Development of a Gas Chromatography Method for the Analysis of Copaiba Oil.

A rapid, simple, precise and economic method for the quantification of main compounds of copaiba resin and essential oils (Copaifera langsdorffii Desf.) by gas chromatography (GC) has been developed and validated. Copaiba essential oil was extracted by hydrodistillation from the copaiba resin. Resin derivatization allowed the identification of diterpenes compounds. A gas chromatography-mass spectroscopy (GC/MS) method was developed to identify compounds composing the copaiba resin and essential oil. Then the GC/MS method was transposed to be used with a flame ionization detector (FID) and validated as a quantitative method. A good correlation between GC/MS and GC/FID was obtained favoring method transposition. The method showed satisfactory sensitivity, specificity, linearity, precision, accuracy, limit of detection and limit of quantitation for ß-caryophyllene, α-humulene and caryophyllene oxide analyses in copaiba resin and essential oils. The main compounds identified in copaiba essential oil were ß-bisabolene (23.6%), ß-caryophyllene (21.7%) and α-bergamotene (20.5%). Copalic acid methyl ester (15.6%), ß-bisabolene (12.3%), ß-caryophyllene (7.9%), α-bergamotene (7.1%) and labd-8(20)-ene-15,18-dioic acid methyl ester (6.7%) were diterpenes identified from the derivatized copaiba resin. The proposed method is suitable for a reliable separation, identification and quantification of compounds present in copaiba resin and essential oil. It could be proposed as an analytical method for the analysis of copaiba oil fraction in raw and essential oil parent extracts and after they have been incorporate in pharmaceutical formulations.

Aqueous Leaf Extract of Jatropha mollissima (Pohl) Bail Decreases Local Effects Induced by Bothropic Venom.

Snakebites are a serious worldwide public health problem. In Brazil, about 90% of accidents are attributed to snakes from the Bothrops genus. The specific treatment consists of antivenom serum therapy, which has some limitations such as inability to neutralize local effects, difficult access in some regions, risk of immunological reactions, and high cost. Thus, the search for alternative therapies to treat snakebites is relevant. Jatropha mollissima (Euphorbiaceae) is a medicinal plant popularly used in folk medicine as an antiophidic remedy. Therefore, this study aims to evaluate the effect of the aqueous leaf extract from J. mollissima on local effects induced by Bothrops venoms. High Performance Liquid Chromatography with Diode Array Detection analysis and Mass Spectrometry analysis of aqueous leaf extract confirmed the presence of the flavonoids isoschaftoside, schaftoside, isoorientin, orientin, vitexin, and isovitexin. This extract, at 50-200 mg/kg doses administered by intraperitoneal route, showed significant inhibitory potential against local effects induced by Bothrops erythromelas and Bothrops jararaca snake venoms. Local skin hemorrhage, local edema, leukocyte migration, and myotoxicity were significantly inhibited by the extract. These results demonstrate that J. mollissima extract possesses inhibitory potential, especially against bothropic venoms, suggesting its potential as an adjuvant in treatment of snakebites.

New drug delivery system for corneal administration of mitomycin-C.

PURPOSE: To develop a new corneal release system to deliver optimum amounts of mitomycin-C (MMC) during the perioperative period of photorefractive keratectomy (PRK). SETTING: Ophthalmos S/A, São Paulo, Brazil. DESIGN: Experimental study. METHODS: An in vitro experimental design was developed for studying a new MMC delivery system at a drug concentration of 0.02%. Whatman sterile filter paper disks with a diameter of 8.0 mm were impregnated with MMC solution. After drying, the disks were placed on agar plates seeded with Staphylococcus epidermidis; this was followed by instillation of a drop of sterile water. After 1 minute, the disks were removed and the plates were incubated for 48 hours at 35°C. The mean volume of the drops delivered from regular eyedrop bottles was determined, and the inhibition zone (in millimeters) was correlated with the amount of MMC loaded onto the disks. RESULTS: Analysis of the inhibition zones produced by MMC indicated that 16 µg was the optimum dose to be incorporated in the disks. The mean volume of a drop delivered from eyedrop bottles was 37.7 µL. One minute after the application of a single drop of a balanced salt solution, the system released an adequate concentration of MMC for PRK. CONCLUSION: A new delivery system for MMC was successfully developed for application during photorefractive keratectomy. FINANCIAL DISCLOSURE: Dr. de Souza Lima Filho is the managing director of Ophtalmos S/A. Drs. de Souza Lima Filho, Irochima Pinheiro, and Oréfice have exclusive rights to intellectual property of this invention secured by a patent application filed with the Instituto Nacional da Propriedade Industrial.

Ipomoea asarifolia neutralizes inflammation induced by Tityus serrulatus scorpion venom.

ETHNOPHARMACOLOGICAL RELEVANCE: Envenoming caused by scorpion sting is a serious public health problem. In Brazil, 13,038 accidents caused by venomous animals have been reported. Of this total, 53% of the cases and 14 deaths were caused by scorpions. Furthermore, Tityus serrulatus (Buthidae) is the most dangerous scorpion due to the high toxicity of its venom. The treatment is the common supportive therapy and the serum therapy, but some people do not have access to both therapies and seek healing through the use of medical plants. AIM OF THE STUDY: This study evaluated the ability of the crude extract and fractions from the leaves of Ipomoea asarifolia in neutralizing the main biological effects caused by Tityus serrulatus envenoming in mice. MATERIALS AND METHODS: BALB/c mice were pretreated (i.v.) with 100 µλ of aqueous extracts and fractions dichloromethane, ethyl acetate, and n-butanol (CH2Cl2, EtOAc, and n-BuOH, respectively) of Ipomoea asarifolia, rutin or saline. Then, the animals received 100 µλ (i.p.) of venom of Tityus serrulatus (0.8 mg/kg). After six hours, the peritoneal lavage was performed with PBS and the number cells were determined using a Neubauer chamber. The supernatants were collected for determination of cytokines, such as IL-6, IL-12, and IL-1ß. RESULTS: The aqueous extract, fractions and rutin, at all doses, significantly reduced cell migration, which was endorsed by the reduction of the levels of certain cytokines. CONCLUSION: This is the first study that demonstrated the potential effect of Ipomoea asarifolia against inflammation caused by Tityus serrulatus venom, suggesting that these extracts and/or their bioactive molecules, especially the flavonoid rutin, have potential use in the therapy of this envenomation.