ITS1 Barcode and Phytochemical Analysis by Gas Chromatography-Mass Spectrometry of Corynaea crassa Hook. f (Balanophoraceae) from Ecuador and Peru.

The use of medicinal plants is the basis of traditional healthcare. Recently, the use of herbal medicine has been increasing among consumers due to availability, economy, and less side effect. For instance, the hemiparasite plant Corynaea crassa has medicinal properties and could be found in some regions of America, from Costa Rica to Bolivia. Phytochemical and genetic characterization of medicinal plants is needed for proper identification of metabolites responsible for medicinal properties and for genotyping, respectively. Moreover, characterization of medicinal plants through the use of DNA barcodes is an important tool for phylogenetic analysis and identification of species; furthermore, complemented with phytochemical analysis, both are useful for identification of plant species and quality control of medicinal products. The objective of this study was to analyze the species of C. crassa collected in Ecuador and Peru from the phylogenetic and phytochemical point of view. Polymerase chain reaction (PCR) was performed for amplification of the internal transcribed spacer 1 (ITS1) region after DNA extraction of samples of C. crassa. Blast analysis was performed in the GenBank database with the ITS1 sequences obtained from two accessions of C. crassa from Ecuador (GenBank accession numbers OM471920 and OM471919 for isolates CIBE-17 and CIBE-18, respectively) and three from Peru (GenBank accession numbers OM471921, OM471922, and OM471923 for isolates CIBE-13, CIBE-14, and CIBE-15, respectively). The accessions available in the GenBank were used for phylogenetic analysis. For the phytochemical analysis, hydroalcoholic extracts were obtained by maceration using 80% ethanol as solvent, followed by a derivatization process and analysis by gas chromatography-mass spectrometry. Based on the phylogenetic analysis of the C. crassa samples, the ITS1 sequence could be used to differentiate C. crassa of different locations. The samples of C. crassa from Ecuador and Peru are more similar between them than with other clades including Helosis spp. The phytochemical study revealed differences in the presence and relative abundance of some metabolites; mainly eugenol, 1,4-lactone arabinonic acid, dimethoxyrabelomycin and azelaic acid, which are reported for the first time for the species under study and the genus Corynaea. These results are the first findings on the combined analysis using genetic and phytochemical analysis for C. crassa, which could be used as a useful tool for quality control of the C. crassa species in medicinal products.

Morphological and molecular barcode analysis of the medicinal tree Mimusops coriacea (A.DC.) Miq. collected in Ecuador.

BACKGROUND: Mimusops coriacea (A.DC.) Miq., (Sapotaceae), originated from Africa, were introduced to coastal areas in Ecuador where it is not extensively used as a traditional medicine to treat various human diseases. Different therapeutically uses of the species include: analgesic, antimicrobial, hypoglycemic, inflammation and pain relieve associated with bone and articulation-related diseases. Furthermore, Mimusops coriacea could be used as anti-oxidant agent. However, botanical, chemical or molecular barcode information related to this much used species is not available from Ecuador. In this study, morphological characterization was performed from leaves, stem and seeds. Furthermore, genetic characterization was performed using molecular barcodes for rbcL, matk, ITS1 and ITS2 using DNA extracted from leaves. METHODS: Macro-morphological description was performed on fresh leaves, stem and seeds. For anatomical evaluation, tissues were embedded in paraffin and transversal dissections were done following incubation with sodium hypochlorite and safranin for coloration and fixated later in glycerinated gelatin. DNA extraction was performed using a modified CTAB protocol from leaf tissues, while amplification by PCR was accomplished for the molecular barcodes rbcL, matK, ITS1 and ITS2. Sequence analysis was performed using blast in the GenBank. Phylogenetic analysis was performed with accessions queried in the GenBank belonging to the subfamily Sapotoideae. RESULTS: Leaf size was 13.56 ± 1.46 × 7.49 ± 0.65 cm; where is a macro-morphological description of the stem (see Methods). The peel of the seeds is dark brown. Sequence analysis revealed that amplicons were generated using the four barcodes selected. Phylogenetic analysis indicated that the barcodes rbcL and matK, were not discriminated between species within the same genus of the subfamily Sapotoideae. On the other hand, the ITS1 and ITS2 were discriminative at the level of genus and species of the Sapotoideae.

Comparative Pharmacognosy, Chemical Profile and Antioxidant Activity of Extracts from Phania matricarioides (Spreng.) Griseb. Collected from Different Localities in Cuba.

Phania matricarioides (Spreng.) Griseb. is a traditionally used plant with various pharmacological properties. However, there are only scarce reports about the phytochemistry and biological activity of this plant. In this work, P. matricarioides was collected from three different localities of Cuba: PmB (collected in Bauta, Artemisa), PmC (collected in Cangrejeras, Artemisa), and PmI (collected in La Lisa, Havana), extracted with aqueous ethanol, and analyzed macroscopically and microscopically. The extracts were screened for phytochemical contents, analyzed by TLC and HPLC, and screened for antioxidant activity using the FRAP and DPPH assays. Macroscopic analysis showed similar results for all samples; however, microscopic, physicochemical and phytochemical studies showed appreciable differences. In particular, the total solid of PmC extract was higher (1.94 ± 0.03%) than the other samples. In HPLC profiles, quercetin was identified in the three samples and a greater similarity between samples PmB and PmI was observed. All samples demonstrated radical-scavenging antioxidant activity by the DPPH assay, which PmC also demonstrated the smaller (p < 0.05) value (IC50 = 27.4 ± 0.1 µg/mL), but was statistically superior (p < 0.05) to vitamin C (IC50 = 23.7 ± 0 µg/mL). Also, in the FRAP assay, a higher vitamin C equivalent of PmC was significantly superior (p < 0.05) to the other extracts at the evaluated concentrations, which is likely due to a higher concentration of quercetin. In conclusion, P. matricarioides could constitute a potential resource in the field of phytotherapeutic products, and the results obtained can contribute to the development of the quality control norms for this species.

Efecto antiinflamatorio y antimicótico del extracto alcohólico y composición química del aceite de hojas de Conyza bonariensis (L) Cronquist (canilla de venado) / Anti-inflammatory and antimitotic effect of the alcoholic extract and chemical composition of the oil from Conyza bonariensis (L) Cronquist (deer shinbone) leaves

Introducción: la especie Conyza bonariensis (L.) Cronquist, comúnmente llamada canilla de venado, es de origen americano. Objetivos: contribuir al estudio farmacológico y toxicológico de los extractos alcohólicos y a la evaluación química del aceite de hojas de C bonariensis de Ecuador. Métodos: se emplearon 2 tinturas a 50 por ciento con etanol a 70 y 90 por ciento de hojas secas. Se realizó un estudio fitoquímico a través del sistema acoplado de cromatografía gaseosa-espectrometría de masas, se determinó el efecto antimicótico mediante inhibición de dilución en placa con siembra en superficie por diseminación, y antiinflamatorio en el modelo de edema de la oreja inducido por aceite de Croton en ratones albinos de la línea OF1. Resultados: el tamizaje fitoquímico mostró presencia de aceites esenciales, triterpenos-esteroides, compuestos fenólicos, flavonoides, quinonas, antocianidinas, saponinas y compuestos reductores. Se determinó estructuras a 32 componentes del aceite esencial extraído de las hojas y el sesquiterpeno: trans-b-farneseno resultó el componente mayoritario con 70,82 por ciento, este compuesto se informa por primera vez en esta especie. El extracto hidroalcohólico (25 µL a cada lado de las orejas tratadas) inhibió significativamente la inflamación comparado con bencidamina. Conclusiones: los extractos hidroalcohólicos mostraron efecto antiinflamatorio en las condiciones experimentales del estudio y se determinó la composición química del aceite de hojas de C. bonariensis(AU)

Introduction: the Conyza bonariensis (L) Cronquist species, usually called deer shinbone is of American origin. Objectives: to contribute to the pharmacological and toxicological study of alcoholic extracts and to the chemical assessment of C. bonariensis leaf oil from Ecuador. Methods: two 50 percent tinctures of 70 percent and 90 percent ethanol content in dry leaves were used. A phytochemical study was conducted by an combined system of mass spectrometry and gas chromatography; the amniotic effect was determined using plate dilution inhibition with surface culturing by dissemination, and the anti-inflammatory effect in the Croton oil induced ear edema model in OF1 line albino mice. Results: the phytochemical screening showed the presence of essential oils, triterpene-steroids, phenolic compounds, flavonoids, quinones, anthocyanidines, saponins and reducing compounds. The structures of 32 essential oil components extracted from the leaves were identified along with and the sesquiterpene trans-b-farnesene that was the major component (70.82 percent), this compound is reported for the first time in this species. The hydroalcoholic extract (25 µL on each side of the treated ears) significantly inhibited inflammation compared to benzidamine. Conclusions: hydroalcoholic extracts showed anti-inflammatory effect in the experimental conditions of the study and the chemical composition of C. bonariensis leaf oil was identified(AU)

Efecto antiinflamatorio y antimicótico del extracto alcohólico y composición química del aceite de hojas de Conyza bonariensis (L) Cronquist (canilla de venado) / Anti-inflammatory and antimitotic effect of the alcoholic extract and chemical composition of the oil from Conyza bonariensis (L) Cronquist (deer shinbone) leaves

Introducción: la especie Conyza bonariensis (L.) Cronquist, comúnmente llamada canilla de venado, es de origen americano. Objetivos: contribuir al estudio farmacológico y toxicológico de los extractos alcohólicos y a la evaluación química del aceite de hojas de C bonariensis de Ecuador. Métodos: se emplearon 2 tinturas a 50 por ciento con etanol a 70 y 90 por ciento de hojas secas. Se realizó un estudio fitoquímico a través del sistema acoplado de cromatografía gaseosa-espectrometría de masas, se determinó el efecto antimicótico mediante inhibición de dilución en placa con siembra en superficie por diseminación, y antiinflamatorio en el modelo de edema de la oreja inducido por aceite de Croton en ratones albinos de la línea OF1. Resultados: el tamizaje fitoquímico mostró presencia de aceites esenciales, triterpenos-esteroides, compuestos fenólicos, flavonoides, quinonas, antocianidinas, saponinas y compuestos reductores. Se determinó estructuras a 32 componentes del aceite esencial extraído de las hojas y el sesquiterpeno: trans-b-farneseno resultó el componente mayoritario con 70,82 por ciento, este compuesto se informa por primera vez en esta especie. El extracto hidroalcohólico (25 µL a cada lado de las orejas tratadas) inhibió significativamente la inflamación comparado con bencidamina. Conclusiones: los extractos hidroalcohólicos mostraron efecto antiinflamatorio en las condiciones experimentales del estudio y se determinó la composición química del aceite de hojas de C. bonariensis

Introduction: the Conyza bonariensis (L) Cronquist species, usually called deer shinbone is of American origin. Objectives: to contribute to the pharmacological and toxicological study of alcoholic extracts and to the chemical assessment of C. bonariensis leaf oil from Ecuador. Methods: two 50 percent tinctures of 70 percent and 90 percent ethanol content in dry leaves were used. A phytochemical study was conducted by an combined system of mass spectrometry and gas chromatography; the amniotic effect was determined using plate dilution inhibition with surface culturing by dissemination, and the anti-inflammatory effect in the Croton oil induced ear edema model in OF1 line albino mice. Results: the phytochemical screening showed the presence of essential oils, triterpene-steroids, phenolic compounds, flavonoids, quinones, anthocyanidines, saponins and reducing compounds. The structures of 32 essential oil components extracted from the leaves were identified along with and the sesquiterpene trans-b-farnesene that was the major component (70.82 percent), this compound is reported for the first time in this species. The hydroalcoholic extract (25 µL on each side of the treated ears) significantly inhibited inflammation compared to benzidamine. Conclusions: hydroalcoholic extracts showed anti-inflammatory effect in the experimental conditions of the study and the chemical composition of C. bonariensis leaf oil was identified

Efecto antiinflamatorio y composición química del aceite de ramas de Bursera graveolens Triana & Planch. (palo santo) de Ecuador / Antinflammatory effect and chemical composition of Bursera graveolens Triana & Planch. branch oil (palo santo) from Ecuador

INTRODUCCIÓN: la especie Bursera graveolens Triana & Planch, comúnmente llamada palo santo, es una planta nativa de las costas ecuatorianas y peruanas. OBJETIVOS: contribuir al estudio fitoquímico y farmacológico de la especie B. graveolens, nativa de San José de Ancón, provincia de Santa Elena, Ecuador. MÉTODOS: se empleó el extracto hidroalcohólico 50 porciento de ramas secas. Se realizó un estudio fitoquímico a través del sistema acoplado de cromatografía gaseosa-espectrometría de masas y se determinó el efecto antiinflamatorio en el modelo de edema de la oreja inducido por aceite de Crotón en ratones albinos de la línea OF1. RESULTADOS: el tamizaje fitoquímico mostró presencia de aceites esenciales, triterpenos-esteroides, compuestos fenólicos, flavonoides, quinonas, antocianidinas, saponinas y compuestos reductores. Se determinó estructuras a 11 componentes del aceite esencial extraído de las ramas y el sesquiterpeno denominado viridiflorol resultó el componente mayoritario con 70,82 porciento, este compuesto se informa por primera vez en esta especie. El extracto hidroalcohólico (25 mL a cada lado de las orejas tratadas) inhibió significativamente la inflamación comparado con la bencidamina. CONCLUSIONES: los extractos hidroalcohólicos mostraron un efecto antiinflamatorio en las condiciones experimentales del estudio y se determinó la composición química del aceite de ramas de B. graveolens de Ecuador(AU)

INTRODUCTION: Bursera graveolens Triana & Planch. species, commonly called palo santo, is an indigenous plant from Ecuadorian and Peruvian coasts. OBJECTIVES: to contribute to the phytochemical and pharmacological study of B. graveolens species from San José de Ancon, Santa Elena province, Ecuador. METHODS: 50 percent hydroalcoholic extract from dry branches was used. The phytochemical study was based on a combined system of gas chromatography-mass spectrometry and the antinflammatory effect was determined in the model of ear edema induced by Croton oil in albino OF1 mice. RESULTS: phytochemical screening showed the existence of essential oils, triterpen-steroids, phenolic compounds, flavonoids, quinones, antocyanidines, saponins and reducing compounds. The structures of eleven essential oil components from branches were determined whereas sesquiterpene called viridiflorol turned out to be the main component with 70,82 percent; this is the first time that this compound is reported in this species. The hydroalcoholic extract (25 mL applied at each side of the treated ears) significantly inhibited the inflammation compared with the effect of benzydamine. CONCLUSIONS: hydroalcoholic extracts showed antinflammatory effect under experimental study conditions and the chemical composition of branch oil from B. graveolens from Ecuador was determined(AU)

Efecto antiinflamatorio y composición química del aceite de ramas de bursera graveolens triana & planch. (palo santo) de Ecuador / Antinflammatory effect and chemical composition of bursera graveolens triana & planch. branch oil (palo santo) from Ecuador

INTRODUCCIÓN: la especie Bursera graveolens Triana & Planch, comúnmente llamada palo santo, es una planta nativa de las costas ecuatorianas y peruanas. OBJETIVOS: contribuir al estudio fitoquímico y farmacológico de la especie B. graveolens, nativa de San José de Ancón, provincia de Santa Elena, Ecuador. MÉTODOS: se empleó el extracto hidroalcohólico 50 porciento de ramas secas. Se realizó un estudio fitoquímico a través del sistema acoplado de cromatografía gaseosa-espectrometría de masas y se determinó el efecto antiinflamatorio en el modelo de edema de la oreja inducido por aceite de Crotón en ratones albinos de la línea OF1. RESULTADOS: el tamizaje fitoquímico mostró presencia de aceites esenciales, triterpenos-esteroides, compuestos fenólicos, flavonoides, quinonas, antocianidinas, saponinas y compuestos reductores. Se determinó estructuras a 11 componentes del aceite esencial extraído de las ramas y el sesquiterpeno denominado viridiflorol resultó el componente mayoritario con 70,82 porciento, este compuesto se informa por primera vez en esta especie. El extracto hidroalcohólico (25 mL a cada lado de las orejas tratadas) inhibió significativamente la inflamación comparado con la bencidamina. CONCLUSIONES: los extractos hidroalcohólicos mostraron un efecto antiinflamatorio en las condiciones experimentales del estudio y se determinó la composición química del aceite de ramas de B. graveolens de Ecuador.

INTRODUCTION: Bursera graveolens Triana & Planch. species, commonly called palo santo, is an indigenous plant from Ecuadorian and Peruvian coasts. OBJECTIVES: to contribute to the phytochemical and pharmacological study of B. graveolens species from San José de Ancon, Santa Elena province, Ecuador. METHODS: 50 percent hydroalcoholic extract from dry branches was used. The phytochemical study was based on a combined system of gas chromatography-mass spectrometry and the antinflammatory effect was determined in the model of ear edema induced by Croton oil in albino OF1 mice. RESULTS: phytochemical screening showed the existence of essential oils, triterpen-steroids, phenolic compounds, flavonoids, quinones, antocyanidines, saponins and reducing compounds. The structures of eleven essential oil components from branches were determined whereas sesquiterpene called viridiflorol turned out to be the main component with 70,82 percent; this is the first time that this compound is reported in this species. The hydroalcoholic extract (25 mL applied at each side of the treated ears) significantly inhibited the inflammation compared with the effect of benzydamine. CONCLUSIONS: hydroalcoholic extracts showed antinflammatory effect under experimental study conditions and the chemical composition of branch oil from B. graveolens from Ecuador was determined.

Studies on the toxicity of Punica granatum L. (Punicaceae) whole fruit extracts.

Current investigation focuses on the toxicity evaluation of whole fruit hydroalcoholic extract of Punica granatum L. (Punicaceae), used in Cuban traditional medicine a.o. for the treatment of respiratory diseases. Previous findings on the anti-influenza activity of Punica granatum extracts has given support to the ethnopharmacological application. In our study, in chick embryo model, it was found that doses of the extract of less than 0.1 mg per embryo are not toxic. The LD50 of the extract, determined in OF-1 mice of both sexes after intraperitoneal administration, was 731 mg/kg. Confidence limits were 565-945 mg/kg. At the doses of 0.4 and 1.2 mg/kg of extract, the repeated intranasal administration to Wistar rats produced no toxic effects in terms of food intake, weight gain, behavioural or biochemical parameters, or results of histopathological studies. We conclude that toxic effects of Punica granatum fruit extract occurred at higher doses than those effective in the models where the anti-viral activity has been studied or than those doses used in Cuban folk medicine.