Comparative study of antioxidant properties of the extracts of Polygonum acre H.B.K

SUMMARY Aims: This study aimed to characterize and compare the antioxidative potential of extracts obtained by infusion, decoction, tincture, aqueous extraction, and hydro-ethanolic maceration from the aerial parts of Polygonum acre H.B.K., which has been traditionally used in herbal preparations, for different purposes. The therapeutic benefits are attributed to phenolic compounds and their antioxidant properties. Methods: All extracts were characterized considering their quantitative content of the total phenolics, flavonoids, condensed and, hydrolysable tannins, by colori-metric methods. EHW-PA was selected for HPLC analysis as it showed a higher yield (10.58 % w/m) and a phenolic content > 200 mg GAE/g dry extract. The phenolic profile showed a chromatogram with 20 peaks, and the presence of gallic acid, rutin, and quercetin was verified by comparison with the retention times of standard compounds. The antioxidant activities were determined by ABTS capture test, ferric reducing antioxidant power test (FRAP), and the superoxide anion scavenging test. Results: Tinctures showed a higher average content of phenolic compounds, present mainly as flavonoid content. A significant correlation coefficient was observed between the total phenolic content and its antioxidant activity, determining by ABTS and FRAP assays. Differently, a low to moderate correlation between the flavonoid content and antioxidant activity was verified. Conclusion: This study reinforces the ethnopharmacological relevance of the Polygonum genus and could contribute to the scientific basis for the use of P. acre preparations.

Objetivos: Este estudio tuvo como objetivo caracterizar y comparar el potencial antioxidante de extractos obtenidos por infusión, decocción, tintura, extracción acuosa e hidroetanólica de las partes aéreas de Polygonum acre H.B.K., que se ha utilizado tradicionalmente en preparaciones a base de hierbas, para diferentes fines. Los beneficios terapéuticos se atribuyen a los compuestos fenólicos y sus propiedades antioxidantes. Métodos: Todos los extractos se caracterizaron considerando su contenido cuantitativo de fenólicos totales, flavonoides, taninos condensados e hidrolizables, por métodos colorimétricos. Se seleccionó EHW-PA para el análisis de HPLC ya que mostró un rendimiento más alto (10,58% m/m) y un contenido fenólico > 200 mg GAE/g de extracto seco. El perfil fenólico mostró un cromatograma con 20 picos y se verificó la presencia de ácido gálico, rutina y quercetina por comparación con los tiempos de retención de los compuestos estándar. Las actividades antioxidantes se determinaron mediante la prueba de captura ABTS, la prueba del poder antioxidante reductor férrico (FRAP) y la prueba de eliminación del anión superóxido. Resultados: Las tinturas mostraron un mayor contenido promedio de compuestos fenólicos, presentes principalmente como contenido de flavonoides. Se observó un coeficiente de correlación significativo entre el contenido fenólico total y su actividad antioxidante, determinado por ensayos ABTS y FRAP. De manera diferente, se verificó una correlación de baja a moderada entre el contenido de flavonoides y la actividad antioxidante. Conclusión: Este estudio refuerza la relevancia etnofarma-cológica del género Polygonum y podría contribuir a la base científica para el uso de preparaciones de P. acre.

Objetivos: O presente estudo teve como objetivo caracterizar e comparar o potencial antioxidante de extratos obtidos por infusão, decocção, tintura, extração aquosa e maceração hidroetanólica da parte aérea de Polygonum acre H.B.K., tradicional- mente utilizado em preparações fitoterápicas, para diversos fins. Os benefícios terapêuticos são atribuídos aos compostos fenólicos e suas propriedades antioxidantes. Métodos: Todos os extratos foram caracterizados quanto ao teor quantitativo de fenólicos totais, flavonoides, taninos condensados e hidrolisáveis, por métodos colorimétricos. O EHW-PA foi selecionado para análise por HPLC por apresentar maior rendimento (10,58% m/v) e conteúdo fenólico > 200 mg GAE/g de extrato seco. O perfil fenólico apresentou cromatograma com 20 picos, e a presença de ácido gálico, rutina e quercetina foi verificada pela comparação com os tempos de retenção dos compostos padrão. As atividades antioxidantes foram determinadas pelo teste de captura do radical ABTS, ensaio do potencial antioxidante por redução férrica (FRAP) e teste de desativação do ânion superóxido. Resultados: As tinturas apresentaram maior teor médio de compostos fenólicos, presentes, principalmente, na forma de flavonoides. Foi observado um coeficiente de correlação significativo entre o conteúdo fenólico total e sua atividade antioxidante, determinado pelos ensaios de ABTS e FRAP. Diferentemente, verificou-se uma correlação baixa a moderada entre o conteúdo de flavonoides e a atividade antioxidante. Conclusão: Este estudo reforça a relevância etnofarmacológica do gênero Polygonum e pode contribuir para a fundamentação científica do uso de preparações de P. acre.

Chemical Characterization of Opuntia ficus-indica (L.) Mill. Hydroalcoholic Extract and Its Efficiency against Gastrointestinal Nematodes of Sheep.

Opuntia ficus-indica (L.) Mill. is a xerophylous plant that originated in tropical and subtropical America. This plant is popularly known in Brazil as "palma forrageira" (cactus pear) and plays a fundamental role in animal nutrition, mainly in the Northeastern semi-arid region of the country. The plant has several uses since it presents bioactive compounds that confer biological and pharmacological properties. In this context, the cactus pear can also be considered a potential product to combat parasite infections. The objective of this study was to chemically characterize the O. ficus-indica hydroalcoholic extract (OFIEOH) and to determine its efficacy against gastrointestinal parasites using in vitro tests. Initially, the hydroalcoholic extract from cladode peels of O. ficus-indica was produced by maceration for 21 days. For the chemical characterization, colorimetric dosages were performed for carbohydrates, proteins, phenols and condensed tannins. Liquid chromatography coupled to mass spectrometry/electron spray ionization (LC-MS/ESI) was used to characterize the polyphenolic profile of the OFIEOH extract. Fifteen compounds were identified in the OFIEOH extract, such as methyl, glycosylated and aglycone quercetin derivatives and aglycone and glycosylated kaempferol derivatives. Tri-glycosylated methyl quercetin derivatives were the main compounds identified. In vitro egg hatch (EHT) and larval migration tests (LMT) were used in a range of concentrations of OFIEOH from 12.5 to 100 mg/mL for EHT and 12.5 to 200 mg/mL for LMT. In addition, the LMT was used to test ivermectin (IVM) (from 11.4 to 57.1 µM), associated with the inhibitory concentration of 50% (IC50) for OFIEOH. The combination of OFIEOH (12.5 to 200 mg/mL) plus the IC50 of IVM was also tested. The efficacy of OFIEOH alone varied from 19.33 to 90.0% using the EHT. The LMT revealed an efficacy of 5.78 to 77.26% for the extract. Both tests showed a concentration-dependence inhibitory effect. We found a drug-extract antagonistic neutralizing effect when doses of IVM were added to OFIEOH (maximum efficacy of 73.78%), while a positive additive effect was observed when OFIEOH was added to the IC50 of IVM (IC50 of 82.79 for OFIEOH alone against an IC50 of 55.08 of OFIEOH + IVM). The data from this work indicate that OFIEOH alone may be considered as a suitable ecofriendly product to control gastrointestinal parasites of sheep, offering a more holistic approach to improve animal farming and welfare. The drug-extract interaction is also a promising therapeutic alternative, reducing the final dose to the host, with an optimum combination effect.

Assessment of anthelmintic activity and bio-guided chemical analysis of Persea americana seed extracts.

The aim of this study was to characterize the extracts and fractions of Persea americana Mill. (Avocado) seeds and to determine the composition and the in vitro anthelmintic activity against third-stage larvae (L3) of Haemonchus contortus. The fresh (F) and dried (H) avocado seeds (PA) were subjected to extraction with hot water (W-PAF, W-PAH), ethanol (E-PAF, E-PAH) or methanol 70% (v/v), and partition with solvents of increasing polarity [n-hexane (H-PAF, H-PAH), chloroform (C-PAF, C-PAH), ethyl acetate (Ea-PAF, Ea-PAH), and n-butanol (B-PAF, B-PAH)], yielding a total of 14 extracts/fractions. After considering the yield, water solubility, and the preliminary results of the larval migration test (LMT), the E-PAF, E-PAH, H-PAF, and H-PAH were selected for further experiments. E-PAH presented an efficiency concentration of 50% (EC50) of 36 µg/mL on the LMT. E-PAH showed the greatest efficiency when its EC50 was compared to the other fractions (E-PAF = 147 µg/mL; H-PAF = 801 µg/mL; H-PAH = 77 µg/mL). After that, the E-PAH was chemically characterized, considering its quantitative polyphenolic and flavonoid contents by colorimetric and chromatographic techniques. E-PAH presented 50, 38, and 24 mg/g of dry matter of total phenol, condensed tannins (CT), and flavonoid contents, respectively. Using high performance liquid chromatography (HPLC) analysis, E-PAH had shown to have epicatechin (4.7 µg/mL), rutin (2.8 µg/mL), and chlorogenic acid (1.4 µg/mL) as its main constituents besides quercetin. These isolated compounds were evaluated using the LMT in order to relate the composition to the anthelmintic activity observed for E-PAH. Quercetin (EC50 = 7.8 µg/mL) and epicatechin (EC50 = 10 µg/mL) presented a higher efficiency than rutin (EC50 = 30 µg/mL). Chlorogenic acid was also tested with the LMT but did not present a significant efficiency. According to the results, the phenolic composition of E-PAH and the EC50 values obtained for the isolated phenols, it can be suggested that, besides the CT content, the presence of epicatechin and rutin contributed to the larvicidal activity of E-PAH. In conclusion, avocado seeds may be used as a source of polyphenols with promising anthelmintic applications.

Arabinogalactan-proteins from cell suspension cultures of Araucaria angustifolia.

Arabinogalactan-proteins (AGPs), found in the culture medium of suspension cells of Araucaria angustifolia grown in plant growth regulator-free and plant growth regulator-containing BM media, BM0 and BM2, respectively, were evaluated quantitatively and qualitatively. The concentrated extracellular fractions (CEFs), obtained from suspension cell cultures grown for 20 days in BM0 and BM2 media yielded two fractions, CEF-0 and CEF-2, respectively. CEF-0 and CEF-2 was submitted to selective precipitation using the beta-glucosyl Yariv reagent (beta-GlcY) to isolate AGPs for structural characterization; this yielded fractions designated CEF-0YPF and CEF-2YPF, respectively. The monosaccharide composition analysis established that samples were composed of Rha, Ara, Gal and uronic acid in a molar ratio 3:37:55:5 (CEF-0YPF) and 1:37:58:4 (CEF-2YPF), although trace amounts (<0.5 mol%) of Xyl were also found. Methylation analysis of CEF-YPF fractions showed similar results for both CEF-0YPF and CEF-2YPF, with non-reducing terminal units of Araf, Arap, Galp, Rhap and Xylp, as well as 3-O-substituted and 5-O-substituted Araf units and 3-O-substituted, 6-O-substituted and 3,6-di-O-substituted Galp units. The amino acid composition analysis established Ser, Ala, and Hyp as major amino acids in both samples. In conclusion, this investigation has shown that CEF-0YPF and CEF-2YPF contain macromolecules having typical AGP characteristics, including a Hyp/Ala/Ser-rich protein moiety, a (1-->3) and/or (1-->6) linked beta-d-galactopyranosyl main chain substituted by Gal, Ara, Rha and Xyl residues, and binding affinity for beta-GlcY and monoclonal anti-AGP antibodies.