半边旗二萜类活性成分的提取分离研究

 目的　建立一种高效的提取和富集分离半边旗中二萜类活性成分的方法。方法 用超临界CO₂与改性剂乙醇组成的复合流体萃取半边旗中二萜类活性成分,采用正交实验法优化萃取条件;采用D101大孔吸附树脂吸附、乙醇阶梯洗脱的方法分离富集半边旗中二萜类活性成分;以高效液相色谱(HPLC)-大气压化学电离源(APCI)-质谱(MS)联用的分析方法跟踪检测,以主要的二萜类活性成分5F为监测目标,采用准分子离子[M-H]^-1为选择离子监测离子,以色谱保留时间和选择监测离子流峰面积定量。结果 超临界流体萃取的最佳条件为25MPa,60℃,15％甲醇,流量3.0mL·min^-1;采用35％乙醇洗脱D101大孔吸附树脂可除极性杂质,45％～55％乙醇洗脱流份可富集分离半边旗中二萜类活性成分。结论　采用超临界CO₂萃取与大孔吸附树脂吸附分离相结合,提取分离半边旗中二萜类活性成分,具有效率高、不使用有毒有害溶剂、环境污染程度低的特点,该方法易于工业化。

Study on extraction and separation of diterpenoids from Pteris semipinnata

OBJECTIVE To develop a HPLC method for extracting and separating the diterpenoids from Pteris semipinnata L.METHODS Supercritical carbon dioxide fluid modified with alcohol was used to extract the diterpenoids in Pteris semipinnata L. .The extracting conditions were optimized by orthogonal design method,and the modified solvent was investigated with total ions chromatogram normalization. The macro-porous absorptive resin with gradient elution by alcohol was used to separate and partial enrich active component 5F.A quadrupole mass spectrometer coupled with atmospheric pressure chemical ionization interface was employed as the detector for tracing and de-terming the content of 5 F in the complicated matrix of products extracted and separated. RESULTS The optimized conditions for supercritical carbon dioxide fluid extraction were: 25 Mpa,60℃,15% methanol.macro-porous absorptive resin with gradient elution separated polar components by 35% alcohol and enrich active component 5F by 45%～55% alcohol. CONCLUSION HPLC-MS is a powerful tool for resolving and detecting the content of 5F in the complicated matrix. It is high-efficient to extract and separate diterpenoids from Pteris semipinnata L by means of supercritical carbon dioxide fluid extraction coupled with macro-porous absorptive resin chromatography, using nonpoisonous and harmless solvents with low environmental pollution,and the method is easy to be industrialized.