Caio P. Fernandes

HLB value, an important parameter for the development of essential oil phytopharmaceuticals

Essential oils are used primarily as natural preservatives, flavourants and fragrances in cosmetic products. Several pharmacopeias possess monographs of plants which are good sources of essential oils, such as Brazilian Pharmacopeia, including Illicium verum Hook. f., Schisandraceae and Rosmarinus offi cinalis. Since determination of Hydrophile-Lipophile Balance (HLB) value of essential oils appears as a critical step for development of emulsions and other semi-solid formulations, evaluation of required HLB values for I. verum and R. offi cinalis essential oils is the aim of this study. They were obtained by hydrodistillation and several emulsions were prepared by changing emulsifiers. The couple sorbitan oleate/polysorbate 20 provided best emulsions and was used at different ratios, at a total blend concentration of 5% w/w. The lowest mean droplet diameters for R. offi cinalis and I. verum emulsions were obtained at HLB 16.5 (97.12 nm) and 16.7 (246.6 nm), respectively. Moreover, emulsions with R. offi cinalis were finer and presented some bluish reflection, characteristic of nanoemulsions. The lowest turbidity value for R. offi cinalis emulsion was also obtained at HLB 16.5 (0.33). Thus, the present study describes for the first time HLB values for R. offi cinalis (16.5) and I. verum (16.7) essential oils, contributing to their physicochemical characterization and technology development of phytopharmaceuticals.

Development of a Larvicidal Nanoemulsion with Pterodon emarginatus Vogel Oil.

Pterodon emarginatus Vogel is a Brazilian species that belongs to the family Fabaceae, popularly known as sucupira. Its oil has several biological activities, including potent larvicidal property against Aedes aegypti. This insect is the vector of dengue, a tropical disease that has been considered a critical health problem in developing countries, such as Brazil. Most of dengue control methods involve larvicidal agents suspended or diluted in water and making active lipophilic natural products available is therefore considered a technological challenge. In this context, nanoemulsions appear as viable alternatives to solve this major problem. The present study describes the development of a novel nanoemulsion with larvicidal activity against A. aegypti along with the required Hydrophile Lipophile Balance determination of this oil. It was suggested that the mechanism of action might involve reversible inhibition of acetylcholinesterase and our results also suggest that the P. emarginatus nanoemulsion is not toxic for mammals. Thus, it contributes significantly to alternative integrative practices of dengue control, as well as to develop sucupira based nanoproducts for application in aqueous media.

Development of an insecticidal nanoemulsion with Manilkara subsericea (Sapotaceae) extract

BackgroundPlants have been recognized as a good source of insecticidal agents, since they are able to produce their own defensives to insect attack. Moreover, there is a growing concern worldwide to develop pesticides with low impact to environment and non-target organisms. Hexane-soluble fraction from ethanolic crude extract from fruits of Manilkara subsericea and its triterpenes were considered active against a cotton pest (Dysdercus peruvianus). Several natural products with insecticidal activity have poor water solubility, including triterpenes, and nanotechnology has emerged as a good alternative to solve this main problem. On this context, the aim of the present study was to develop an insecticidal nanoemulsion containing apolar fraction from fruits of Manilkara subsericea.ResultsIt was obtained a formulation constituted by 5% of oil (octyldodecyl myristate), 5% of surfactants (sorbitan monooleate/polysorbate 80), 5% of apolar fraction from M. subsericea and 85% of water. Analysis of mean droplet diameter (155.2 ± 3.8 nm) confirmed this formulation as a nanoemulsion. It was able to induce mortality in D. peruvianus. It was observed no effect against acetylcholinesterase or mortality in mice induced by the formulation, suggesting the safety of this nanoemulsion for non-target organisms.ConclusionsThe present study suggests that the obtained O/A nanoemulsion may be useful to enhance water solubility of poor water soluble natural products with insecticidal activity, including the hexane-soluble fraction from ethanolic crude extract from fruits of Manilkara subsericea.

Triterpene Esters and Biological Activities from Edible Fruits of Manilkara subsericea (Mart.) Dubard, Sapotaceae

Manilkara subsericea (Mart.) Dubard (Sapotaceae) is popularly known in Brazil as “guracica.” Studies with Manilkara spp indicated the presence of triterpenes, saponins, and flavonoids. Several activities have been attributed to Manilkara spp such as antimicrobial, antiparasitic and antitumoral, which indicates the great biological potential of this genus. In all, 87.19% of the hexanic extract from fruits relative composition were evaluated, in which 72.81% were beta- and alpha-amyrin esters, suggesting that they may be chemical markers for M. subsericea. Hexadecanoic acid, hexadecanoic acid ethyl ester, (E)-9-octadecenoic acid ethyl ester, and octadecanoic acid ethyl ester were also identified. Ethanolic crude extracts from leaves, stems, and hexanic extract from fruits exhibited antimicrobial activity against Staphylococcus aureus ATCC25923. These extracts had high IC50 values against Vero cells, demonstrating weak cytotoxicity. This is the first time, to our knowledge, that beta- and alpha-amyrin caproates and caprylates are described for Manilkara subsericea.

Inhibitory Effect of Plant Manilkara subsericea against Biological Activities of Lachesis muta Snake Venom

Snake venom is composed of a mixture of substances that caused in victims a variety of pathophysiological effects. Besides antivenom, literature has described plants able to inhibit injuries and lethal activities induced by snake venoms. This work describes the inhibitory potential of ethanol, hexane, ethyl acetate, or dichloromethane extracts and fractions from stem and leaves of Manilkara subsericea against in vivo (hemorrhagic and edema) and in vitro (clotting, hemolysis, and proteolysis) activities caused by Lachesis muta venom. All the tested activities were totally or at least partially reduced by M. subsericea. However, when L. muta venom was injected into mice 15 min first or after the materials, hemorrhage and edema were not inhibited. Thus, M. subsericea could be used as antivenom in snakebites of L. muta. And, this work also highlights Brazilian flora as a rich source of molecules with antivenom properties.

Laboratory evaluation of the effects of Manilkara subsericea (Mart.) Dubard extracts and triterpenes on the development of Dysdercus peruvianus and Oncopeltus fasciatus

BACKGROUND Studies were carried out to evaluate the effects of Manilkara subsericea extracts and triterpenes on the development of two species of agricultural pest insects, Oncopelus fasciatus and Dysdercus peruvianus. RESULTS All treatments of insects with M. subsericea extracts induced mortality, delayed development and inhibited moulting. Some extracts assayed (FH, FB and FD in D. peruvianus, and FH, FB and FEA in O. fasciatus) also produced body deformities in the few adults that emerged. Other extracts (FH, FEA, FB, FD and LET in both insects), however, induced either permanent (overaged) or extranumerary nymphs, both of which were unable to achieve the adult stage and reproductive status. The insects were also treated with triterpenes (α- and β-amyrin acetates) which showed high lethality at 30 days after treatment and delayed the intermoult period so that overaged nymphs were detected. CONCLUSION The results indicate that extracts and triterpenes from Manilkara subsericea act as potent growth inhibitors of phytophagous hemipteran nymphs. It is also concluded that the mixture of several different molecules in the extracts used produces synergic effects that do not occur after using the triterpenes (PFT) alone. These secondary metabolites in the M. subsericea extracts can potentially be used in integrated control programmes against crop pests.

Essential Oils from Fruits with Different Colors and Leaves of Neomitranthes obscura (DC.) N. Silveira: An Endemic Species from Brazilian Atlantic Forest

Neomitranthes obscura (DC.) N. Silveira is an endemic plant of Brazilian Atlantic Forest and widely spread in the sandbanks of “Restinga de Jurubatiba” National Park. It is popularly known by local population as “camboim-de-cachorro” or “cambuí-preto” and recognized by its black ripe fruits. However, specimens with yellow ripe fruits were localized in the “Restinga de Jurubatiba” National Park. The aim of the present study was to evaluate chemical composition of essential oils obtained from leaves and fruits of N. obscura specimens with different fruit color (black and yellow) by GC and GC-MS. Essential oils from leaves of specimens with black and yellow fruits indicated a predominance of sesquiterpenes (81.1% and 84.8%, resp.). Meanwhile, essential oil from black fruits presented a predominance of monoterpenes (50.5%), while essential oil from yellow fruits had sesquiterpenes (39.9%) as major substances. Despite previous studies about this species, including essential oil extraction, to our knowledge this is the first report on N. obscura fruits with different colors. Our results suggest the occurrence of unless two different varieties for this species.

Baccharis reticularia DC. and Limonene Nanoemulsions: Promising Larvicidal Agents for Aedes aegypti (Diptera: Culicidae) Control

Baccharis reticularia DC. is a plant species from the Asteraceae family that is endemic to Brazil. Despite the great importance of Baccharis genus, no study has been carried out regarding either the phytochemical composition of B. reticularia or the evaluation of its larvicidal potential. Considering the intrinsic immiscibility of essential oils, this study shows larvicidal nanoemulsions containing the B. reticularia phytochemically characterized essential oil and its main constituent against Aedes aegypti. The major compound found was d-limonene (25.7%). The essential oil inhibited the acetylcholinesterase, one of the main targets of insecticides. The required hydrophile-lipophile balance of both nanoemulsions was 15.0. The mean droplet sizes were around 90.0 nm, and no major alterations were observed after 24 h of preparation for both formulations. After 48 h of treatment, the estimated LC50 values were 118.94 μg mL−1 and 81.19 μg mL−1 for B. reticularia essential oil and d-limonene nanoemulsions, respectively. Morphological alterations evidenced by scanning electron micrography were observed on the larvae treated with the d-limonene nanoemulsion. This paper demonstrated a simple and ecofriendly method for obtaining B. reticularia essential oil and d-limonene aqueous nanoemulsions by a non-heating and solvent-free method, as promising alternatives for Aedes aegypti control.

Antioxidant and Hepatoprotective Activity of a New Tablets Formulation from Tamarindus indica L.

Hepatotoxic chemicals damage liver cells primarily by producing reactive oxygen species. The decoction of the leaves of Tamarindus indica L. is used for liver disorders. In this work we evaluated the hepatoprotective activity of a tablet formulation of this plant. Thirty-five Sprague Dawley rats were randomly divided into five groups (n = 7). First group (I) is control group, fed with standard diet. Groups II to V (hepatotoxic groups) were subjected to a subcutaneous injection of CCl4 (0.5 mL/kg). Group II was negative control, fed with standard diet; group III was subjected to administration of Silymarin 150 mg/kg and groups IV and V were treated with tablets in dose of 100 mg/kg and 200 mg/kg, respectively. Lipid peroxidation and the activity of superoxide dismutase, catalase, and reduced glutathione were evaluated. Serum levels of alanine aminotransferase, aspartate aminotransferase, gamma-glutamine transferase, alkaline phosphatase, and a lipid profile were evaluated too. The tablets inhibit lipid peroxidation. The redox balance (SOD-CAT-GSH) remains normal in the experimental groups treated with tablets. The liver function using dose of 200 mg/kg of tablets was better than the other experimental groups. These results justify, scientifically, the ethnobotanical use of the leaves of Tamarindus indica L.

Development and Characterization of Cassia grandis and Bixa orellana Nanoformulations.

Cassia grandis and Bixa orellana are important plant species with folk use and great potential for phytopharmaceuticals. Nanodispersions are disperse systems of insoluble or immiscible substances in a liquid medium that may be prepared with or without coating polymers. To our knowledge, no studies were carried in order to achieve coating-polymer free nanoformulations using B. orellana extract or any C. grandis-based nanoformulations. Thus, on the present study we aimed to develop C. grandis nanoformulations using three different coating polymers (Eudragit® L 100 55, PEG 4000 and Kollicoat®), while B. orellana nanodispersions were obtained using different surfactants (polysorbate 80, polysorbate 20, polyethylene glycol 400 monooleate, polyethylene glycol 600 monooleate, polyethylene glycol 400 dioleate and polyethylene glycol 600 dioleate) as coating polymer-free nanoformulations. Characterization of nanoformulations was performed by different parameters, including particle size, polydispersity index and zeta-potential. Our results suggested that some optimal nanoformulations were obtained for both plant species. Moreover, possible stable behavior was observed during storage period for C. grandis (30 days) and B. orellana (21 days). On this context, the present study contributes to nanobiotechnology development of phytopharmaceuticals, allowing achievement of novel nano-delivery systems with two important folk medicinal plant extracts and making them potential products for innovative phytopharmaceuticals.