液质联用对半边旗二萜类活性成分分离富集方法的研究

Separation and Enrichment of the Active Diterpenoid Constituent from Pteris Semipinnata L by HPLC-MS

运用高效液相色谱 -质谱联用技术研究了以大孔吸附树脂富集分离 ,并以AgNO3-硅胶柱分离纯化半边旗二萜类活性成分的方法。采用[M -H]-为选择监测离子 ,以色谱保留时间和选择监测离子流峰面积跟踪监测目标成分 ,探讨最佳分离条件 ;以萃取产物的总离子流图 (TIC)和质谱图评价提取产物的纯度。研究表明 ,大孔吸附树脂对5F和4F具有一定的分离富集作用 ;AgNO3-硅胶柱可以分离纯化5F。本实验对复杂基体中化学结构上仅相差1个双键、相对分子质量相差2的5F和4F(无紫外吸收 )进行了分析鉴定 ,获得满意结果。

A method for the separation and enrichment of the active constituent5F(ent-11α-hydroxy-15- oxo-16-kauren-19-oic acid)from pteris semipinnata L using HPLC-MS is described.Macro-porous adˉsorption resin and step-gradient elution with alcohol were employed to separate and partially collect the acˉtive component.Silica gel impregnated with AgNO 3 solution was used for further purification of5F.A quadrupole mass spectrometer coupled with an atmospheric-pressure chemical-ionization interface served as the HPLC detector for the detection and identification of the separated constituents.The retentive time of the SIMion[M-H] - was monitored for constituent identification,whereas the peak area of the SIM ion was measured for its quantitative determination.Both measures were employed to track the target conˉstituent and to find the optimal separation conditions.Total ion chromatograms and mass spectra were meaˉsured to estimate the purity of the extracted product.The successful identification and determination of two similar compounds5F and4F(ent-11α-hydroxy-15-oxo-kaur-19-oic acid),of which the latter comˉpound has no absorption bands in the UV range and differs from the former only by2units in molecular mass and by one double bond in chemical structure,has fully demonstrated the superior resolving power and sensitivity of the HPLC-MS technique.