732型阳离子树脂分离纯化吴茱萸总碱及抗乳腺癌研究

Separation and purification of total alkaloids from Evodia rutaecarpa using 732cation exchange resin and investigations on its anti-breast cancer bioactivity

目的研究吴茱萸总碱最佳纯化工艺路线及其抗乳腺癌活性。方法分别采用传统提取纯化和732型阳离子交换树脂法提取纯化吴茱萸总碱,用碘化汞钾试剂对其进行定性分析,并对洗脱剂进行选择。应用MTT实验对所提取的吴茱萸总碱进行抗乳腺癌活性测试。结果与传统提取工艺相比,732型阳离子交换树脂法提取吴茱萸总碱的产率较高(16.18%);732型阳离子树脂纯化法提取吴茱萸总碱的最佳洗脱剂为饱和氯化钠溶液;细胞毒性实验结果表明U251乳腺癌细胞对吴茱萸总碱的浓度依赖性不强,即随着载药浓度的增加,细胞的形态改变不大、凋亡的细胞数量增多不明显。当浓度为50μmol/m L,作用时间为24 h时,细胞的存活率为68%;随着浓度的增加(100、150μmol/m L),作用时间为24 h时,细胞的存活率分别为66%和60%。结论与传统提取法相比,732型阳离子树脂纯化法提取的吴茱萸总碱纯度较高;同时吴茱萸总碱对乳腺癌细胞具有明显的抑制作用。

Objective To optimize conditions for extraction of alkaloids from Evodia rutaecarpa by means of 732 cation exchange resin and performe its anti-breast cancer bioactivity. Methods The optimum processing route on extraction of alkaloids from Evodia rutaecarpa was investigated by means of 732 cation exchange resin. The performance of 732 cation exchange resin was compared with tranditonal process（ aqueous extraction-ethanol precipitation in extraction-purification process）. The alkaloid reagent-potassium mercuric iodide test and MTT test were performed on the crudes. Results Compared with traditional purification process,it was much better to use 732 cation exchange resin approach for extraction of alkaloids from Evodia rutaecarp with high yield（ 16. 81%） The best eluent should be saturated brine with excellent purification. No obvious correlations were found between the toxin of breast cancer cell and concentration of total alkaloids. When the concentration of total alkaloids was 50 μmol / m L,cellular survival rate was 68%afterward 24 h. When the concentration of total alkaloids comes to 100 μmol / m L and 150 μmol / m L,cellular survival rates were slightly decreased by66% and 60%. Conclusion 732 cation exchange resin performes much higher than traditional process in purification of total alkaloids extracted from Evodia rutaecarpa. Simultaneously,the extracted total alkaloids showe remarkable inhibition in breast cancer cell.