RESUMO
It was shown that different extracts had significant differences in the toxicity of Polygonum multiflorum. In this study, the effect of sample preparation on components and liver toxicity of different extracts from P. multiflorum were determined. Hepatoxic components were discovered based on biomembrane extraction. Comparative chemistry and toxicology between ethanol and water extracts were also performed. The results showed that ethanol extract had much stronger hepatotoxicity, the content of emodin-8-O-ß-d-glucopyranoside, physcion-8-O-ß-d-glucopyranoside, emodin and physcion was significantly higher in ethanol extract than in water extract, while the human hepatocytes extraction showed that 2,3,5,4'-tetrahydroxystilbene-2-O-ß-d-glucopyranoside, emodin-8-O-ß-d-glucopyranoside, physcion-8-O-ß-d-glucopyranoside, emodin and physcion had interaction with human hepatocytes. The hepatotoxic effect of these components was investigated on human hepatocytes LO2 cells and emodin-8-O-ß-d-glucopyranoside, physcion-8-O-ß-d-glucopyranoside, emodin and physcion were finally confirmed to be, at least partial, hepatotoxic components. The results showed that sample preparation has significant effect on components in extracts of P. multiflorum especially the components related to hepatotoxicity. Water extract, the conventional administration form of Chinese herbs, is prefer for phytotherapy before well understanding their chemistry and biological activities.
Assuntos
Doença Hepática Induzida por Substâncias e Drogas/patologia , Extratos Vegetais/análise , Extratos Vegetais/toxicidade , Polygonum/química , Polygonum/toxicidade , Linhagem Celular , Sobrevivência Celular , Cromatografia Líquida de Alta Pressão , Etanol , Hepatócitos/efeitos dos fármacos , Humanos , Limite de Detecção , Reprodutibilidade dos Testes , Solventes , Sais de Tetrazólio , Tiazóis , ÁguaRESUMO
Polygonum multiflorum is one of the most commonly used Chinese medicines. In this study, an effective pressurized water extraction and HPLC method was developed for first simultaneous determination of 8 hydrophilic compounds, including gallic acid, Hypaphorine, Catechin, Proanthocyanidin B1, Epicatechin, Proanthocyanidin B2, Emodin-8-O-ß-d-glucopyranoside, stilbene glycosides, in P. multiflorum. The analysis was performed on a Zorbax SB-AQ column with gradient elution of 0.05% phosphoric acid aqueous solution and acetonitrile in 45 min. All calibration curves showed good linearity (R(2)>0.9994) within test ranges. The LOD and LOQ were lower than 0.2 and 1.0 µg/mL on column, respectively. RSD for intra- and inter-day of 8 analytes were less than 4.1% and 4.0%, respectively, and the overall recovery was 96.0-100.7%. The validated method was successfully applied to quantification of 8 hydrophilic compounds in samples of P. multiflorum from different locations of China. Chemometrics such as principal component analysis (PCA) and hierarchical clustering analysis (HCA) were used to evaluate homogeneity of P. multiflorum in China, which suggested that their quality homogeneity was good.