摘要
目的通过灰色关联度分析技术及对Caco-2肠癌细胞增殖情况的影响来评价荆芥总黄酮大孔吸附树脂纯化工艺。方法以总黄酮静态吸附量和解吸率为考察指标筛选大孔吸附树脂型号,采用单因素试验,优化其纯化总黄酮的工艺条件,并采用MTT法测定Caco-2细胞抑制率,同时应用灰色关联度分析技术,计算总黄酮质量分数与对肿瘤细胞增殖抑制率的关联度。结果 HPD-400型大孔吸附树脂对荆芥总黄酮有较好的吸附分离性能,荆芥药材以0.1 g/m L的生药质量浓度、0.8 g/m L(药材/湿树脂)的上柱量上柱,用2倍柱体积(BV)水洗除杂,用8 BV、70%乙醇进行洗脱,体积流量均为4 BV/h,收集洗脱液,挥干,同法进行第2次纯化,总黄酮质量分数为92.15%,对Caco-2细胞增殖抑制率为86.17%,总黄酮质量分数与抑制率灰色关联系数为0.950 2。结论此工艺条件稳定可行,体外抗Caco-2肿瘤细胞药效明确,为开发以荆芥总黄酮纯化物为主成分的抗癌新药奠定基础。
Objective Through the grey correlation analysis technology and the effect of Caco-2 colon cancer cell proliferation to evaluate the optimization of purification process in total flavonoids of Schizonepetae Herba by macroporous adsorption resin. Methods By static adsorption and desorption rate of total flavonoids as examining indexes to screen the ratios of macroporous adsorption resin, using single factor experiment to optimize its purification process conditions of total flavonoids; The Caco-2 cell inhibition rate was determined by MTT method; At the same time, using the grey correlation degree analysis technology, the correlation between the purity of total flavonoids and proliferation inhibition rate of the tumor cell was calculated. Results HPD-400 macroporous adsorption resin has the good adsorption separation performance of the total flavonoids from Schizonepetae Herba, Schizonepetae Herba at the concentration of 0.1 g/m L, 0.8 g/m L（medicine/wet resin） on the amount of column, using 2 BV water to remove impurity, eluting by 9 BV 70% ethanol and all of the flow rate are 4 BV/h, collecting the eluent and dry it; With the same method to do the secondary purification, the purity of total flavonoids is 92.15%, the inhibitory rate of Caco-2 cell proliferation is 86.17%, the greycorrelation coefficient of flavonoids purity and inhibition is 0.950 2. Conclusion This optimization process and the conditions are feasible and stable, the inhibitory efficacy of the in vitro Caco-2 cells is clearly. It lays the foundation of the new anti-cancer drugs based on the purification of total flavonoids in Schizonepetae Herba.
出处
《中草药》
CAS
CSCD
北大核心
2015年第11期1615-1619,共5页
Chinese Traditional and Herbal Drugs
基金
"十二五"重大新药创制科技重大专项(2013ZX09507005)
