Extraction of diterpenoids from Pteris semipinnata by supercritical CO_2 fluid and their analysis with HPLC-MS / 中草药

ABSTRACT

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 ? 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 ?m); the mobile phase of HPLC was CH 3CN 2 mmol/L NH 4AC (35∶65), flow rate 1 0 mL/min, injection volume 5 ?L; the standard curve showed good linearity over the range of 0 05-2 5 ?g; 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.