion-exchange resin or sulfate precipitation as the calcium-insoluble salt) can be avoided. Furthermore, properly diluted phosphoric acid is well known as an allowable additive in several popular soft drinks such as colas since its acidic feeling in the mouth is compatible and synergistic with both natural and artificial sweeteners. Glycosyrups from phosphorolyzed cassava starch have also been upgraded to high-value single-cell protein such as the pigmented yeast biomass of Xanthophyllomyces dendrorhous (Phaffia rhodozyma), whose astaxanthin (diketo-dihydroxy-beta-carotene) content may reach 0.5-1.0 mg/g of dry yeast cell. This can be used as an ideal complement for animal feeding as well as a natural staining for both fish farming (meat) and poultry (eggs).