Preformulation studies for the development of a microemulsion formulation from Ambrosia peruviana All, with anti-inflammatory effect

Abstract Natural products are considered an important source of the therapeutic arsenal currently available. Among these alternatives are the seeds of Ambrosia peruviana (altamisa), whose extract has shown an anti-inflammatory effect. The main objective of this work was to perform a preformulation study of Ambrosia peruviana seeds ethanolic extract, where the main factors that affect the physical, chemical, and pharmacological stability of the extract were evaluated, as well as a compatibility study by differential scanning calorimetry (DSC) analysis against different excipients. A dry extract was obtained by rotary evaporation of the seeds macerated with 96% ethanol. The anti-inflammatory activity was determined by measuring its effect on NO production in RAW 264.7 macrophages, stimulated with LPS. The results showed that the dry extract maintained its stability over time when stored at a temperature of 4 and 25ºC, demonstrating its biological activity, the content of phenolic compounds, and its physicochemical parameters remain practically invariable. However, when exposed to high temperatures (60 ºC) it was affected. The thermal analysis revelated that the behavior of most of the selected excipients and the dry extract was maintained, which indicates that it did not present incompatibilities, therefore they can be candidates for formulating a microemulsion.

Acaricidal Efficacy of Plants from Ecuador, Ambrosia peruviana (Asteraceae) and Lepechinia mutica (Lamiaceae) against Larvae and Engorged Adult Females of the Common Cattle Tick, Rhipicephalus microplus.

Control measures against common cattle tick Rhipicephalus microplus are of the upmost importance because of considerable, deleterious impact on a farm's economy. Due to resistance phenomena to synthetic acaricides being a constraint in affected farms, the search for plant derivatives as acaricides has increased dramatically in recent years. In this work, essential oils obtained from two Ecuadorian plants, Ambrosia peruviana and Lepechinia mutica (EOAp, EOLm), traditionally used as insecticides in indigenous communities, were studied on larvae and engorged females at the parasitic stages of R. microplus. Larvae and females were treated with five (0.0625, 0.125, 0.25, 0.50 and 1%) and six concentrations (0.125, 0.25, 0.50, 1, 2 and 4%), respectively, of each EOsAp/Lm. A 98-99% larval mortality was achieved with 0.5% of both EOsAp/Lm. EOAp inhibited oviposition and egg hatching up to 82% and 80%, respectively, and had an overall efficacy of 93.12%. Efficacy of EOLm was 72.84%, due to the low influence of EOLm on reproductive parameters. By steam distillation and GC-MS analysis, γ-Curcumene was identified as the main constituent (52.02%) in the EOAp and Shyobunol (10.80%) in EOLm. The results suggest that major components of both essential oils should be further studied as promissory acaricides against R. microplus.

Actividad antibacterial y larvicida sobre Aedes aegypty L. de extractos de Ambrosia peruviana Willd (Altamisa) / Antibacterial and larvicidal activity against Aedes aegypti L. of extracts from Ambrosia peruviana Willd (altamisa)

Introducción: en la actualidad, nuevas tendencias tecnológicas e iniciativas se están presentando en el desarrollo de productos insecticidas derivados de productos naturales, y de nuevos agentes antimicrobianos, dado que poseen bioactivos que son selectivos, biodegradables y tienen menores efectos adversos. La especie Ambrosia peruviana Willd. es de gran interés en el estudio por su gran potencial biológico y etnobotánico. Objetivo: evaluar la actividad larvicida sobre Aedes aegypty L. y la actividad antibacteriana sobre bacterias Gram positivas y Gram negativas de extractos de A. peruviana. Métodos: a partir del material vegetal seco (hojas), se obtuvieron cinco extractos de diferente polaridad en hexano (H), diclorometano (D), acetato de etilo (A) y etanol (E) y aceites esenciales (AE), los cuales fueron evaluados mediante la inhibición del crecimiento de larvas por el método recomendado de la OMS y la inhibición de las bacterias por el método de difusión en agar de Kirby-Bauer. Resultados: la tasa de mortalidad encontrada a las 24 h a una concentración de 200 ppm para todos los extractos fue del 10 por ciento. Al evaluar el paso de los insectos de larvas a adultos a las 144 h se observó a esta misma concentración una mortalidad del 100 por ciento con todos los extractos. Por otra parte, los extractos de A. peruviana presentaron inhibición sobre Bacillus cereus Frankland & Frankland y Bacillus subtilis (Ehrenberg) Cohn con halos de inhibición del extracto de diclorometano (APExtD) de 10,5 y 15,0 mm de diámetro respectivamente, al contrario sobre las cepas Serratia marcescens Bizio, Proteus mirabilis Hauser, Enterobacter cloacae (Jordan) Hormaeche & Edwards y Staphylococcus aureus Rosenbach no se presentó actividad antibacteriana. Conclusiones: esta investigación es el primer reporte de actividad larvicida sobre A. aegypty y de actividad antibacteriana sobre B. cereus y B. subtilis de varios extractos de A. peruviana con promisorios resultados en estos modelos(AU)

Introduction: New technological trends and initiatives are currently being put forth concerning the development of insecticidal products and antimicrobial agents of natural origin, since their bioactive components are selective and biodegradable, and cause fewer adverse effects. The species Ambrosia peruviana Willd. was of great interest to the present study, due to its great biological and ethnobotanical potential. Objective: Evaluate the larvicidal activity of A. peruviana extracts against Aedes aegypti L., and its antibacterial activity against gram-positive and gram-negative bacteria. Methods: Dry plant material (leaves) was processed to obtain five extracts of different polarity in hexane (H), dichloromethane (D), ethyl acetate (A), ethanol (E) and essential oils (AE), which were evaluated for larval growth inhibition with the method recommended by WHO, and for bacterial inhibition with the Kirby-Bauer agar diffusion method. Results: The mortality rate at 24 h and a concentration of 200 ppm was 10 percent for all extracts. Examination of the transition of larvae into adults at 144 h and the same concentration revealed a mortality of 100 percent with all extracts. On the other hand, the extracts of A. peruviana displayed inhibition capacity against Bacillus cereus Frankland & Frankland and Bacillus subtilis (Ehrenberg) Cohn with inhibition haloes for the dichloromethane extract (APExtD) of 10.5 and 15.0 mm in diameter, respectively, whereas no antibacterial activity was found against the strains Serratia marcescens Bizio, Proteus mirabilis Hauser, Enterobacter cloacae (Jordan) Hormaeche & Edwards and Staphylococcus aureus. Conclusions: This study is the first report of larvicidal activity againstA. aegypti and antibacterial activity against B. cereus and B. subtilis by several extracts of A. peruviana with promising results in these models(AU)

Actividad antibacterial y larvicida sobre Aedes aegypty L. de extractos de Ambrosia peruviana Willd (Altamisa) / Antibacterial and larvicidal activity against Aedes aegypti L. of extracts from Ambrosia peruviana Willd (altamisa)

Introducción: en la actualidad, nuevas tendencias tecnológicas e iniciativas se están presentando en el desarrollo de productos insecticidas derivados de productos naturales, y de nuevos agentes antimicrobianos, dado que poseen bioactivos que son selectivos, biodegradables y tienen menores efectos adversos. La especie Ambrosia peruviana Willd. es de gran interés en el estudio por su gran potencial biológico y etnobotánico. Objetivo: evaluar la actividad larvicida sobre Aedes aegypty L. y la actividad antibacteriana sobre bacterias Gram positivas y Gram negativas de extractos de A. peruviana. Métodos: a partir del material vegetal seco (hojas), se obtuvieron cinco extractos de diferente polaridad en hexano (H), diclorometano (D), acetato de etilo (A) y etanol (E) y aceites esenciales (AE), los cuales fueron evaluados mediante la inhibición del crecimiento de larvas por el método recomendado de la OMS y la inhibición de las bacterias por el método de difusión en agar de Kirby-Bauer. Resultados: la tasa de mortalidad encontrada a las 24 h a una concentración de 200 ppm para todos los extractos fue del 10 por ciento. Al evaluar el paso de los insectos de larvas a adultos a las 144 h se observó a esta misma concentración una mortalidad del 100 por ciento con todos los extractos. Por otra parte, los extractos de A. peruviana presentaron inhibición sobre Bacillus cereus Frankland & Frankland y Bacillus subtilis (Ehrenberg) Cohn con halos de inhibición del extracto de diclorometano (APExtD) de 10,5 y 15,0 mm de diámetro respectivamente, al contrario sobre las cepas Serratia marcescens Bizio, Proteus mirabilis Hauser, Enterobacter cloacae (Jordan) Hormaeche & Edwards y Staphylococcus aureus Rosenbach no se presentó actividad antibacteriana. Conclusiones: esta investigación es el primer reporte de actividad larvicida sobre A. aegypty y de actividad antibacteriana sobre B. cereus y B. subtilis de varios extractos de A. peruviana con promisorios resultados en estos modelos(AU)

Introduction: New technological trends and initiatives are currently being put forth concerning the development of insecticidal products and antimicrobial agents of natural origin, since their bioactive components are selective and biodegradable, and cause fewer adverse effects. The species Ambrosia peruviana Willd. was of great interest to the present study, due to its great biological and ethnobotanical potential. Objective: Evaluate the larvicidal activity of A. peruviana extracts against Aedes aegypti L., and its antibacterial activity against gram-positive and gram-negative bacteria. Methods: Dry plant material (leaves) was processed to obtain five extracts of different polarity in hexane (H), dichloromethane (D), ethyl acetate (A), ethanol (E) and essential oils (AE), which were evaluated for larval growth inhibition with the method recommended by WHO, and for bacterial inhibition with the Kirby-Bauer agar diffusion method. Results: The mortality rate at 24 h and a concentration of 200 ppm was 10 percent for all extracts. Examination of the transition of larvae into adults at 144 h and the same concentration revealed a mortality of 100 percent with all extracts. On the other hand, the extracts of A. peruviana displayed inhibition capacity against Bacillus cereus Frankland & Frankland and Bacillus subtilis (Ehrenberg) Cohn with inhibition haloes for the dichloromethane extract (APExtD) of 10.5 and 15.0 mm in diameter, respectively, whereas no antibacterial activity was found against the strains Serratia marcescens Bizio, Proteus mirabilis Hauser, Enterobacter cloacae (Jordan) Hormaeche & Edwards and Staphylococcus aureus. Conclusions: This study is the first report of larvicidal activity againstA. aegypti and antibacterial activity against B. cereus and B. subtilis by several extracts of A. peruviana with promising results in these models(AU)

Detección de metabolitos secundarios en Ambrosia peruviana Willd y determinación de la actividad antibacteriana y antihelmíntica / Secondary metabolite detection in Ambrosia peruviana Willd and determination of antibacterial and antihelmintic activity

En el conocimiento etnofarmacológico, Ambrosia peruviana es conocida como una planta antiinfecciosa y antihelmíntica. Dados los altos índices de resistencia bacteriana y parasitaria, se realizó la tamización fitoquímica preliminar siguiendo la metodología de Sanabria, y ensayos biológicos frente a aislamientos clínicos bacterianos, parásitos caninos y Artemia salina. Los ensayos de citotoxicidad en A. salina se realizaron por exposición de los adultos a concentraciones variables de los extractos. La actividad antibacteriana se realizó por los métodos de difusión en disco y concentración inhibitoria mínima (CIM). Se determinó el porcentaje de huevos que tenían el embrión del parásito, en el medio de cultivo se mantuvieron en lactato de Ringer con suplemento al 10% de RPMI y 1X ATM. Se determinó el porcentaje de huevos con embrión, o fecundados, liberados en el medio de cultivo con adición de extracto etanólico y acuoso secos de A. peruviana. Los ensayos de la especie vegetal frente a helmintos se realizaron al sumergir los adultos en medio con suplemento de extracto etanólico seco (usando diferentes concentraciones) y con fracciones ricas en alcaloides. Se identificó la presencia de alcaloides, glucósidos cardiotónicos, quinonas, flavonoides, carbohidratos, taninos y saponinas. La concentración letal media para el extracto etanólico seco fue 64,2 µg/ml, mientras que para el extracto acuoso fue de 840,4 µg/ml. Los extractos no presentaron actividad antibacteriana. Los ejemplares adultos de T. canis presentaron disminución de la motilidad frente a extractos secos, mientras que en la fracción de alcaloides murieron luego de 4 horas de exposición. Los extractos de A. peruviana sobre huevos de T. canis permitieron una disminución en el porcentaje de huevos con embrión que no dependía del extracto observado sino de la concentración empleada.

Ambrosia peruviana has been reported as an anti-infective and anti-parasitic plant in the ethno-pharmacological environment. Given the high rates of bacterial and parasitic resistance against commercial drugs recorded, we performed the preliminary phytochemical screening following Sanabria´s method and biological tests against clinical bacterial isolates, dog parasites, and Artemia salina. Cytotoxicity tests in Artemia salina were carried out by exposing adults to extracts of varied concentrations. The antibacterial activity was performed using the disk diffusion method and CIM. Toxocara canis nematodes were kept in Ringer´s lactate supplemented with 10% RPMI and 1X ATM. The embryo generation percentage of eggs released into the culture medium was evaluated adding ethanol and aqueous extracts of dried A. peruviana. The tests of A. peruviana against helminths were performed by immersing adults in a medium supplemented with dried ethanol extract (at various concentrations) and fractions rich in alkaloids. Phytochemical screening allowed preliminary identification of alkaloids, cardiotonic glucosides, quinones, flavonoids, carbohydrates, tannins and saponins. The LC50 for dry ethanol extract was 64.2µg/ml, while for the aqueous extract was 840.4µg/ml. The extracts did not show antibacterial activity. T. canis adults showed motility decrease against dried extracts, not so in alkaloid fraction where they died after a 4h exposure. The use of extracts from A. peruviana over Toxocara canis eggs caused a decrease in the percentage of embryo generation, which did not depend on the extract observed but on the concentration used.