紫玉簪活性甾体皂苷的制备工艺研究

Optimal Extraction and Purification Method of the Active Steroidal Saponins from Hosta ventricosa

采用HPLC技术,对紫玉簪中具有抑制5α-还原酶、抗白色念珠菌的活性甾体皂苷进行了制备方法研究。经单因素考察、结合正交实验,确定甾体皂苷最佳提取方法为:紫玉簪根加14倍量水,回流提取2次,每次0.5h,得皂苷提取物。进一步经单因素考察,确定最佳纯化方法为:皂苷提取物按1∶20的量上D101大孔树脂柱,径高比为1∶12,先用40%乙醇洗脱10 BV(床体积)除去水溶性杂质后,80%乙醇洗脱12 BV得粗皂苷;粗皂苷再经D101柱层析,50%乙醇洗脱3 BV后,70%乙醇洗脱3 BV即得精制皂苷。所得精制皂苷纯度高、活性好,其中单一甾体皂苷含量即大于50%,具有显著抑制5α-还原酶活性(IC50=17.3μg/m L)以及抗白色念珠菌作用(IC50=29.1μg/m L)。

An optimal extraction and purification method of the steroidal saponins with 5α-reductase and Candida albicans inhibitory activities from Hosta ventricosa was investigated by HPLC analysis. The single factor study and orthogonal design were performed to analyze the extract solvent,extraction time and times. As the result,the optimal extraction condition was as follows：the dried root was extracted by reflux with water in solid/liquid ratio of 1∶ 14（ w/v） twice（ each0. 5 h） to produce saponin extract. The optimized purification method of saponin extract was as follows：the extract was subjected to D101 macroporous resin column chromatography with a ratio of diameter to high 1∶ 12 and a ratio of sample to resin of 1∶ 20（ w/w）;the column was eluented with 40% alcohol for 10 BV（ bed volume） to remove the water-soluble purities and followed by 80% alcohol elution for 12 BV to afford the crude saponins. The crude saponins were further refined by D101 resin column chromatography which was firstly eluented with 3 BV 50% alcohol to remove the purities and then with 3 BV 70% alcohol to afford the purified saponins. The purity of purified saponins obtained using the optimal method was above 50%. The purified saponins possessed the potent 5α-reductase inhibitory activity（ IC50= 13. 2μg/m L） and anti-C. albicans effect（IC50= 25. 3 μg/m L）.