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

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.