Development, validation and application of a gas chromatography method for the determination of dillapiole from Piper aduncum essential oil in skin permeation samples.

The essential oil extracted from the leaves of Piper aduncum has antifungal, insecticidal and antibacterial activity. Studies with its main compound, dillapiole (DIL) revealed antibacterial and anti-inflammatory potential. Despite all this bioactivity, there is no updated report on the development and validation of analytical and bioanalytical methodology to quantify DIL in skin samples. A selective, precise, accurate and adequate method for the determination of DIL in solutions, porcine ear skin samples and receptor fluid was developed and validated by headspace extraction-gas chromatography with flame ionization detection (HS-GC-FID). HS-GC-FID was applied to determine DIL in Franz cell permeation and retention studies using porcine ear skin samples. In the HS-GC-FID method, matrix-related interferences were not observed at the peak of the DIL retention time. The results showed a high recovery (>97%) after the extraction procedure, allowing the quantification of DIL in complex matrices. In vitro permeation/retention for DIL showed cumulative amounts permeated in the order: receptor fluid (21.98 ± 1.19 µg/cm2 ) > epidermis (15.40 ± 1.20 µg/cm2 ) > dermis (9.52 ± 1.13 µg/cm2 ). HS-GC-FID was successfully validated and the results point to DIL transdermal permeation and to the potential to develop pharmaceutical formulations for skin delivery to treat inflammation or infections.

Piper aduncum Essential Oil Rich in Dillapiole: Development of Hydrogel-Thickened Nanoemulsion and Nanostructured Lipid Carrier Intended for Skin Delivery.

The essential oil extracted from the leaves of Piper aduncum, an aromatic plant from the Amazon region, is rich in dillapiole and presents anti-inflammatory activity. In this study, nanoemulsions (NE) and nanostructured lipid carriers (NLC), which are biocompatible nanostructured systems of a lipid nature, were prepared by high-pressure homogenization for the yet unexplored skin delivery of dillapiole. The addition of hydroxyethylcellulose produced hydrogel-thickened NE or NLC in view to improving the viscosity and skin adherence of the nanoformulations. Formulations were characterized with respect to dillapiole content, droplet size, polydispersity index, zeta potential, morphology, rheological behavior, bioadhesion, skin permeation profile, and in vitro irritancy (HET-CAM). The formulations developed presented spherical, homogeneous nanometric particle size (around 130 nm), narrow polydispersity index (<0.3), and negative zeta potential (around −40 mV). Dillapiole content was slightly lower in NLC compared to NE since the production process involves heating. The hydrogels containing nanocarriers showed pseudoplastic behavior with bioadhesive characteristics. The developed formulations exhibited a controlled release profile, dillapiole delivery up to the dermis, the layer of interest for anti-inflammatory potential, and low irritant potential in the chorioallantoic membrane (HET-CAM). Both hydrogels-thickened NE and NLC seemed to be promising formulations for skin delivery of Piper aduncum essential oil.

Dillapiole in Piper holtonii as an Inhibitor of the Symbiotic Fungus Leucoagaricus gongylophorus of Leaf-Cutting Ants.

Plants of the Piperaceae family are studied for their diverse secondary metabolism with a vast array of compounds that act as chemical defense agents against herbivores. Of all the agricultural pests, the management of insects is a highly significant challenge in the Neotropics, and ants of the Attini tribe pose a major problem. Due to their symbiotic association with the fungus Leucoagaricus gongylophorus (Möller) Singer (Agaricaceae), the species of Atta and Acromyrmex have exhaustive foraging activity which has intensified as deforestation and monoculture farming have increased. The control of leaf-cutting ants is still carried out with synthetic products with negative consequences to the environment and human health. In search for natural and sustainable alternatives to synthetic pesticides, Piper holtonii C. DC. was selected among other plant species after field observations of the foraging activity of Atta cephalotes, which revealed that P. holtonii was never chosen by ants. In vitro evaluation of an ethanol extract of the leaves of P. holtonii resulted in promising inhibitory activity (IC50 102 ppm) against L. gongylophorus. Subsequently, bioassay-guided fractionation led to the isolation of the phenylpropanoid dillapiole, which was also detected in the essential oil. This compound demonstrated inhibition of the fungus with an IC50 of 38 ppm. Considering the symbiotic relationship between the Attini ants and L. gongylophorus, the negative effect on the survival of one of the organisms will affect the survival of the other, so dillapiole or standardized essential oil extracts of P. holtonii containing this active principle could be a unique and useful source as a control agent for leaf cutting-ants.

Genotoxic Effects of Semi-Synthetic Isodillapiole on Oviposition in Aedes aegypti (Linnaeus, 1762) (Diptera: Culicidae)

Abstract INTRODUCTION: Semi-synthetic dillapiole compounds derived from Piper aduncum essential oil are used as alternative insecticides to control insecticide-resistant Aedes aegypti. Thus, we aimed to evaluate the genotoxic effects of semi-synthetic isodillapiole on the nuclei of neuroblasts (larvae) and oocytes (females) and the mean oviposition rates of the females over four generations (G1, G2, G3, and G4) of Ae. aegypti. METHODS: Larvae were captured in the city of Manaus, Amazonas state, Brazil, and exposed to isodillapiole in bioassays (20, 40, and 60 µg/mL) and a negative control (0.05% DMSO in tap water) for 4 h. The cerebral ganglia were extracted from the larvae and oocytes from the adult females to prepare slides for cytogenetic analysis. Breeding pairs were established and eggs counts were quantified taken after the bioassays. RESULTS: The analysis of 20,000 interphase nuclei of neuroblasts and oocytes indicated significant genotoxicity (micronuclei, budding, polynucleated cells, and other malformations) compared to that of the control. Metaphasic and anaphasic nuclei presented chromosomal breaks; however, no significant variation and damage was observed in the negative control. A significant reduction in mean oviposition rates was also recorded following exposure to isodillapiole over the four generations (G1, G2, G3, and G4). CONCLUSIONS: The toxic and genotoxic effects of isodillapiole on Ae. aegypti were caused by reduced oviposition in the females and nuclear abnormalities over the four generations of the trials. Further studies are required, rather than our in vitro assays, to verify the efficacy of exposure to this compound for controlling Ae. aegypti.

Effects of inoculation by arbuscular mycorrhizal fungi on the composition of the essential oil, plant growth, and lipoxygenase activity of Piper aduncum L.

The aim of this study was to evaluate the changes in the production of secondary metabolites Piper aduncum seedlings were inoculated by spores of the arbuscular mycorrhizal fungi (AMF) Rhizophagus clarus and Claroideoglomus etunicatum. P. aduncum seedlings were inoculated by spores of R. clarus and C. etunicatum and then, development parameters, root colonization, lipoxygenase (LOX) activity, and essential oil (OE) chemical composition were monitored at 30, 60 and 90 days' post-inoculation (dpi). The inoculation had influenced the plant height and root length at 30 and 90 dpi and microscopic analysis of roots showed the presence of hyphae, arbuscules and vesicles in the inoculated plants. Phenylpropanoids and sesquiterpene hydrocarbons were the main compounds in the EO. In the leaves, the concentration of phenylpropanoids showed a decrease, mainly at 60 dpi, with increased sesquiterpene hydrocarbon production. The main compounds were dillapiole, myristicin, and germacrene D; the dillapiole concentration decreased in all treatments. LOX activity had an increase in the leaves and roots at 90 dpi. These results suggest that alterations in the secondary metabolites of P. aduncum can be induced by its mechanisms of resistance during AMF interaction.

First phytochemical description of essential oils from Piper cachimboense (Piperales, Piperaceae) / Primeira descrição fitoquímica de óleos essenciais de Piper cachimboense (Piperales, Piperaceae)

Piper cachimboense is recorded only for the Amazon region of Brazil and Colombia, and the objective of this study was to report the first phytochemical assessment of the composition of the essential oils (EOs) from this species collected in the Amazon rainforest, in Novo Progresso, Pará State, Brazil. Samples of leaves were subjected to hydrodistillation in a Clevenger-type apparatus. The chemical identification was carried out by gas chromatography. The yield of oils was of 11.03 ± 5.94% for fresh leaves, and 1.07 ± 0.27% for dry leaves. The analysis showed 36 volatile compounds from fresh leaves and 49 from dried leaves. Main constituents in EOs of both fresh and dried leaves from P. cachimboense were (E)-caryophyllene, germacrene-D, -amorfene, -cadinene and apiole.(AU)

Piper cachimboense é registrada apenas para a região amazônica do Brasil e Colômbia, e o objetivo deste estudo foi relatar, pela primeira vez, a composição fitoquímica dos óleos essenciais (OEs) desta espécie coletada na floresta amazônica, em Novo Progresso/PA, Brasil. Amostras de folhas foram submetidas a hidrodestilação em aparelho tipo Clevenger. A identificação química foi realizada por cromatografia gasosa e o rendimento dos óleos foi de 11,03 ± 5,94% para as folhas frescas e de 1,07 ± 0,27% para as folhas secas. A análise mostrou 36 compostos voláteis para folhas frescas e 49 para folhas secas. Os constituintes principais dos OEs de folhas frescas e secas de P. cachimboense foram (E)-cariofileno, germacreno-D, -amorfeno, -cadineno e apiol.(AU)

First phytochemical description of essential oils from Piper cachimboense (Piperales, Piperaceae) / Primeira descrição fitoquímica de óleos essenciais de Piper cachimboense (Piperales, Piperaceae)

Piper cachimboense is recorded only for the Amazon region of Brazil and Colombia, and the objective of this study was to report the first phytochemical assessment of the composition of the essential oils (EOs) from this species collected in the Amazon rainforest, in Novo Progresso, Pará State, Brazil. Samples of leaves were subjected to hydrodistillation in a Clevenger-type apparatus. The chemical identification was carried out by gas chromatography. The yield of oils was of 11.03 ± 5.94% for fresh leaves, and 1.07 ± 0.27% for dry leaves. The analysis showed 36 volatile compounds from fresh leaves and 49 from dried leaves. Main constituents in EOs of both fresh and dried leaves from P. cachimboense were (E)-caryophyllene, germacrene-D, -amorfene, -cadinene and apiole.

Piper cachimboense é registrada apenas para a região amazônica do Brasil e Colômbia, e o objetivo deste estudo foi relatar, pela primeira vez, a composição fitoquímica dos óleos essenciais (OEs) desta espécie coletada na floresta amazônica, em Novo Progresso/PA, Brasil. Amostras de folhas foram submetidas a hidrodestilação em aparelho tipo Clevenger. A identificação química foi realizada por cromatografia gasosa e o rendimento dos óleos foi de 11,03 ± 5,94% para as folhas frescas e de 1,07 ± 0,27% para as folhas secas. A análise mostrou 36 compostos voláteis para folhas frescas e 49 para folhas secas. Os constituintes principais dos OEs de folhas frescas e secas de P. cachimboense foram (E)-cariofileno, germacreno-D, -amorfeno, -cadineno e apiol.

Rooting of apical, median and basal stem cuttings of Piper aduncum L. on different substrates / Enraizamento de estacas apicais, medianas e basais de Piper aduncum L. em diferentes substratos

Piper aduncum L. (Piperaceae) is an aromatic species native from Tropical Americas. The elevated dillapiole content in its essential oil attributes great economic potential to the species, due to its proven insecticidal action against important agricultural pests. The domestication and cultivation of this species, seeking the standardization of the essential oil quality, goes through the stage of plant propagation, which presents scarce technical and scientific information. Thus, the objective of this study was to evaluate the rooting of stem cuttings collected from the apical, median and basal positions of P. aduncum plagiotropic branches, in sieved soil, commercial substrate (composed of pine bark, peat, expanded vermiculite and enriched with macro and micronutrients), and vermiculite of medium granulometry. The cuttings were kept under intermittent misting for 45 days until the experimental evaluation. Higher rooting percentages were observed in apical cuttings (17.4%) when compared to basal ones (6.9%). There were also higher number, length and fresh mass of roots in apical cuttings. The substrate did not influence the rooting of P. aduncum stem cuttings.

Piper aduncum L. (Piperaceae), é uma espécie aromática nativa da América Tropical. O alto teor do composto dilapiol no óleo essencial atribui à espécie grande potencial econômico em função de sua comprovada ação inseticida em importantes pragas agrícolas. A domesticação e cultivo da espécie, visando a padronização da qualidade do óleo essencial, passa pela etapa de produção de mudas, na qual são escassas as informações técnicas e científicas. Nesse sentido, objetivou-se com o presente trabalho avaliar o enraizamento de estacas caulinares coletadas das posições apical, mediana e basal de ramos plagiotrópicos de P. aduncum em solo peneirado, substrato comercial (composto de casca de pinus, turfa, vermiculita expandida e enriquecido com macro e micronutrientes) e vermiculita de granulometria média. As estacas foram mantidas sob nebulização intermitente por 45 dias até a avaliação do experimento. Não houve interação significativa entre substratos e tipos de estaca. Avaliando os fatores isoladamente, observaram-se maiores porcentagens de enraizamento em estacas apicais (17,4%) em relação às basais (6,9%). Verificou-se também maiores número, comprimento e massa fresca de raízes nas estacas da porção apical. O substrato não influenciou o enraizamento das estacas de P. aduncum.

Rooting of apical, median and basal stem cuttings of Piper aduncum L. on different substrates / Enraizamento de estacas apicais, medianas e basais de Piper aduncum L. em diferentes substratos

Piper aduncum L. (Piperaceae) is an aromatic species native from Tropical Americas. The elevated dillapiole content in its essential oil attributes great economic potential to the species, due to its proven insecticidal action against important agricultural pests. The domestication and cultivation of this species, seeking the standardization of the essential oil quality, goes through the stage of plant propagation, which presents scarce technical and scientific information. Thus, the objective of this study was to evaluate the rooting of stem cuttings collected from the apical, median and basal positions of P. aduncum plagiotropic branches, in sieved soil, commercial substrate (composed of pine bark, peat, expanded vermiculite and enriched with macro and micronutrients), and vermiculite of medium granulometry. The cuttings were kept under intermittent misting for 45 days until the experimental evaluation. Higher rooting percentages were observed in apical cuttings (17.4%) when compared to basal ones (6.9%). There were also higher number, length and fresh mass of roots in apical cuttings. The substrate did not influence the rooting of P. aduncum stem cuttings.(AU)

Piper aduncum L. (Piperaceae), é uma espécie aromática nativa da América Tropical. O alto teor do composto dilapiol no óleo essencial atribui à espécie grande potencial econômico em função de sua comprovada ação inseticida em importantes pragas agrícolas. A domesticação e cultivo da espécie, visando a padronização da qualidade do óleo essencial, passa pela etapa de produção de mudas, na qual são escassas as informações técnicas e científicas. Nesse sentido, objetivou-se com o presente trabalho avaliar o enraizamento de estacas caulinares coletadas das posições apical, mediana e basal de ramos plagiotrópicos de P. aduncum em solo peneirado, substrato comercial (composto de casca de pinus, turfa, vermiculita expandida e enriquecido com macro e micronutrientes) e vermiculita de granulometria média. As estacas foram mantidas sob nebulização intermitente por 45 dias até a avaliação do experimento. Não houve interação significativa entre substratos e tipos de estaca. Avaliando os fatores isoladamente, observaram-se maiores porcentagens de enraizamento em estacas apicais (17,4%) em relação às basais (6,9%). Verificou-se também maiores número, comprimento e massa fresca de raízes nas estacas da porção apical. O substrato não influenciou o enraizamento das estacas de P. aduncum.(AU)

Piper aduncum against Haemonchus contortus isolates: cross resistance and the research of natural bioactive compounds / Piper aduncum contra isolados de Haemonchus contortus: resistência cruzada e a pesquisa de bioativos naturais

Abstract The anthelminthic activity of the essential oil (EO) of Piper aduncum L. was tested in vitro on eggs and larvae of resistant (Embrapa2010) and susceptible (McMaster) isolates of Haemonchus contortus. The EO was obtained by steam distillation and its components identified by chromatography. EO concentrations of 12.5 to 0.02 mg/mL were used in the egg hatch test (EHT) and concentrations of 3.12 to 0.01 mg/mL in the larval development test (LDT). Inhibition concentrations (IC) were determined by the SAS Probit procedure, and significant differences assessed by ANOVA followed by Tukeys test. In the EHT, the IC50 for the susceptible isolate was 5.72 mg/mL. In the LDT, the IC50 and IC90 were, respectively, 0.10 mg/mL and 0.34 mg/mL for the susceptible isolate, and 0.22 mg/mL and 0.51 mg/mL for the resistant isolate. The EO (dillapiole 76.2%) was highly efficacious on phase L1. Due to the higher ICs obtained for the resistant isolate, it was raised the hypothesis that dillapiole may have a mechanism of action that resembles those of other anthelmintic compounds. We further review and discuss studies, especially those conducted in Brazil, that quantified the major constituents of P. aduncum-derived EO.(AU)

Resumo Este estudo avaliou a atividade anti-helmíntica in vitro do óleo essencial (OE) de Piper aduncum L. sobre ovos e larvas de Haemonchus contortus, verificando se um isolado resistente (Embrapa2010), apresentaria o mesmo comportamento que um sensível (McMaster). O OE foi obtido por arraste a vapor e analisado por cromatografia para identificação dos constituintes. O óleo foi avaliado nas concentrações de 12,5 a 0,02 mg/mL no Teste de eclosão dos ovos (TEO) e nas concentrações de 3,12 a 0,01 mg/mL no Teste de desenvolvimento larvar (TDL). As concentrações inibitórias (CI) foram determinadas pelo procedimento Probit do SAS e as diferenças estatísticas geradas pela ANOVA seguida pelo teste de Tukey. Para o isolado sensível obteve-se CI50 de 5,72 mg/mL no TEO. No TDL o óleo apresentou CI50 e CI90 de 0,10 mg/mL e 0,34 mg/mL para o isolado sensível e 0,22 mg/mL e 0,51 mg/mL para o resistente, respectivamente. Demonstrou-se que o OE (dilapiol 76,2%) teve alta eficácia sobre a fase L1. Devido às elevadas CIs obtidas para o isolado resistente, levantou-se a hipótese de que o dilapiol talvez possua um mecanismo de ação semelhante a algum grupo anti-helmíntico. O artigo faz uma revisão e discute estudos de quantificação dos constituintes majoritários do OE de P. aduncum, destacando os realizados no Brasil.(AU)

Piper aduncum against Haemonchus contortus isolates: cross resistance and the research of natural bioactive compounds / Piper aduncum contra isolados de Haemonchus contortus: resistência cruzada e a pesquisa de bioativos naturais

Abstract The anthelminthic activity of the essential oil (EO) of Piper aduncum L. was tested in vitro on eggs and larvae of resistant (Embrapa2010) and susceptible (McMaster) isolates of Haemonchus contortus. The EO was obtained by steam distillation and its components identified by chromatography. EO concentrations of 12.5 to 0.02 mg/mL were used in the egg hatch test (EHT) and concentrations of 3.12 to 0.01 mg/mL in the larval development test (LDT). Inhibition concentrations (IC) were determined by the SAS Probit procedure, and significant differences assessed by ANOVA followed by Tukey’s test. In the EHT, the IC50 for the susceptible isolate was 5.72 mg/mL. In the LDT, the IC50 and IC90 were, respectively, 0.10 mg/mL and 0.34 mg/mL for the susceptible isolate, and 0.22 mg/mL and 0.51 mg/mL for the resistant isolate. The EO (dillapiole 76.2%) was highly efficacious on phase L1. Due to the higher ICs obtained for the resistant isolate, it was raised the hypothesis that dillapiole may have a mechanism of action that resembles those of other anthelmintic compounds. We further review and discuss studies, especially those conducted in Brazil, that quantified the major constituents of P. aduncum-derived EO.

Resumo Este estudo avaliou a atividade anti-helmíntica in vitro do óleo essencial (OE) de Piper aduncum L. sobre ovos e larvas de Haemonchus contortus, verificando se um isolado resistente (Embrapa2010), apresentaria o mesmo comportamento que um sensível (McMaster). O OE foi obtido por arraste a vapor e analisado por cromatografia para identificação dos constituintes. O óleo foi avaliado nas concentrações de 12,5 a 0,02 mg/mL no Teste de eclosão dos ovos (TEO) e nas concentrações de 3,12 a 0,01 mg/mL no Teste de desenvolvimento larvar (TDL). As concentrações inibitórias (CI) foram determinadas pelo procedimento Probit do SAS e as diferenças estatísticas geradas pela ANOVA seguida pelo teste de Tukey. Para o isolado sensível obteve-se CI50 de 5,72 mg/mL no TEO. No TDL o óleo apresentou CI50 e CI90 de 0,10 mg/mL e 0,34 mg/mL para o isolado sensível e 0,22 mg/mL e 0,51 mg/mL para o resistente, respectivamente. Demonstrou-se que o OE (dilapiol 76,2%) teve alta eficácia sobre a fase L1. Devido às elevadas CIs obtidas para o isolado resistente, levantou-se a hipótese de que o dilapiol talvez possua um mecanismo de ação semelhante a algum grupo anti-helmíntico. O artigo faz uma revisão e discute estudos de quantificação dos constituintes majoritários do OE de P. aduncum, destacando os realizados no Brasil.

Antibacterial activity of the Piper aduncum oil and dillapiole, its main constituent, against multidrug-resistant strains / Actividad antibacteriana del aceite de Piper aduncum y dillapiole, su componente principal, frente a cepas resistentes a múltiples fármacos

The study aimed to evaluate the bactericidal activity of oil essential and dillapiole from P. aduncum against standard and multidrug-resistant strains of Staphylococcus spp. The oil showed antimicrobial action against these strains, but better results were obtained for the standards strains of S. epidermidis and S. aureus, with MIC of 250 and 500 ug/mL, respectively. Dillapiolle was less effective than the oil against the same standard and multi-drug resistant strains (MIC =1000 ug/mL). However, when dillapiolle was tested in combination with myristicin, another component of the oil, it increased its bactericidal activity and showed a synergistic action...

El objetivo del estudio fue evaluar la actividad bactericida de los aceites esenciales y dillapiole de P. aduncum contra cepas estándar y multirresistentes de Staphylococcus spp. El aceite mostró acción antimicrobiana frente a estas cepas, pero se obtuvo mejores resultados para las cepas de S. epidermidis y S. aureus, con MIC de 250 y 500 ug/ml, respectivamente. Dillapiolle fue menos eficaz que el aceite contra cepas estándar y multirresistentes (MIC = 1000 ug/ml). Sin embargo, cuando dillapiolle fue probado en combinación con la miristicina, otro componente del aceite, que aumentó su actividad bactericida y mostró una acción sinérgica...

Age-dependent changes from allylphenol to prenylated benzoic acid production in Piper gaudichaudianum Kunth.

HPLC-DAD and principal component analysis (PCA) of the (1)H NMR spectrum of crude plant extracts showed high chemical variability among seedlings and adult organs of Piper gaudichaudianum. While gaudichaudianic acid was the major compound in the adult leaves, apiole and dillapiole were the major compounds in their seedling leaves. By the 15th month of seedling growth, the levels of apiole and dillapiole decreased and gaudichaudianic acid appeared along with two compounds, biosynthetically related to gaudichaudianic acid.

Cytotoxic effects of dillapiole on MDA-MB-231 cells involve the induction of apoptosis through the mitochondrial pathway by inducing an oxidative stress while altering the cytoskeleton network.

Breast cancer is the world's leading cause of death among women. This situation imposes an urgent development of more selective and less toxic agents. The use of natural molecular fingerprints as sources for new bioactive chemical entities has proven to be a quite promising and efficient method. Here, we have demonstrated for the first time that dillapiole has broad cytotoxic effects against a variety tumor cells. For instance, we found that it can act as a pro-oxidant compound through the induction of reactive oxygen species (ROS) release in MDA-MB-231 cells. We also demonstrated that dillapiole exhibits anti-proliferative properties, arresting cells at the G0/G1 phase and its antimigration effects can be associated with the disruption of actin filaments, which in turn can prevent tumor cell proliferation. Molecular modeling studies corroborated the biological findings and suggested that dillapiole may present a good pharmacokinetic profile, mainly because its hydrophobic character, which can facilitate its diffusion through tumor cell membranes. All these findings support the fact that dillapiole is a promising anticancer agent.

Rendimento e composição do óleo essencial de Piper aduncum L. cultivado em Manaus, AM, em função da densidade de plantas e épocas de corte / Yield and composition of the essential oil from Piper aduncum L, cultivated in Manaus, AM, regarding plant density and cutting period

A pimenta-de-macaco (Piper aduncum L. - Piperaceae) é uma espécie usada na medicina popular em quase todo o Brasil e apresenta alto teor de óleo essencial (2,5 a 4,0%) rico em dilapiol - éter fenílico com ação inseticida. O presente trabalho teve como objetivo contribuir para o sistema de cultivo da espécie P. aduncum nas condições de Manaus - AM. O experimento foi realizado na área experimental da Embrapa Amazônia Ocidental. O delineamento utilizado foi o de blocos casualizados com quatro repetições, cada repetição com quatro plantas na área útil. Sete densidades de plantas (0,5 m x 0,5 m; 1,0 m x 0,5 m; 1,0 m x 1,0 m; 1,5 m x 1,0 m; 1,5 m x 1,5 m; 2,0 m x 1,5 m e 2,0 m x 2,0 m, E1 a E7, respectivamente) e três épocas de corte (C1 - 6 meses após transplantio; C2 - rebroto de C1 aos 6 meses - plantas com idade de 12 meses, no campo; C3 - rebroto de C2 aos 6 meses - plantas com 18 meses, no campo), portanto em esquema fatorial. Foi observado que o segundo corte (C2) apresentou maior produção, no espaçamento E4. Não houve diferença estatística para o teor de óleo essencial em C1, independente da densidade de plantas. Com relação ao rendimento (produção) em óleo essencial, o melhor foi observado com o corte C2, nos espaçamentos E3 e E4, com 76,0 kg/ha e 88 kg/ha, respectivamente, contendo até 85,6% de dilapiol no óleo. Assim, a melhor condição de cultivo nas condições de teste foi observada para o período de corte C2 e espaçamentos de 1,0 m x 1,0 m, e de 1,0 m x 1,5 m.

Pimenta-de-macaco (monkey pepper - Piper aduncum L. - Piperaceae) is a species used in folk medicine throughout Brazil, containing an essential oil (2.5 to 4.0% yield) rich in dillapiole, an arylpropanoid with insecticide action. The objective of this research was to improve the cultivation system of P. aduncum in local conditions of Manaus, AM. The study was conducted in the experimental fields of Embrapa Western Amazon, and used randomized blocks with four replicates, each replicate with four plants in its testing area. Seven plant densities (0.5 m x 0.5 m; 1.0 m x 0.5 m; 1.0 m x 1.0 m; 1.5 m x 1.0 m; 1.5 m x 1.5 m; 1.5 m 2.0 m; 2.0 m x 2.0 m, E1 to E7 respectively) and three cutting periods (C1 - 6 months after transplanting, C2 - regrowth of C1 after 6 months (plants with 12 months in the field) and C3 - regrowth of C2 after 6 months (plants with 18 months in the field), therefore in a factorial scheme. A larger production was observed in the second cutting (C2) in the plant density E4. There were no statistical differences in the essential oil content in C1, regardless of densities. In relation to the essential oil yield, the best result was observed for the cutting period C2 in the E3 and E4 densities, 76.0 kg/ha and 88 kg/ha, respectively, with a dillapiole content of up to 85.6%. Therefore, the best cultivation condition according to the test was observed for the cutting after 12 months of transplantation and plant density of 1.0 m x 1.0 m or 1.5 m x 1.5 m.