HPLC同时测定瑞香狼毒中瑞香素、伞形花内酯和东莨菪内酯的含量

Simultaneous Determination of Daphnetin, Umbelliferone and Scopoletin in Stellera Chamejasme L. by HPLC

关键词:  瑞香狼毒  高效液相色谱法  香豆素  瑞香素  伞形花内酯  东莨菪内酯

Key words:  Stellera chamaejasme L.  HPLC  coumarin  daphnetin  umbelliferone  scopoletin

摘要:

目的 建立HPLC同时测定瑞香狼毒中瑞香素、伞形花内酯和东莨菪内酯含量的方法。方法 样品经无水乙醇回流。采用Synergi 4u Polar-RP 80R柱（4.6 mm×250 mm，5 μm），流动相为甲醇-1%冰醋酸溶液，等度洗脱，流速：1.0 mL·min^-1，检测波长：324 nm，柱温：30℃。结果 瑞香素在0.012~0.250 μg（r=0.999 9）、伞形花内酯在0.149~2.981 μg（r=0.999 9）、东莨菪内酯在0.012~0.239 μg（r=0.999 9）均呈良好的线性关系；瑞香素、伞形花内酯和东莨菪内酯的加样回收率分别为99.0%（RSD=0.94%），98.7%（RSD=0.77%）和98.3%（RSD=1.16%）。结论 3种香豆素成分在35 min内达到基线分离，该方法分析时间短、稳定性和回收率良好，对瑞香狼毒药材的质量控制具有一定的参考价值。

Abstract:

OBJECTIVE To establish an HPLC method for the determination of daphnetin, umbelliferone and scopoletin in Stellera chamaejasme L. METHODS Samples were extracted in absolute ethylalcohol with hot reflux. The HPLC was performed on a Synergi 4u polar-RP 80R column(4.6 mm×250 mm, 5 μm) with methanol-1% HAc as isocratic elution. The flow rate was 1.0 mL·min^-1 and the column temperature was kept at 30℃. The detective wavelength was 324 nm. RESULTS The linear range of daphnetin was 0.012-0.250 μg(r=0.999 9); the linear rang of umbelliferone was 0.149-2.981 μg(r=0.999 9), and the linear range of scopoletin was 0.012-0.239 μg(r=0.999 9). The average recovery of daphnetin, umbelliferone and scopoletin were 99.0%(RSD=0.94%), 98.7%(RSD=0.77%) and 98.3%(RSD=1.16%). CONCLUSION The 3 coumarin reached baseline separation within 35 min. This method has advantages of using little analytic time, a good stability and accuracy of operation, which has some references value for establishing standardization quality control method of Stellera chamaejasme L.