大孔树脂-填充柱超临界CO_2萃取联用精制冬凌草二萜有效部位

A Protocol for Production of Diterpenoids from Rabdosia rubescens by Combination of Macroporous Adsorption Resin and CO_2 Supercritical Fluid Extraction

目的建立一种有效成分含量高、绿色可行的冬凌草二萜有效部位精制工艺。方法以冬凌草甲素、乙素为指标,通过树脂静态、动态吸附实验优选大孔树脂吸附精制工艺,进一步采用响应面法考察氧化铝填充柱的超临界CO_2萃取精制工艺。结果优化的工艺为以HPD500型大孔树脂对冬凌草进行初步分离,上样药液生药浓度为0.5 g·mL^(-1),上样量为4倍树脂床体积(BV),将吸附后的树脂依次用水20%、40%、60%乙醇各3BV洗脱,收集40%～60%部分;再进行氧化铝填充柱CO2超临界萃取,最佳萃取工艺为夹带剂用量15%,萃取压力40 MPa,萃取温度43.26℃。在此条件下,制备获得的冬凌草二萜有效部位中冬凌草甲素、乙素含量由药材中0.51%提升到63.09%。结论大孔树脂-超临界CO_2柱色谱联用方法,能高效精制冬凌草二萜有效部位,有效提升含量,且工艺绿色、简单、可行,具备较好的生产应用前景。

Objective To develop a green and efficient method for preparation of diterpenoids from Rabdosia rubescens by combination of macroporous resin and CO2 supercritical fluid extraction (SPE). Methods The entire preparation procedure consisted of two simple separat steps. The first step was to investigate the optimal separation method of macroporous resin through the static,dynamic adsorption and desorption experiment using oridonin and ponicidin as indexes. Subsequently, the diterpenoids part of R. rubescens was purified from extraction by SPE through neutral- alumina column using response surface methodology. Results Various macroporous resins were tested to purify diterpenoids and HPD- 500 showed the most suitable adsorption and desorption properties for purification of diterpenoids. The optimized loading concentration,volume and desorption solvent were 0.5g·mL^-1,4 BV,40%-60% ethanol,respectively. The suitable SPE temperature,extaction pressure and modified solvent were 43.26 ℃ ,40 Mpa and 15% ethanol. Under the optimized conditions, the purity of oridonin and ponicidin was increased from 0.51% (raw material) to 62.07% (purified extract). Conclusion The combination of macroporous resin and SPE could improve the internal quality of the diterpenoids active part of R. rubescens as well as shorten the production cycle,thus deserved to be popularized.