红花叶总黄酮提取纯化工艺优化及抗急性肝损伤活性研究

Optimization of extraction and purification process of total flavonoids from Carthamus tinctorius L.leaf and its anti-acute liver injury activity

通过单因素试验和Box-Behnken响应面法优化红花叶总黄酮(total flavonoids from Carthamus tinctorius L.leaf,TFCTLL)的提取、纯化工艺。以总黄酮提取率为评价指标,考察料液比、乙醇浓度、提取时间和提取次数四个提取因素的最佳水平。并采用AB-8大孔树脂对获得的TFCTLL进一步富集纯化。结果确定最佳提取工艺为料液比1∶27、乙醇浓度80%、提取时间65 min、提取次数2次。纯化工艺确定选用AB-8大孔树脂、用95%乙醇洗脱、pH=7,富集纯化后得到的TFCTLL纯度为61.42%。采用UPLC-TOF-Q-MS法共鉴定了TFCTLL中木犀草苷等4个化学成分。以CCl_(4)诱导小鼠急性肝损伤模型,实验结果表明TFCTLL给药组小鼠血清中ALT、AST和TBA含量较模型组显著降低,小鼠肝组织病变程度减轻。证明其具有抗急性肝损伤的药效活性。本研究可为红花非药用部位资源的充分利用提供数据支撑。

The extraction and purification process of total flavonoids from Carthamus tinctorius L.leaf(TFCTLL)were optimized by single factor test and Box-Behnken response surface method.Taking the extraction rate of total flavonoids as the evaluation index,the optimal level of four extraction factors,namely,the solid-liquid ratio,the ethanol concentration,the extraction time and the extraction times.The obtained TFCTLL was further enriched and purified with AB-8 macroporous resin.The optimum extraction conditions were as follows:the solid-liquid ratio was 1∶27,ethanol concentrationwas 80%,the extraction time was 65 min,and the extraction times were twice.The purification process is determined to select AB-8 macroporous resin,elute with 95%ethanol,pH=7,eluted with 95%ethanol,pH=7,and The purity of TFCTLL obtained after enrichment and purification is 61.42%.Four chemical constituents including luteolin-7-glucoside in TFCTLL were identified by UPLC-TOF-Q-MS method.CCl_(4)was used to induce acute liver injury model in mice.The experimental results showed that the contents of ALT,AST and TBA in serum of mice in TFCTLL administration group were significantly lower than those in model group,Lesion degree of liver tissue in mice is reduced,It is proved that it has pharmacodynamic activity against acute liver injury.This study can provide data support for the full utilization of non-medicinal parts of Carthamus tinctorius L..