Resumo Piper aduncum L., uma planta medicinal brasileira, é conhecida por suas propriedades bioativas, incluindo efeitos repelentes e inseticidas. Este estudo investigou o potencial inseticida dos óleos essenciais (OEs) de P. aduncum, coletados na estação seca e chuvosa, contra pulgas (Ctenocephalides felis felis Bouché, 1835) em estágios de ovos e adultos. Os OEs foram obtidos por hidrodestilação em aparelho de Clevenger, modificado por 2 horas. Análises qualitativas e quantitativas foram realizadas por meio de cromatografia gasosa. Os resultados revelaram que o dilapiol foi a substância predominante, representando 77,56% (chuvoso) e 85,52% (seco) dos OEs. Esses OEs mostraram alta eficácia contra o parasita C. felis felis, resultando em 100% de mortalidade de ovos a uma concentração de 100 μg/mL e 100% de mortalidade para pulgas adultas a partir de 1.000 μg/mL. O padrão dilapiol também foi eficaz, mas em uma concentração relativamente alta. Este achado sugere que os OEs de P. aduncum exibem citotoxicidade contra esses parasitas e podem ter potencial para a produção comercial, oferecendo aplicações práticas para essa bioprospecção. Esse estudo revelou de maneira única que os OEs de P. aduncum, ricos em dilapiol, demonstraram atividade pulicida contra o parasita C. felis felis, inibindo, especialmente, a eclosão dos ovos desses parasitas. L L. brasileira bioativas inseticidas (OEs P chuvosa Ctenocephalides Bouché 1835 adultos Clevenger horas gasosa predominante 7756 77 56 77,56 chuvoso (chuvoso 8552 85 52 85,52 seco (seco C 10 μgmL μg mL 1000 1 000 1.00 eficaz comercial bioprospecção inibindo especialmente 183 775 7 5 77,5 855 8 85,5 00 1.0 18 77, 85, 0 1.

Abstract Piper aduncum L., a Brazilian medicinal plant, is known for its bioactive properties, including repellent and insecticidal effects. This study investigated the insecticidal potential of essential oils (EOs) from P. aduncum, collected during the dry and rainy seasons, against fleas (Ctenocephalides felis felis Bouché, 1835) in egg and adult stages. The EOs were obtained by hydrodistillation using a modified Clevenger apparatus for 2 h. Qualitative and quantitative analysis were performed via gas chromatography. The findings revealed that dillapiole was the predominant substance in both EOs, accounting for 77.6% (rainy) and 85.5% (dry) of the EOs. These EOs exhibited high efficacy against the parasite C. felis felis, resulting in 100% egg mortality at a concentration of 100 μg/mL and 100% mortality for adult fleas starting from 1,000 μg/mL. Dillapiole standard was also effective but at a relatively high concentration. This finding suggested that EOs from P. aduncum exhibit cytotoxicity against these pests and might hold potential for commercial production, offering practical applications for such bioprospecting. This study uniquely revealed that the EOs from P. aduncum, which is rich in dillapiole, demonstrated pulicidal activity against the parasite C. felis felis, particularly in inhibiting the hatching of the eggs of these parasites. L L. plant properties effects (EOs P seasons Ctenocephalides Bouché 1835 stages h chromatography 776 77 6 77.6 (rainy 855 85 5 85.5 (dry C 10 μgmL μg mL 1000 1 000 1,00 production bioprospecting parasites 183 7 77. 8 85. 00 1,0 18 0 1,

This work describes the first determination of the absolute configuration (AC) of (−)-cubebin by means of electronic circular dichroism (ECD), supported by quantum chemical calculations. The comparison of experimental ECD with the corresponding quantum chemical prediction for the proper diastereoisomer resulted in the definitive assignment of the AC of the naturally occurring (−)-cubebin as (8R,8aR,9S). The challenging determination of the relative configuration (RC) of cubebin based only on experimental nuclear magnetic resonance (NMR) methods is stressed. Computation of the 13C and 1H NMR chemical shifts for all the possible diastereoisomers leads to values of mean absolute error and root mean square deviation that do not allow distinguishing among them. Thus, errors in stereochemical determination can easily occur even when using two-dimensional methods, which clearly demonstrate the complexity of this special case. To determine the RC of this bioactive natural compound with high level of confidence, it was necessary to combine the DP4+ method with X-ray crystallography. Therefore, employing the commonly used empirical methods to determine the AC of (−)-cubebin can easily lead to misassignment of its stereochemistry.

Abstract In this study, we developed and validated a chromatographic method by High-performance Liquid Chromatography coupled to Diode-Array Ultraviolet Detector for quantification of acetaminophen in small volumes of plasma. This analytical method is particularly suitable for studies of gastric emptying time and pharmacokinetics in small rodents. Orally administered acetaminophen is promptly and completely absorbed in the intestines, with negligible absorption from the stomach. Owing to these kinetic features, acetaminophen can be used to study gastric emptying. The newly-validated analytical method was employed to investigate whether plants used as anti-dyspeptics in the Brazilian traditional medicine affect the gastric emptying. Lyophilized aqueous extracts from leaves of Baccharis trimera (Less) DC., Asteraceae (carqueja), Maytenus ilicifolia Mart. Ex Reissek, Celastraceae (espinheira-santa) and Lippia alba (Mill.) N.E.Br. ex Britton & P. Wilson, Verbenaceae (erva-cidreira) were administered by gavage to female Wistar rats 30 min before a single oral dose of acetaminophen (50 mg/kg). L. alba extract (50 mg/kg) did not alter acetaminophen plasma concentration versus time curve an indication that it has no effect on gastric emptying. Extracts of B. trimera (50 mg/kg) and M. ilicifolia (30 and 50 mg/kg, but not 10 mg/kg), however, slightly delayed gastric emptying. M. ilicifolia given by intraperitoneal route (30 mg/kg) also retarded gastric emptying. In conclusion, the newly-validated analytical method is adequate for studying the effects of medicinal plants on gastric emptying.

ABSTRACT Casearia genus (Salicaceae) is found in sub-tropical and tropical regions of the world and comprises about 160-200 species. It is a medicinal plant used in South America, also known as “guaçatonga”, “erva-de-tiú”, “cafezinho-do-mato”. In Brazil, there are about 48 species and 12 are registered in the State of Rio de Janeiro, including Casearia sylvestris Sw. There are many studies related to the chemical profile and cytotoxic activities of extracts from these plants, although few studies about the antifungal potential of the essential oil have been reported. In this work, we have studied the antifungal properties of the essential oil of C. sylvestris leaves, as well as of their fractions, against four yeasts (Saccharomyces cerevisae, Candida albicans, C. glabrata and C. krusei) for the first time. The chemical analysis of the essential oil revealed a very diversified (n = 21 compounds) volatile fraction composed mainly of non-oxygenated sesquiterpenes (72.1%). These sesquiterpenes included α-humulene (17.8%) and α-copaene (8.5%) and the oxygenated sesquiterpene spathulenol (11.8%) were also identified. Monoterpenes were not identified. The fractions are mainly composed of oxygenated sesquiterpenes, and the most active fraction is rich in the sesquiterpene 14-hydroxy -9-epi-β-caryophyllene. This fraction was the most effective in inhibiting the growth of three yeast strains.

ABSTRACT Piperaceae species can be found worldwide in tropical and subtropical areas and many of them have been used for centuries in traditional folk medicine and in culinary. In Brazil, species of Piperaceae are commonly used in some communities as local anesthetic and analgesic. Countrified communities have known some species of the genus Ottonia as "anestesia" and it is a common habit of chewing leaves and roots of Ottonia species to relief toothache. The purpose of this study is to report our findings on new molecules entities obtained from the roots of Ottonia anisum Spreng, in which local anesthetic activity (sensory blockage) is demonstrated for the first time in vivo guinea pig model. Phytochemical investigation led to the isolation of three amides (pipercallosidine, piperine and valeramide) and in an enriched mixture of seven amides (valeramide, 4,5-dihydropiperlonguminine, N-isobutil-6-piperonil-2-hexenamide, piperovatine, dihydropipercallosidine, pipercallosidine and pipercallpsine). Our findings demonstrated the anesthetic potential for the methanolic extract from roots, its n-hexane partition and amides from O. anisum and it is in agreement with ethnobotanical survey.

In the European Union, traditional herbal medicines that are regarded as "acceptably safe, albeit not having a recognized level of efficacy" fit into a special category of drugs ("traditional herbal medicine products") for which requirements of non-clinical and clinical studies are less rigorous. A regulation proposal published by the Brazilian National Health Surveillance (Anvisa) defines a similar drug category ("traditional phytotherapeutic products") for registration purposes. Regarding herbal medicines, both agencies seem to be lenient regarding proof of efficacy, and consider long-standing folk use as evidence of safety and a waiver of a thorough toxicological evaluation. Nonetheless, several herbal products and constituents with a long history of folk usage are suspected carcinogenic and/or hepatotoxic. Herbal products have also been shown to inhibit and/or induce drug-metabolizing enzymes. Since herbal medicines are often used in conjunction with conventional drugs, kinetic and clinical interactions are a cause for concern. A demonstration of the safety of herbal medicines for registration purposes should include at least in vitroand in vivogenotoxicity assays, long-term rodent carcinogenicity tests (for drugs intended to be continuously used for > 3 months or intermittently for > 6 months), reproductive and developmental toxicity studies (for drugs used by women of childbearing age), and investigation of the effects on drug-metabolizing enzymes.

Piper aduncum L., Piperaceae, stands out due to its biological activities, however, it is still found in the wild and little is known about its agronomic point of view. The aim of this study was to evaluate the growth and to analyze the chemical composition of essential oils from leaves of P. aduncum collected in two different sites of Cerrado as well as in cultivated plants. The cultivation was installed out in a greenhouse using cuttings of adult specimens. Essential oils were obtained from fresh leaves. Plants from the two studied locations showed erect growth habit and behavior of linear growth. The essential oils composition of P. aduncum from Bocaiuva did not differ between wild and cultivated plants, as the major substance identified as 1,8-cineole. The plants from Montes Claros site showed a distinct concentration for the two samples, being the major substance characterized as transocimene (13.4%) for wild and 1,8-cineole (31.3%) for cultivated plants. Samples from both locations showed a similar essential oil composition in cultivars. Our results showed that P. aduncum cultivation is feasible and the variation in chemical composition of the two sites may indicate an environmental influence, since chemical and isoenzyme analysis did not show great differences.

RESUMOPiper aduncum L. é utilizada na medicina popular para o tratamento de doenças respiratórias e inflamatórias. O objetivo desse estudo foi analisar o óleo essencial das folhas de P. aduncum coletadas no Cerrado brasileiro, Norte de Minas Gerais, assim como avaliar a atividade larvicida desse óleo e de sua substância majoritária. O óleo essencial foi analisado por cromatografia em fase gasosa acoplada à espectrometria de massas e por cromatografia em fase gasosa acoplada a detector por ionização de chamas, que permitiu a caracterização de 23 substâncias (monoterpenos: 90,4%; sesquiterpenos: 7,0%). O componente majoritário foi o 1,8-cineol (53,9%). O óleo essencial desse exemplar mostrou ser muito diferente do óleo da mesma espécie. Larvas de Aedes aegypti foram expostas a diferentes concentrações do óleo essencial e do 1,8-cineol. Após 24h do tratamento com o óleo registrou-se uma taxa de mortalidade de 100% das larvas nas concentrações de 500 e 1,000 ppm. Após 48h a taxa de mortalidade foi de 80% e 50% para as concentrações de 250 e 100 ppm, respectivamente. A CL50 obtida após 24h foi estimada em 289,9 ppm e após 48h foi de 134,1 ppm. A substância majoritária 1,8-cineol não apresentou atividade larvicida.

ABSTRACTPiper aduncum L. is used in folk medicine to treat respiratory and inflammatory diseases. The aim of this study was to analyze the essential oil from leaves ofP. aduncum collected in the Brazilian Cerrado, North of Minas Gerais, as well as to evaluate the larvicidal activity of this oil and of its major constituent. The essential oil was analyzed by gas chromatography coupled to flame ionization detector and gas chromatography coupled to mass spectrometry that allowed characterizing 23 compounds (monoterpenes: 90.4%; sesquiterpenes: 7.0%). The major component was 1,8-cineole (53.9%). This oil showed to be very different from those obtained from the same species. Larvae of A. aegypti were exposed to different concentrations of the essential oil and 1,8-cineole. The mortality rate of 100% was obtained after 24h of treatment with the oil at concentrations of 500 and 1,000 ppm. After 48h of treatment, the mortality rate was 80% and 50% for concentrations of 250 and 100 ppm, respectively. The LC50 obtained after 24h was estimated in 289.9 ppm and after 48h was 134.1 ppm. The major compound 1,8-cineole showed no larvicidal activity.

Proanthocyanidins from P. contorta leaves and from a commercial quebracho extract were isolated and characterized. Flavonoids, catechins and gallic acid were also identified in the extracts of P. contorta. Compounds were evaluated for their antioxidant properties and for their antiviral activity against an acyclovir-resistant herpes simplex virus type 1 strain. The low molecular weight phenolic derivatives and the proanthocyanidins from P. contorta showed the highest antioxidant activity. Purified proanthocyanidins from both P. contorta and quebracho showed the same maximum non toxic concentrations (25 µg/mL), with 82.2% and 100% of virus inhibition, respectively.

O extrato bruto em acetona/ água das folhas de Pseudopiptadenia contorta foi purificado rendendo frações e substâncias puras. A atividade antioxidante foi avaliada para o extrato e frações, empregando-se o teste químico do DPPH. As frações contendo taninos condensados e substâncias fenólicas de baixo peso molecular mostraram ser as responsáveis pela atividade antioxidante do extrato bruto das folhas de P. contorta.

O óleo essencial das folhas de Piper solmsianum e seu constituinte majoritário (sarisan) foram testados para avaliar suas influências no comportamento de camundongos. O óleo essencial foi obtido por hidrodestilação em aparelho do tipo Clevenger modificado e analisado por CG/ EM. Essa análise mostrou a presença de monoterpenos, sesquiterpenos e arilpropanóides. Sarisan, um análogo da myristicina, foi isolado do óleo para se efetuar os testes farmacológicos. Emulsões do óleo essencial e do sarisan a 5,0% e 10,0% (v/v) foram utilizadas nos testes. Pentobarbital (30 mg/ kg s.c.) ou diazepam (2,5 mg/ kg s.c.) foram testados como padrões na verificação de efeitos depressores e ansiolíticos, respectivamente. O óleo essencial e seu componente majoritário apresentaram efeitos excitantes e depressores nos animais testados.

The essential oil from Piper solmsianum leaves and its major compound (sarisan) were tested to verify their influences upon mice behaviour. The essential oil was obtained by hydrodistillation in a modified Clevenger extractor and analysed by GC/ MS. This analysis revealed in the oil the presence of monoterpenes, sesquiterpenes and of arylpropanoids. The compound sarisan, a myristicin analogue, was isolated from the oil to perform the pharmacological tests. Emulsions of the oil and of sarisan (5.0 and 10.0% v/v) were used in the tests. Pentobarbital (30 mg/ kg s.c.) or diazepam (2.5 mg/ kg s.c.) were tested as standard drugs to verify depressant or anxiolytic effects, respectively. Both essential oil and sarisan showed to have exciting and depressant effects in the tested animals.