Impact of dietary leaf extracts of Black pepper Piper nigrum L on the growth, hematological and immunological parameters of Labeo rohita (Hamilton, 1822) cultured in glass aquaria / Impacto de extratos de folhas da dieta de pimenta-do-reino Piper nigrum L sobre o crescimento, parâmetros hematológicos e imunológicos de Labeo rohita (Hamilton, 1822) cultivado em aquários de vidro

A study was conducted to evaluate the effect of Piper nigrum (black pepper) leaf extract on on the growth performance, proximate composition, hematological parameters, and immune response of Labeo rohita fingerlings with an average weight of 22.14 ± 0.98g. After acclimation for two weeks, fish (n=25) were randomly selected and placed in four glass aquaria (T0, T1, T2 and T3) at constant water temperature (30.0 ± 1.0 °C), pH (7.50 ± 0.5) and total hardness (200 ± 2.0 mgL,-¹) for a period of 12 weeks, with three replicates each. Fish were fed with P. nigrum leaf extract supplemented feed @ 0.0%, 1.0%, 2.0% and 3.0% in T0, T1, T2 and T3, respectively. At the end of experiment, five fish were randomly selected from each aquaria for proximate composition, gut and skin microbial load, hematological parameters. Total proteins, albumins, and globulins were also recorded to evaluate immunological memory. The result revealed that fish in T2 showed better growth performance with an average weight gain of 56.11 ± 0.51 g. Thus, it had been concluded that Piper nigrum, a medicinal plant, can also be used to enhance the growth performance and immune response of Labeo rohita as attractive alternatives against antibiotics and vaccines and has shown no negative side effects on fish health as well as on its environment.

Um estudo foi conduzido para avaliar o efeito do extrato da folha de Piper nigrum (pimenta-do-reino) sobre o desempenho de crescimento, composição centesimal, parâmetros hematológicos e resposta imune de alevinos de Labeo rohita com peso médio de 22,14 ± 0,98g. Após aclimatação por duas semanas, os peixes (n = 25) foram selecionados aleatoriamente e colocados em quatro aquários de vidro (T0, T1, T2 e T3) em temperatura constante da água (30,0 ± 1,0 °C), pH (7,50 ± 0,5) e dureza total (200 ± 2,0 mgL-¹) por um período de 12 semanas, com três repetições cada. Os peixes foram alimentados com ração suplementada com extrato de folha de P. nigrum @ 0,0%, 1,0%, 2,0% e 3,0% em T0, T1, T2 e T3, respectivamente. Ao final do experimento, cinco peixes foram selecionados aleatoriamente de cada aquário para composição centesimal, carga microbiana intestinal e cutânea e parâmetros hematológicos. Proteínas totais, albuminas e globulinas também foram registradas para avaliar a memória imunológica. O resultado revelou que os peixes em T2 apresentaram melhor desempenho de crescimento com ganho de peso médio de 56,11 ± 0,51 g. Assim, concluiu-se que Piper nigrum, uma planta medicinal, também pode ser usado para melhorar o desempenho de crescimento e resposta imunológica de Labeo rohita como alternativas atraentes contra antibióticos e vacinas e não mostrou efeitos colaterais negativos na saúde dos peixes, bem como sobre seu ambiente.

Volatile chemical composition of Piper sancti-felicis Trel essential oil and its biocidal action against Tribolium castaneum (Herbst)

This research assessed the fumigant activity of the essential oil from Piper sancti-felicis Trel and five of its components on the Tribolium castaneum (Herbst) biological model. Hydrodistillation was used for extraction of the essential oil, with separation and identification of the compounds through gas chromatography coupled to mass spectrometry (GC-MS). The fumigant was evaluated through gas dispersion on the T. castaneum. The majority compounds found in the EO were b-nerolidol (15.4%), 3-carene (14.9%), p-cymene (9.1%), spathulenol (8.2%), a-cubebene (6.2%) and calamenene (5.2%). Piper sancti-felicis displayed fumigant activity with a LC50 = 108.5 & 956;g L-1 air, and other individual monoterpenes tested such as & 945;-terpinolene (LC50 = 110.1 & 956;g L-1 air), p-cymene (LC50 = 120.3 & 956;g L-1 air), 3-carene (LC50 = 130.6 & 956;g L-1 air), (R) -limonene (CL50 = 189.6 & 956;g L-1 air), and a-pinene (LC50 = 213.1 & 956;g L-1 air), were significantly less toxic than methyl pyrimiphos used as a positive control, CL50 = 87.4 & 956;g L-1 air. The essential oil of P. sancti-felicis can be considered as a natural source of biocides.

Phytochemistry and anti-inflammatory activities of Piper kadsura (Choisy) Ohwi ­ a review / Actividades fitoquímicas y antiinflamatorias de Piper kadsura (Choisy) Ohwi - una revisión

Piper kadsura (Choisy) Ohwi which belongs to the family Piperaceae, is a well-known medicinal plant possessing high medicinal and various therapeutic properties. It is widely used in traditional Chinese medicine for the treatment of asthma and rheumatic arthritis. Numerous studies on this species have also corroborated the significant anti-inflammatory potential of its extracts and secondary metabolites. The main chemical constituents which have been isolated and identified from P. kadsura are lignans and neolignans, which possess anti-inflammatory activities. The present article aims to provide a review of the studies done on the phytochemistry and antiinflammatory activities of P. kadsura. The scientific journals for this brief literature review were from electronic sources, such as Science Direct, PubMed, Google Scholar, Scopus, and Web of Science. This review is expected to draw the attention of the medical professionals and the general public towards P. kadsura and to open the door for detailed research in the future.

Piper kadsura (Choisy) Ohwi, perteneciente a la familia Piperaceae, es una planta medicinal conocida que posee importantes propiedades medicinales y diversas propiedades terapéuticas. Es ampliamente utilizada en la medicina tradicional china para el tratamiento del asma y la artritis reumática. Numerosos estudios sobre esta especie también han corroborado el destacado potencial antiinflamatorio de sus extractos y metabolitos secundarios. Los principales componentes químicos que se han aislado e identificado de P. kadsura son los lignanos y los neolignanos, que poseen actividades antiinflamatorias. El presente artículo tiene como objetivo proporcionar una revisión de los estudios realizados sobre las actividades fitoquímicas y antiinflamatorias de P. kadsura. Las revistas científicas para esta breve revisión de literatura fueron de fuentes electrónicas, como Science Direct, PubMed, Google Scholar, Scopus y Web of Science. Se espera que esta revisión atraiga la atención de los profesionales médicos y el público en general respecto de P. kadsura y abra la puerta a una investigación detallada en el futuro.

Larvicidal activity of piperovatine and dichloromethane extract from Piper corcovadensis roots against mosquitoes Aedes aegypti L / Actividad larvicida de piperovatina y extracto de diclorometano de las raíces de Piper corcovadensis contra mosquitos vectores del dengue Aedes aegypti L

The research of new substances capable of controlling the Aedes aegypti mosquito is urgent due to the increase in the transmission of the diseases such as dengue, chikungunya and Zika virus by the vector. Thus, the aim of this study was to evaluate the larvicidal activity of crude extract of Piper corcovadensis roots, a native plant from Brazil, and of the isolated compound piperovatine against larvae of A. aegypti by the larval immersion test. The lethal concentration that killed 50% (LC50) and 99% (LC99) of larvae was determined by Probit analysis. The results indicated high larvicidal activity on A. aegypti larvae for crude extract of Piper corcovadensis roots with LC50 of 4.86 µg/mL and LC99 of 15.50 µg/mL and piperovatine with LC50 of 17.78 µg/mL and LC99 of 48.55 µg/mL. This work opens new perspectives to the development of future products with crude extract of Piper corcovadensis roots and piperovatine that can be applied to mosquito control.

La investigación de nuevas sustancias capaces de controlar el mosquito Aedes aegypti es urgente debido al aumento en la transmisión de enfermedades como el dengue, el chikungunya y el virus Zika por el vector. Por lo tanto, el objetivo de este estudio fue evaluar la actividad larvicida del extracto crudo de las raíces de Piper corcovadensis, una planta nativa de Brasil, y del compuesto aislado piperovatine contra larvas de A. aegypti mediante la prueba de inmersión larvaria. La concentración letal que mató al 50% (LC50) y al 99% (LC99) de larvas se determinó mediante análisis Probit. Los resultados indicaron una alta actividad larvicida en larvas de A. aegypti para extracto crudo de las raíces de Piper corcovadensis con LC50 de 4.86 µg/mL y LC99 de 15.50 µg/mL y piperovatine con LC50 de 17.78 µg/mL y LC99 de 48.55 µg/mL. Este trabajo abre nuevas perspectivas para el desarrollo de futuros productos con extracto crudo de las raíces de Piper corcovadensis y piperovatine que pueden aplicarse al control de mosquitos.

Aporphine alkaloids from Piper erecticaule and acetylcholinesterase inhibitory activity / Alcaloides de aporfina obtenida de Piper erecticaule y actividad inhibitoria de la acetilcolinesterasa

Chemical constituents and biological activities of the aerial parts of Piper erecticaule C.DC. have been studied for the first time. Fractionation and purification of the extracts afforded aristolactam AII (1), aristolactam BII (2), piperolactam A (3), piperolactam C (4), piperolactam D (5), together with terpenoids of ß-sitosterol, ß-sitostenone, taraxerol, and lupeol. The structures of these compounds were obtained by analysis of their spectroscopic data, as well as the comparison with that of reported data. Acetylcholinesterase inhibitory activity revealed that compounds 1 and 3 showed strong AChE inhibitory effects with the percentage inhibition of 75.8% and 74.8%, respectively.

Se estudiaron por primera vez los constituyentes químicos y actividad biológica de las partes aéreas de Piper erecticaule C.DC. El fraccionamiento y la purificación de los extractos proporcionaron aristolactama AII (1), aristolactama BII (2), piperolactama A (3), piperolactama C (4), piperolactama D (5), junto con terpenoides de ß-sitosterol, ß-sitostenona, taraxerol, y el lupeol. Las estructuras de estos compuestos se obtuvieron mediante el análisis de sus datos espectroscópicos, así como mediante la comparación con datos ya informados. La actividad inhibidora de la acetilcolinesterasa reveló que los compuestos 1 y 3 mostraron un potente efecto inhibidor de la AChE con un porcentaje de inhibición del 75.8% y 74.8%, respectivamente.

First record of the chemical composition of essential oil of Piper bellidifolium, Piper durilignum, Piper acutilimbum and Piper consanguineum from the Brazilian Amazon forest / Primeiro registro da composição química de óleos essenciais de Piper bellidifolium, Piper durilignum, Piper acutilimbum e Piperconsanguineumda Floresta Amazônica no Brasil

Piper bellidifolium, Piper durilignum, Piper acutilimbum and Piper consanguineum are bushes that occur in the Amazon and are morphologically similar. With the aim of analyzing the chemical profile of the volatile constituents of these species, essential oils from the leaves were obtained through steam distillation and analyzed using gas chromatography-flame ionization detection (GC-FID) and gas chromatograph coupled to a mass spectrometer (GC-MS). The chemical analysis enabled the identification of 95 compounds representing 96.3 ± 0.6% of the P. bellidifolium oil, 95.5 ± 0.71% of the P. durilignum oil, 98.0 ± 1.0% of the P. acutilimbum oil and 96.1 ± 2.1% of the P. consanguineum oil. Although sesquiterpenes were the predominant chemical class in the oils of the four species, qualitative and quantitative differences were found in their chemical composition. The major constituents were (E)-nerolidol (20.3 ± 0.4%) in the P. bellidifolium oil, germacrene D (11.1 ± 0.3%) in the P. durilignum oil, and -eudesmol in both the P. consanguineum (18.6 ± 0.5%) and P. acutilimbum (7.5 ± 0.4%) oils. Despite their morphological similarity, a principal component analysis (PCA) of the GC-MS data clearly separated the four species according to the chemical profile of the essential oil extracted from their leaves.

Piper bellidifolium, Piper durilignum, Piper acutilimbum e Piper consanguineum são arbustos que ocorrem na Amazônia e são morfologicamente similares. Com o intuito de analisar o perfil químico dos constituintes voláteis dessas espécies, óleos essenciais das folhas foram obtidos por hidrodestilação e analisados por cromatografia gasosa - detector por ionização de chama (CG-FID) e cromatografia gasosa acoplada a espectrometria de massa (CG-EM). A análise química permitiu identificar 95 compostos, representando 96.3 ± 0.6% do óleo de P. bellidifolium; 95.5 ± 0.71% de P. durilignum; 98.0 ± 1.0% de P. acutilimbum e 96.1 ± 2.1% de P. consanguineum. Apesar dos óleos das quatro espécies terem sesquiterpeno como classe química predominante, diferenças qualitativas e quantitativas em sua composição química foram observadas. Os principais componentes encontrados foram: (E)-nerolidol (20.3 ± 0.4%) em P. bellidifolium; germacreno D (11.1 ± 0.3%) em P. durilignum; e -eudesmol nos óleos de P. consanguineum (18.6 ± 0.5%) e P. acutilimbum (7.5 ± 0.4%). Apesar da similaridade morfológica entre as espécies, uma análise de componentes principais (PCA) dos dados de CG-EM claramente separou as quatro espécies quanto ao perfil químico do óleo essencial extríado de suas folhas.

Antiplasmodial and antileishmanial activities of compounds from Piper tuberculatum Jacq fruits

Abstract INTRODUCTION This study assessed the activity of compounds from Piper tuberculatum against Plasmodium falciparum and Leishmania guyanensis. METHODS The effects of compounds from P. tuberculatum fruits on P. falciparum and L. guyanensis promastigote growth in vitro were determined. Hemolytic action and cytotoxicity in HepG2 and J774 cells were measured. RESULTS Three compounds showed strong antiplasmodial activity and one compound showed strong antileishmanial activity. Two compounds were non-toxic to HepG2 cells and all were toxic to J774 cells. The compounds showed no hemolytic activity. CONCLUSIONS The tested compounds from P. tuberculatum exhibited antiparasitic and cytotoxic effects.

Proteomic profile of Piper tuberculatum (Piperaceae) / Perfil proteômico e metabólico de Piper tuberculatum (Piperaceae)

Abstract Piper tuberculatum (Piperaceae) is a species that accumulates especially amides as secondary metabolites and several biological activities was previously reported. In this article, we report a proteomic study of P. tuberculatum. Bidimensional electrophoresis (2D SDS-PAGE) and mass spectrometry (ESI-Q-TOF) were used in this study. Over a hundred spots and various peptides were identified in this species and the putative functions of these peptides related to defense mechanism as biotic and abiotic stress were assigned. The information presented extend the range of molecular information of P. tuberculatum.

Resumo Piper tuberculatum (Piperaceae) é uma espécie que acumula especialmente amidas como metabólitos secundários e diversas atividades biológicas dessa espécie foram relatadas anteriormente. No presente artigo, relatamos um estudo proteômico dessa espécie. Eletroforese bidimensional (2D SDS-PAGE) e espectrometria de massas (ESI-Q-TOF) foram utilizadas nesse estudos. Mais de cem spots e vários peptídeos foram identificados nesta espécie e as funções putativas desses peptídeos relacionadas a mecanismo de defesa como estresse biótico e abiótico foram atribuídos. As informações apresentadas ampliam a gama de informações moleculares dessa espécie.

Biological Activity of Piper aduncum extracts on Anticarsia gemmatalis (Hübner) (Lepidoptera: Erebidae) and Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae)

ABSTRACT Piper aduncum found naturally in the Amazon and southeastern Brazil, is known for its secondary metabolites that have activity on insects. Anticarsia gemmatalis and Spodoptera frugiperda are among the major insect pests associated with agricultural production. This research evaluated the biological activity of hexane, ethyl acetate, and ethanol extracts of P. aduncum leaves on mortality and duration of larval and pupal periods, as well as weight, width, and length of A. gemmatalis and S. frugiperda pupae. The mortality of A. gemmatalis larvae in trials with P. aduncum extracts were 93.3% (hexane) and 90% (ethyl acetate), estimating LC50 of 6.35 and 5.79 mg/mL, respectively. Mortality in S. frugiperda submitted to the hexane extract ranged from 3.33% to 96.66% (LC50 of 8.22 mg/mL). The ethanol extract induced low mortality (3.33% to 23.33%). The P. aduncum extracts did not affect the development of S. frugiperda pupae. In A. gemmatalis differences in weight and length occurred. The chemical characterization was by GC-MS, which revealed that the major constituent in the hexane extract of P. aduncum was apiol (90.7%). P. aduncum extracts are important and promising components to manage A. gemmatalis and S. frugiperda, which cause extensive production losses.

Linalool, a Piper aduncum essential oil component, has selective activity against Trypanosoma cruzi trypomastigote forms at 4°C

BACKGROUND Recent studies showed that essential oils from different pepper species (Piper spp.) have promising leishmanicidal and trypanocidal activities. OBJECTIVES In search for natural compounds against Trypanosoma cruzi, different forms of the parasite were incubated for 24 h at 28ºC or 4ºC with Piper aduncum essential oil (PaEO) or its main constituents linalool and nerolidol. METHODS PaEO chemical composition was obtained by GC-MS. Drug activity assays were based on cell counting, MTT data or infection index values. The effect of PaEO on the T. cruzi cell cycle and mitochondrial membrane potential was evaluated by flow cytometry. FINDINGS PaEO was effective against cell-derived (IC50/24 h: 2.8 μg/mL) and metacyclic (IC50/24 h: 12.1 μg/mL) trypomastigotes, as well as intracellular amastigotes (IC50/24 h: 9 μg/mL). At 4ºC - the temperature of red blood cells (RBCs) storage in blood banks - cell-derived trypomastigotes were more sensitive to PaEO (IC50/24 h = 3.8 μg/mL) than to gentian violet (IC50/24 h = 24.7 mg/mL). Cytotoxicity assays using Vero cells (37ºC) and RBCs (4ºC) showed that PaEO has increased selectivity for cell-derived trypomastigotes. Flow cytometry analysis showed that PaEO does not affect the cell cycle of T. cruzi epimastigotes, but decreases their mitochondrial membrane potential. GC-MS data identified nerolidol and linalool as major components of PaEO, and linalool had trypanocidal effect (IC50/24 h: 306 ng/mL) at 4ºC. MAIN CONCLUSION The trypanocidal effect of PaEO is likely due to the presence of linalool, which may represent an interesting candidate for use in the treatment of potentially contaminated RBCs bags at low temperature.

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.

Determinación del efecto cicatrizante de Piper aduncum (matico) en fibroblastos humanos / Determination of the healing effect of Piper aduncum (spiked pepper or matico) on human fibroblasts

RESUMEN Objetivos. Evaluar el efecto cicatrizante del extracto hidroetanólico de Piper aduncum, en una línea celular de fibroblastos Dermales Adultos Humanos (hDFa). Materiales y métodos. El extracto se obtuvo mediante extracción sólido-líquido, fue concentrado y liofilizado. Se purificaron las proteínas del extracto mediante cromatografía líquida de alta eficacia de fase reversa (RP-HPLC); las proteínas fueron identificadas por espectrometría de masas en tándem de péptidos trípticos y se analizaron por MALDI-TOF-TOF en un espectrómetro de masa ABSciex4800. Los valores de concentración efectiva media (EC50), concentración inhibitoria media (IC50), y el porcentaje de proliferación celular; fueron determinados por ensayos con sales de tetrazolio (MTT) . La migración celular se evaluó mediante la "técnica de rayado" . Se analizó la expresión de factores de crecimiento mediante el ensayo de reacción en cadena de la polimerasa con transcriptasa reversa a tiempo real (RT- qPCR). Resultados. La línea hDFa evidenció un IC50 de 200 µg/mL con el extracto, el valor de EC50 fue 103,5 µg/mL. En el ensayo de proliferación, la proteína K2; mostró mayor actividad en la proliferación respecto de otros tratamientos (1 µg/mL). En el ensayo de migración de fibroblastos, la proteína K2 mostró mayor actividad (50 µg/mL). La expresión relativa del factor de crecimiento derivado de plaquetas (PDGF) se incrementó 8,6 veces respecto al control, en presencia de la proteína K2. Conclusiones. El extracto hidroetanólico, de Piper aduncum, así como las proteínas que contiene, incrementaron la proliferación y migración de fibroblastos dermales humanos (hDFa); así mismo, aumentaron la expresión de factores de crecimiento que intervienen en el proceso de cicatrización.

ABSTRACT Objectives. To evaluate the healing effect of a Piper aduncum ethanol-water extract on an adult human dermal fibroblast cell line (hDFa). Materials and Methods. After obtaining the extract via solid-liquid extraction, concentration, and lyophilization, extract proteins were purified using reverse phase high-performance liquid chromatography, identified using tandem mass spectrometry of tryptic peptides, and analyzed using MALDI-TOF-TOF on an ABSciex4800 mass spectrometer. Half maximum effective concentration values (EC50), half maximum inhibiting concentration (IC50), and percentages of cell proliferation were determined using tetrazolium salt assays. Cell migration was evaluated using a "scratch assay". Growth factor expression in cells was analyzed via quantitative real-time reverse transcription polymerase chain reaction. Results. Against the hDFa cell line, the extract had an IC50 of 200 μg/mL and EC50 of 103.5 µg/mL. In the proliferation assay, protein K2 (obtained from the extract) exhibited increased proliferative activity relative to other treatments (1 µg/mL); this agent also exhibited increased activity (50 µg/mL) in the fibroblast migration assay.Furthermore, the relative expression of platelet-derived growth factor increased by 8.6-fold in the presence of K2 protein relative to the control. Conclusions. The hydroethanolic extract of Piper aduncum and its component proteins increased the proliferation and migration of hDFa and increased the expression of growth factors involved in the healing process.

Concentraciones de alcaloides, glucósidos cianogénicos, polifenoles y saponinas en plantas medicinales seleccionadas en Ecuador y su relación con la toxicidad aguda contra Artemia salina / Concentrations of alkaloids, cyanogenic glycosides, polyphenols and saponins in selected medicinal plants from Ecuador and their relationship with acute toxicity against Artemia salina

ResumenEntre los principales compuestos químicos sintetizados por las plantas, pero considerados no esenciales para su metabolismo básico, están los alcaloides, los polifenoles, los glucósidos cianogénicos y las saponinas que tienen diversas funciones en las plantas y reconocidas propiedades medicinales y farmacológicas. En esta investigación se determinaron las concentraciones de los mencionados metabolitos secundarios en los extractos de las hojas de las plantas medicinales Taraxacum officinale, Parthenium hysterophorus, Artemisia absinthium, Cnidoscolus aconitifolius y Piper carpunya y se relacionaron con la toxicidad aguda contra Artemia salina. En cada bioensayo con A. salina se usaron los extractos alcohólicos de las hojas de las plantas seleccionadas a diferentes concentraciones, calculándose la proporción de organismos muertos y los CL50. Las concentraciones de alcaloides, fenoles totales, taninos, glucósidos cianogénicos y saponinas fueron determinadas mediante métodos espectrofotométricos. Este es el primer reporte de cuantificación de metabolitos secundarios en las plantas analizadas provenientes de Ecuador. T. officinale presentó las mayores concentraciones de fenoles (22.30 ± 0.23 mg/g) y taninos (11.70 ± 0.10 mg/g), C aconitifolius de glucósidos cianogénicos (5.02 ± 0.37 µg/g) y P. hysterophorus de saponinas (6.12 ± 0.02 mg/g). Las plantas evaluadas presentaron actividades hemolíticas dependiendo de las concentraciones de saponinas. Los valores de taninos determinados estuvieron entre 0.20 ± 0.01 y 11.70 ± 0.10 mg/g, por lo que no son adversos para su consumo. Aunque los valores de glucósidos cianogénicos son permisibles, es necesario monitorear la presencia de estos compuestos químicos en las plantas para minimizar problemas de salud. Los CL50 obtenidos oscilaron entre los valores 3.37 µg/mL, extremadamente letal o tóxica, para P. carpunya y 274.34 µg/mL, altamente tóxica, para T. officinale. De los análisis de correlaciones realizados a los resultados, se observó que los alcaloides favorecen de manera significativa (p<0.001) a la toxicidad aguda contra A. salina, mientras que a mayor contenido de polifenoles dicha toxicidad disminuye significativamente (p<0.001) el nivel de toxicidad de las plantas. Del análisis de componentes principales, se demuestra que las saponinas están en sinergia con los polifenoles para disminuir la toxicidad, pero tienen un efecto antagónico con los alcaloides y los glucósidos cianogénicos, lo cual evidencia que estos metabolitos secundarios presentan variabilidades en los mecanismos de acción contra A. salina, como compuestos citotóxicos. Estos resultados demuestran que las saponinas y los polifenoles pueden ser letales para A. salina a bajas concentraciones, evidenciando que este bioensayo permite evaluar extractos vegetales que contengan bajas concentraciones de compuestos con altas polaridades. La correspondencia significativamente positiva entre citoxicidad y concentración de los alcaloides, confirmada con el bioensayo de Artemia salina, puede ser útil para hallar fuentes promisorias de compuestos antitumorales y para evaluar los límites tolerables que no afecten otras células benignas. El contenido de metabolitos secundarios hallados en las plantas analizadas les atribuye un gran valor farmacológico.

Abstract:Alkaloids, polyphenols, cyanogenic glycosides and saponins are among the main chemical compounds synthesized by plants but not considered essential for their basic metabolism. These compounds have different functions in plants, and have been recognized with medicinal and pharmacological properties. In this research, concentrations of the mentioned secondary metabolites were determined in the medicinal plants Artemisia absinthium, Cnidoscolus aconitifolius, Parthenium hysterophorus, Piper carpunya and Taraxacum officinale, from Ecuador, and related with cytotoxic effects against Artemia salina. Alcoholic and aqueous extracts from leaves of these selected plants were prepared at different concentrations. To assess cytotoxicity of these extracts, different bioassays with A. salina were undertaken, and the mortality rates and LC50 were obtained. Besides, concentrations of alkaloids, cyanogenic glycosides, phenols, tannins and saponins were determined by spectrophotometric methods; this constituted the first report of quantification of secondary metabolites in the selected plants from Ecuador. T. officinale had the highest concentration of total phenols (22.30 ± 0.23 mg/g) and tannins (11.70 ± 0.10 mg/g), C. aconitifolius of cyanogenic glycosides (5.02 ± 0.37 µg/g) and P. hysterophorus of saponins (6.12 ± 0.02 mg/g). Tannins values obtained were not adverse to their consumption. Alcoholic and aqueous extracts of selected plants had hemolytic activity depending on the concentration of saponins. Although the values of cyanogenic glycosides were permissible, it was necessary to monitor the presence of this metabolite in plants to minimize health problems. LC50 values ranged from extremely toxic (3.37 µg/mL) to highly toxic (274.34 μg/mL), in P. carpunya and T. officinale, respectively. From correlation analysis, it was observed that increase values of alkaloids concentrations had highly significant (p<0.001) acute toxicity against A. salina, while at a higher polyphenol concentration the level of plants cytotoxicity decreased significantly (p<0.001). The results of principal component analysis showed that saponins apparently were in synergy with polyphenols to decrease cytotoxicity, but antagonize with alkaloids and cyanogenic glycosides, indicating that these secondary metabolites present variability in the mechanisms of action against A. salina, as cytotoxic compounds. These results also demonstrate that polyphenols and saponins can be lethal at low concentrations, demonstrating the potential of brine shrimp bioassay as a model to evaluate plant extracts containing low concentrations of chemical compounds with high polarities. The significant positive correlation between cytotoxicity and concentration of alkaloids confirmed by the bioassay of brine shrimp can be useful to identify promising sources of antitumor compounds, and to evaluate tolerable limits not affecting other benign cells. Contents of secondary metabolites found in the selected plants confer them great pharmacologic values. Rev. Biol. Trop. 64 (3): 1171-1184. Epub 2016 September 01.

Atividade antimicrobiana de produtos obtidos de espécies de Piper (Piperaceae) / Actividad antimicrobiana de productos obtenidos a partir de especies de Piper (Piperaceae) / Antimicrobial activity of products obtained from Piper species (Piperaceae)

Introdução: espécies do gênero Piper são encontradas nas regiões tropicais do globo e possuem inúmeras indicações terapêuticas. Objetivos: avaliar a atividade antimicrobiana de extratos e frações de quatro espécies de Piper, além de dois flavonoides isolados de Piper montealegreanum e determinar a concentração inibitória mínima dos produtos ativos. Métodos: o material vegetal foi coletado em diferentes regiões do estado da Paraíba e na cidade de Belém (Pará) submetido à extração com etanol e partição com solventes em grau crescente de polaridade. Os flavonoides foram isolados por meio de técnicas cromatográficas. A avaliação da atividade antimicrobiana foi realizada pelo método de disco-difusão frente a três espécies bacterianas: Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853 e três espécies fúngicas: Candida albicans ATCC 76645, Candida tropicalis ATCC 13803 e Candida glabrata ATCC 15545. Resultados: quase metade dos produtos testados foi capaz de inibir o crescimento de S. aureus com halos de inibição variando entre 8-16 mm. O extrato etanólico bruto e as frações hexânica e CHCl3 das folhas de P. mollicomum mostraram atividade antibacteriana frente as três linhagens testadas. As frações hexânicas de P. arboreum, de P. caldense e de P. mollicomum apresentaram atividade antifúngica, enfatizando a formação do halo de inibição de 28 mm para a fração de P. mollicomum. Os dois flavonoides isolados de P. montealegreanum não apresentaram atividade antimicrobiana. Conclusões: os resultados preliminares deste estudo sugerem que alguns dos produtos obtidos das espécies de Piper são uma fonte promissora de compostos com atividade antimicrobiana(AU)

Introducción: el género Piper se encuentra en las regiones tropicales del mundo y numerosas indicaciones terapéuticas. Objetivos: evaluar la actividad antimicrobiana de extractos y fracciones de cuatro especies de Piper, además de dos flavonoides aislados de Piper montealegreanum y determinar la concentración inhibitoria mínima de los productos activos. Métodos: el material vegetal fue recolectado en diferentes regiones del estado de Paraíba y en Belém (Pará) y sometido a extracción con etanol y partición com disolventes en grado creciente de polaridad. Los flavonoides fueron aislados por medio de técnicas cromatográficas. La evaluación de la actividad antimicrobiana se realizó por el método de difusión en disco frente a tres especies bacterianas:Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853 y três especies de hongos: Candida albicans ATCC 76645, Candida tropicalis ATCC 13803 y Candida glabrata ATCC 15545. Resultados: casi la mitad de los productos ensayados fueron capaces de inhibir el crecimiento de S. aureus con zonas de inhibición entre 8-16 mm. Los extractos etanolicos crudo y fracciones hexano y CHCl3 de las hojas de P. mollicomum mostraron actividad antibacteriana contra todas las cepas probadas. Las fracciones hexano de P. arboreum, P. caldense y P. mollicomum mostraron actividad antifúngica, con énfasis en la zona de inhibición de 28 mm para la fracción de P. mollicomum. Los dos flavonoides aislados de P. montealegreanum no mostraron actividad antimicrobiana. Conclusiones: los resultados preliminares de este estudio sugieren que algunos de los productos obtenidos de especies de Piper son una fuente prometedora de compuestos con actividad antimicrobiana(AU)

Introduction: The species of the genus Piper are widely found in the tropical regions of the globe and have numerous therapeutic indications. Objective: To evaluate the antimicrobial activity of extracts and fractions of four species of Piper and two flavonoids isolated from Piper montealegreanum and determine the minimum inhibitory concentration of the active products. Methods: The plant material was collected in different regions of the state of Paraíba and in Belém (Pará) and subjected to extraction with ethanol and partition with solvents in increasing degree of polarity. Flavonoids were isolated through chromatographic techniques. The evaluation of antimicrobial activity was performed by disk diffusion method front the three bacterial species: Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853 and three fungi species: Candida albicans ATCC 76645, Candida tropicalis ATCC 13803 and Candida glabrata ATCC 15545. Results: Almost half of the products tested were able to inhibit the growth of S. aureus with inhibition zones between 8-16 mm. The crude ethanolic extract and hexane fractions and CHCl3 of P. mollicomum leaves showed antibacterial activity against all the tested strains. The hexane fractions of P. arboreum, P. caldense and P. mollicomum showed antifungal activity, emphasizing the formation of 28 mm inhibition zone for the fraction of P. mollicomum. The two flavonoids isolated from P. montealegreanum showed no antimicrobial activity. Conclusions: preliminary results suggest that some the products obtained from Piper species are a promising source of compounds with antimicrobial activity(AU)

Caracterização farmacognóstica de Piper arboreum var. arboreum e P. tuberculatum (Piperaceae) / Pharmacognostical characterization of Piper arboreum var. arboreum and P. tuberculatum (Piperaceae)

Piper arboreum Aubl. var. arboreum and P. tuberculatum Jacq. (Piperaceae), both known as "pimenta-longa", are frequently misidentified. Despite the fact that both species are sold as traditional phytotherapeutic products, there are no reports on their quality control. The aim of this study was to prepare and characterize leaf and stem herbal drugs of both species using standard methods aiming to establish parameters for botanical authentication and purity degree for their quality control. Size, shape, color, taste, texture, fracture surface and transection characteristics, and leaf venation patterns are macroscopic diagnostic characters useful to distinguish the herbal drugs when they are not powdered. The following characters are useful for diagnostic purposes of the powdered herbal drugs: cuticular and epicuticular wax patterns, types of trichomes, and wall thickening of the tracheary elements, and shape of the hilum of the starch grains. Emergences are key morphological markers for P. tuberculatum herbal drugs. Physicochemical parameter values were similar in herbal drugs of both species, except acid-insoluble ash value which was higher in the P. arboreum var. arboreum stem herbal drugs. Piper arboreum var. arboreum herbal drugs had the highest metal concentrations. Al, Cu, Mn and V concentrations were the highest, whereas Cd, Co and Hg concentrations were the lowest in the herbal drugs of both species. The results obtained can be used as quality control parameters for the herbal drugs of both species.

Piper arboreum Aubl. var. arboreum e P. tuberculatum Jacq. (Piperaceae), conhecidas como pimenta-longa, são comercializadas como produtos tradicionais fitoterápicos e, frequentemente, identificadas erroneamente, não havendo relatos sobre controle de qualidade dessas espécies. O objetivo do trabalho foi preparar e caracterizar drogas vegetais foliares e caulinares dessas espécies para estabelecer-lhes parâmetros de autenticidade botânica e grau de pureza para o controle de qualidade. Foram utilizados métodos padrões para o preparo e análises estrutural e físico-química das drogas vegetais. São caracteres diagnósticos macroscópicos para distinguir as drogas vegetais íntegras: tamanho, forma, cor, sabor, textura, características da superfície de fratura e da secção transversal e os padrões de venação foliar. São caracteres diagnósticos para distinguir as drogas vegetais pulverizadas: padrões de cutícula e cera epicuticular, tipos de tricoma e espessamento parietal dos elementos traqueais e a forma do hilo dos grãos de amido. Emergências são marcadores morfológicos chave para as drogas vegetais de P. tuberculatum. Os valores dos parâmetros físico-químicos foram similares nas drogas vegetais das espécies, exceto o de cinzas ácido-insolúveis que foi maior nas drogas vegetais caulinares de P. arboreum var. arboreum. As maiores concentrações de metais ocorreram nas drogas vegetais de P. arboreum var. arboreum. As concentrações mais elevadas foram de Al, Cu, Mn e V e as mais baixas de Cd, Co e Hg. Os resultados obtidos poderão ser utilizados como parâmetros de controle de qualidade para as drogas vegetais de ambas as espécies.

Composition of the essential oils from leaves of Piper lepturum Kunth (C.DC.) var. lepturum and Piper lepturum var. angustifolium (Miq.) Yunck. from Brasil / Composición de los aceites esenciales de hojas de Piper lepturum Kunth (C.DC.) var. lepturum y Pip

The essential oils of Brazilian Piper lepturum var. lepturum and Piper lepturum var. angustifolium (Piperaceae) were obtained by hydrodistillation and analyzed by flame-detector gas chromatography (GC) and gas chromatography coupled to mass spectrometry (GC/MS). According to GC and GC/MS analysis, the essential oils are mostly composed by sesquiterpenes hydrocarbons. beta-Guaiene (29.96 percent) was the principal component in the essential oil of P. lepturum var. lepturum and beta-Bisabolene (17.72 percent) was the principal components in the essential oil of P. lepturum var. angustifolium.

Los aceites esenciales de las especies brasileñas Piper lepturum var. lepturum y Piper lepturum var. angustifolium fueron obtenidos por hidrodestilación y analizados utilizando cromatografía gas líquido con detector de ionización de llama (CG) y cromatografía gas líquido acoplada a un detector de masas (CG/EM). De acuerdo con los análisis de CG y CG/EM, los aceites esenciales muestran como componente principal beta-Guaieno (29,96 por ciento) en el aceite esencial de P. lepturum var. lepturum y beta-Bisaboleno (17,71 por ciento) en el aceite esencial de P. lepturum var. angustifolium.

Estudio comparativo de de la composición volátil y las actividades biológicas del aceite esencial de Piper marginatum Jacq colombiano / Comparative study of volatile composition and biological activities of essential oil from Colombian Piper marginatum Jacq

Los objetivos de este trabajo fueron el estudio de la composición química volátil del aceite esencial (AE) de Piper marginatun Jacq (Piperaceae) obtenido de plantas colectadas en dos sitios geográficos colombianos: municipios de Acandí (Chocó) y Turbaco (Bolívar). Además, evaluar sus propiedades antioxidante, repelente y antialimentaria. AEs fueron aislados mediante la técnica de hidrodestilación e identificados por cromatografía de gases acoplada a un detector de espectrometría de masas (GC-MS). Los compuestos mayoritarios encontrados en P. marginatum colectado en Acandi fueron cis-p-anetol (46.3 por ciento), estragol (28.9 por ciento), safrol (9.0 por ciento), mientras que en P. marginatum de Turbaco fueron germacreno D (36.6 por ciento), beta-elemeno (12.6 por ciento), germacren-D-4-ol (5.4 por ciento). La actividad antioxidante de los AEs se determinó por el método del radical DPPH.. Los porcentajes de inhibición de DPPH. del aceite esencial de P marginatum de Turbaco y Acandí fueron del 90.0 por ciento y 73.7 por ciento, respectivamente. El AE procedente de Turbaco presentó la mayor actividad repelente frente al gorgojo Tribolium castaneum, a una concentración de 0.1 uL/cm2 a 2 y 4 horas de exposición (85 por ciento y 76 por ciento, respectivamente). El AE de P. marginatum de Turbaco fue más activo contra las larvas del Spodoptera littoralis (Porcentaje de inhibición de la alimentación, porcentaje FR = 80,6) y se observó una elevada inhibición de asentamiento de Myzus persicae y Rhopalosiphum padi (porcentaje SI = 87,9% y 86,4%).

The objectives of this work were to study the volatile chemical composition of essential oil (EO) from Piper marginatun Jacq (Piperaceae), obtained from plants collected in two Colombian geographic locations: municipalities Acandí (Chocó) and Turbaco (Bolívar). Also, evaluate their antioxidant, repellent and antifeedant properties. EOs were isolated by hydrodistillation technique and identified by gas chromatography coupled to a mass spectrometric detector (GC-MS). The major compounds found in P. marginatum collected in Acandi were cis-p-anethole (46.3 percent), estragole (28.9 percent), safrole (9.0 percent), while in P. marginatum from Turbaco were germacrene D (36.6 percent), beta- elemene (12.6 percent), germacren-D-4-ol (5.4 percent). The antioxidant activity of the EOs were determined using the method of DPPH radical. The percentages of inhibition of DPPH. of P marginatum from Turbaco and Acandí were 90.0 percent and 73.7 percent, respectively. The EO from Turbaco had the highest repellent activity against Tribolium castaneum, at a concentration of 0.1 uL /cm2 at 2 and 4 hours of exposure (85 percent and 76 percent respectively). P. marginatum from Turbaco was more active against Spodoptera littoralis larvae (Percent feeding inhibition, FI= 80.6 percent) and a high Percent settling inhibition of Myzus persicae and Rhopalosiphum padi (SI = 87.9% and 86,4%).

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...

Cell cycle inhibitory activity of Piper longum against A549 cell line and its protective effect against metal-induced toxicity in rats.

Anticancer potential of Piper longum fruit against human cancer cell lines (DU-145 prostate, A549 lung, THP-1 leukemia, IGR-OVI-1 ovary and MCF-7 breast) as well as its in vitro and in vivo biochemical efficacy in AlCl3-induced hepatotoxicity were evaluated in the rats. Dried samples were extracted with several solvents using soxhlet apparatus. Flavonoid content in chloroform, benzene, ethyl alcohol and aqueous extracts of fruit was 19, 14, 12 and 11 μg quercetin equivalent/mg of sample, respectively. Hexane extracts exhibited 90-92% cytotoxicity against most of the test cell lines (A549, THP-1, IGR-OVI-1 and MCF-7), while benzene extract displayed 84-87% cytotoxicity against MCF-7, IGR-OV-1 and THP-1 cell lines. Among extracts, hexane, benzene and acetone extracts demonstrated considerable cytotoxicity (91-95%) against A549 (lung cancer) cell line in Sulforhodamine B dye (SRB) assay. Cell cycle analysis revealed that hexane, benzene and acetone extracts produced 41, 63 and 43% sub-G1 DNA fraction, demonstrating cell cycle inhibitory potential of these extracts against A549 cell line. Chloroform, ethyl alcohol and aqueous extracts displayed 71, 64 and 65% membrane protective activity, respectively in lipid peroxidation inhibition assay. P. longum fruit extracts also ameliorated AlCl3-induced hepatotoxicity, as indicated by alterations observed in serum enzymes ALP, SGOT and SGPT activity, as well as creatinine and bilirubin contents. In conclusion, study established the cytotoxic and hepatoprotective activity in P. longum extracts.

Chemical composition of Piper stylosum Miq. and Piper ribesioides Wall.essential oils, and their antioxidant, antimicrobial and tyrosinaseinhibition activities / Composición química de los aceites esenciales de Piper stylosum Miq. y Piper ribosoides Wall. y sus actividadesantioxidantes, antimicrobiana y de inhibición de la tirosinasa

Chemical compositions of P. stylosum and P. ribesioides essential oils, and their antioxidant, antimicrobial and tyrosinase inhibition activities were determined. GC and GC–MS analysis of essential oils from leaves and stems of P. stylosum resulted in the identification of 50 (89.2 percent) and 45 (88.8 percent) components, respectively. The major components were aromadendrene (leaves 26.6 percent; stems 18.8 percent), sabinene (leaves 13.8 percent; stems 6.7 percent) and beta-caryophyllene (leaves 11.5 percent; stems 17.9 percent). A total of 60 (87.0 percent) and 39 (82.9 percent) components were identified from leaves and stems of P. ribesioides, respectively. The most abundant components were beta-caryophyllene (leaves 20.0 percent; stems 14.4 percent), camphene (leaves 16.3 percent; stems 12.3 percent) and delta-cadinene (leaves 4.4 percent; stems 7.8 percent). Antioxidant activity using DPPH and total phenolic content were tested for essential oils. However, the essential oils showed low antioxidant activity and phenolic content, compared to BHT. Studies of tyrosinase inhibition showed that the essential oils of P. ribesioides leaves had the highest inhibition (30.0 percent), although were lower than the control (kojic acid 81.8 percent). The evaluation of antimicrobial activities revealed that P. ribesioides essential oils showed strong activity against Bacillus cereus and Staphylococcus aureus, both with MIC value 62.5 micrograms/mL.

Se determinaron las composiciones químicas, las actividades antioxidante y antimicrobiana, y el contenido total de fenoles de los aceites esenciales de P. stylosum y P. ribesioides. El análisis GC y GC-MS de los aceites esenciaales de hojas y tallos de P. stylosum permitió la identificación de 50 (89.2 por ciento) y 45 (88.8 por ciento) de components, respectivamente. Los principales componentes fueron aromadendreno (hojas 26.6 por ciento; tallos 18.8 por ciento), sabineno (hojas 13.8 por ciento; tallos 6.7 por ciento) y beta-cariofileno (hoja 11.5 por ciento; tallo 17.9 por ciento). Se identificaron 60 (87.0 por ciento) y 39 (82.9 por ciento) components en los aceites esenciales de hojas y tallos de P. ribesioides. Los componentes más abundantes fueron beta-cariofileno (hojas 20.0 por ciento; tallos 14.4 por ciento), canfeno (hojas 16.3 por ciento; tallos 12.3 por ciento) y delta-cadineno (hojas 4.4 por ciento; tallos 7.8 por ciento). Los aceites esenciales se ensayaron para determinar sus actividades antioxidantes con DPPH y el contenido de fenoles totales. Para los aceites esenciales obtenidos se determinaron valores bajos en la actividad antioxidante con DPPH y el contenido total de fenoles, en comparación con BHT. Sin embargo, los ensayos de inhibición de tirosinasa most raron que el aceite esencial de las hojas de P. ribesioides presento la mayor inhibición (30.0 por ciento), aunque mas baja que el compuesto control (Àcido Kójico, 81.8 por ciento). Para el aceite esencial de P. ribesioides se determinó una MIC 62.5 mg/mL contra Bacillus cereus y Staphylococcus aureus.