目的 建立一种高效准确的提取和分析半边旗中二萜类化合物的方法。方法 以超临界 CO₂ 与改性剂乙醇组成的复合流体萃取半边旗中二萜类化合物,采用正交设计法优化萃取条件;建立了 HPL C-大气压化学电离源(APCI)-MS联用的分析方法,以选择离子监测 (SIM)方式进行含量测定,以选择离子峰面积与总离子流峰面积的比值优选改性剂。主要的二萜类化合物 5 F的准分子离子为 [M-H] -1。采用 [M-H] -1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定量。结果 超临界流体萃取最佳条件 :2 5 MPa、6 0℃、15 %甲醇、流量 3.0 m L /min;分析条件 :Diamonsil ODS色谱柱 (15 0 mm× 4.6 mm,5 μm),流动相为 CH3CN-2 mm ol/ L NH4 AC(35∶ 6 5),流速1.0 m L / m in,进样 5μL,线性范围为 0.0 5～ 2.5μg,检出限达 0.4 ng。 5 F的加样回收率为 97.8% (n=3)。结论 本法提取效率高、检测灵敏度高、选择性好,可应用于半边旗中二萜类抗癌化合物的深入研究和质量标准的制定。

Object To establish a high efficient and reliable method for extraction and analysis of the diterpenoids in Pteris semipinnata L. Methods Supercritical CO 2 fluid modified with alcohol was used to extract the diterpenoids in P. semipinnata, the extracting conditions were optimized by orthogonal design method, and the modifying solvent was investigated through total ions chromatograghy normalization. A quadrupole mass spectrometer coupled with atmospheric pressure chemical ionization interface was employed as a detector for HPLC, the-1 ion was used as selective ion for the detecting of ent 11 α hydroxy 15 oxo kaur 16 en 19 olic acid (5F) in selective ion monitoring (SIM) mode. The peak area of SIM and tolal ion chromatogram (TIC) were used for quantitative determination. As an example of its application, this method was used to determinate the content of 5F as antitumor diterpenoids in P. semipinnata. Results The optimized conditions for supercritical CO 2 fluid extraction are 25 MPa, 60 ℃, 15% methanol, flow rate 3 0 mL/min; analytical column was Diamonsil ODS (150 mm×4 6 mm, 5 μm); the mobile phase of HPLC was CH 3CN 2 mmol/L NH 4AC (35∶65), flow rate 1 0 mL/min, injection volume 5 μL; the standard curve showed good linearity over the range of 0 05-2 5 μg; the limit of detection is 0 4 ng; the recovery is 97 8% (n=3). Conclusion This method is highly efficient, sensitive, and selective, which can be applied to study the antitumor drug of diterpenoids in P. semipinnata and to establish the drug standard.

国家自然科学基金资助项目(3987099)