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.

Study of Antipyretic Activity of Parthenium Hysterophorus in Rats

Introduction: Parthenium hysterophorus is an aggressive ubiquitous annual herbaceous weed with no economic importance unravelled till now. The decoction of P. Hysterophorus has been used in traditional medicine to treat fever, diarrhoea, neurologic disorders, urinary tract infections, dysentery, malaria and as emmenagogue. Objective: The study was conducted to screen the antipyretic activity of hydro-alcoholic extracts of Parthenium Hysterophorus. Materials and Methods: Thirty healthy rats weighing between 200-250gms were divided into five groups of six animals each, with 50% sex ratio. The initial rectal temperature of each animal was recorded by digital thermometer. The pyrexia was induced by injecting 20ml/kg (s.c.) of 20 per cent aqueous suspension of Brewer’s yeast in normal saline below the nape of the neck and rectal temperature was recorded by clinical thermometer immediately before (18 h) and 18 h after (0 h) Brewer’s yeast injection .The difference in temperature between 0 hour and respective time interval was found out by statistical method. The potency of extract to bring down the temperature was compared with that of the control group. Results: PH extract at 200 and 400 mg/kg produced significant antipyretic activity at 1 h after drug administration, whereas PH extract (800 mg/kg) and aspirin(300 mg/kg) showed significant antipyretic activity throughout the observation period up to 6 h. The extract showed marked antipyretic activity in a dose dependent manner. Conclusion: Parthenium hysterophorus has marked antipyretic activity.

Parthenium dermatitis in India: Past, present and future.

Parthenium dermatitis is an immuno-inflammatory disease caused by Parthenium hysterophorus and is the commonest cause of plant dermatitis in India. It is caused by airborne dry and friable plant particles including trichomes, and the most important allergens responsible for allergic contact dermatitis are sesquiterpene lactones. The combined type IV and type I hypersensitivity to parthenium has been recently postulated. In sensitized individuals, it can cause a spectrum of clinical patterns, such as classical airborne pattern, chronic actinic dermatitis-like presentation, mixed pattern dermatitis, exfoliative dermatitis, widespread dermatitis, and other rare patterns. There is definite trend towards change from airborne pattern to chronic actinic pattern in natural history of parthenium dermatitis. Contact sensitivity to parthenium is everlasting, and hence the disease runs a chronic course with exacerbation during summers. Patch testing with acetone or aqueous plant extract is the simplest way of confirming parthenium contact allergy. Management includes avoiding contact with allergen, managing dermatitis with topical corticosteroids/tacrolimus, and other immunosupressives like azathioprine. In future, we expect parthenium dermatitis to become less prevalent due to rapid urbanization and possible development of new biological methods to eradicate the parthenium. Genetic factors associated with susceptibility to parthenium dermatitis need to be studied.

Production and extraction of phytotoxins from Colletotrichum dematium FGCC# 20 effective against Parthenium hysterophorus L

The aim of this work was to study the herbicidal potential of Cell free culture filtrate of Colletotrichum dematium FGCC#20 against Parthenium by employing different bioassays i.e. shoot-cut, seedling, detached leaf and seed germination. On solvent extraction of the Cell free culture filtrate, Ethyl acetate extracted fraction showed the presence of phytotoxic moiety.

Especificidad de la cromatografía líquida de alta eficacia para evaluar estabilidad química basada en partenina del follaje seco pulverizado de Parthenium hysterophorus L (escoba amarga) / Specificity of HPLC to assess the chemical stability based on partenine from Parthenium hysterophorus L powdered dry foliage (escoba amarga)

INTRODUCCIÓN: se requiere una técnica de análisis específica que permita dar seguimiento al estudio de la estabilidad intrínseca del follaje seco pulverizado de Parthenium hysterophorus L. (escoba amarga), para la obtención de una forma farmacéutica de utilidad antiparasitaria con los requisitos de calidad, seguridad y eficacia exigidos. OBJETIVO: demostrar la especificidad de la técnica analítica de cromatografía líquida de alta eficacia para la cuantificación de partenina en el polvo de P. hysterophorus para su aplicación en estudios de estabilidad. MÉTODOS: se aplicó cromatografía líquida de alta eficacia a muestras degradadas de P. hysterophorus, bajo condiciones degradativas en medio ácido, básico y oxidativo. Se evaluó la especificidad de la técnica de análisis para detectar el componente de interés sin interferencias de sus productos de degradación y su posible utilidad en estudios de estabilidad del polvo de la planta. RESULTADOS: la cromatografía líquida de alta eficacia para cuantificar partenina en el polvo de P. hysterophorus resultó específica en las condiciones de trabajo establecidas y puede emplearse en estudios de estabilidad del sólido en el polvo de la planta. CONCLUSIONES: la técnica de cromatografía líquida de alta eficacia propuesta es específica y se recomienda su utilización en los estudios de estabilidad del sólido en polvo de la planta.

INTRODUCTION: it is required a specific analysis technique allowing the follow-up to stability study intrinsic of Parthenium hysterophorus L. (escoba amarga) powdered dry foliage to achieve in a pharmaceutical way a antiparasitic usefulness with the quality, safety and effectiveness demanded requirements. OBJECTIVE: to demonstrate the specificity of analytical technique of high-performance liquid chromatography for quantization of partenine in the powder of P. hysterophorus for its application in stability studies. METHODS: high performance liquid chromatography was applied to P. hysterophorus degraded samples under degradation conditions in an oxidative, basic and acid medium.The analysis technique specificity was assessed to detect the interest component without interferences of its degradation products and its possible usefulness in studies on solid stability in the plant powder. RESULTS: high-performance liquid chromatography to quantify the presence of partenine in the P. hysterophorus powder was specific in established work conditions and may be used in solid stability studies of plant powder. CONCLUSIONS: the high-performance proposed is specific and it is recommended in solid stability studies of solids in plant powder.

Estabilidad del polvo de Parthenium hysterophorus L (escoba amarga) basado en el contenido de partenina mediante cromatografía líquida de alta eficacia / Parthenium hysterophorus L. (escoba amarga) powder stability on the basis of parthenin contents by using high-performance liquid chromatography

INTRODUCCIÓN: Parthenium hysterophorus L. (escoba amarga) es una planta de reconocida acción antiparasitaria, carece de investigaciones analíticas y tecnológicas que permitan dar seguimiento a la estabilidad intrínseca del sólido en polvo de la planta, para la futura obtención de una forma farmacéutica de utilidad antiparasitaria científicamente fundamentada. OBJETIVO: establecer la estabilidad integral del polvo obtenido de follaje de P. hysterophorus en condiciones reales de almacenamiento basado en partenina. MÉTODOS: se desarrolló un estudio de la estabilidad intrínseca del sólido en polvo de P. hysterophorus frente a factores externos (humedad, temperatura y luz). RESULTADOS: la cromatografía líquida de alta eficacia (CLAE) para la cuantificación de partenina en el sólido en polvo del P. hysterophorus es específica y puede emplearse en los estudios de estabilidad. Se demostró que la humedad y la luz son factores que influyen negativamente en la estabilidad química de la partenina. CONCLUSIONES: la estabilidad química del sólido en polvo, conservado en frascos de vidrio ámbar cubiertos con papel de aluminio y tapón de goma, evaluada según las normativas ICH (2005), es insuficiente para su comercialización como materia prima para la elaboración de formas de dosificación.

INTRODUCTION: Parthenium hysterophorus (escoba amarga) is a well-known antiparasitic plant, but the lack of analytical and technological research studies does not allow following-up the intrinsic stability of the plant solid powder for obtaining a scientifically-proved pharmaceutical form with antiparasitic action. OBJECTIVE: to ascertain the integral stability of the powder from P. hysterophorus foliage under actual storage conditions, taking the parthenin content as a basis. METHODS: the intrinsic P. hysterophorus solid powder stability was determined against external factors such as humidity, temperature and light. RESULTS: the high-performance liquid chromatography is a specific technique to quantify the parthenin contents in P. hysterophorus powder and may be used in the stability studies. It was proved that humidity and light had a negative impact on the chemical stability of the parthenin. CONCLUSIONS: the chemical stability of the solid powder, preserved in amber glass flasks covered with aluminium paper and rubber cap and evaluated according to ICH standards (2005), is not adequate for this substance to be marketed as raw material for dosage form preparations.

Bio-control potential of Cladosporium sp. (MCPL - 461), against a noxious weed Parthenium hysterophorus L.

The phenological survey of Parthenium hysterophorus L., in and around the campus of Banaras Hindu University (BHU) was done for about two years (2004-06). During Nov. 2004, a few Parthenium plants were found diseased, and symptoms were restricted to the flowers, buds, and inflorescences. The disease causes sterility and reduces seed viability, which was observed with seed germination test from infected and healthy plants. The fungal pathogen was isolated and identified as Cladosporium sp. (MCPL-461). The severity of pathogen to the reproductive organs led to serious damages of the Parthenium plants. Thus in vitro and in vivo experiments were conducted to determine the bio-control potential of Cladosporium sp. (MCPL 461) against Parthenium weed. A combinatorial effort of Cladosporium sp. (MCPL 461) bio-control potential was evaluated with different culture media, incubation periods and spores strength. Spore suspension of 105 to 1012 spores ml-1 were used to spray on healthy Parthenium plants, and it was found that severe infection symptoms were appeared at 1010 to 1012 spores ml-1 suspension. LD50 was found at 107spores ml-1. To enhance the myco-herbicide activity 3% sucrose was added to the spore suspension, which further resolute the bio-control efficacy of the isolates. Only 20-30 % seeds of infected plants could germinate. However the safety of non-targeted and wild plants was also tested with Lantana camera, Chromolaena odorata and found that suspension up to 1012 spores ml-1 were not sufficient for disease outbreak in them.