Abstract:Objective: To establish a reversed-phase high performance liquid chromatographic method for simultaneous determination of 4 coumarins(umbelliferone,daphnoretin-7-O-β-D-glucoside,triumbelletin and daphnoretin) in Wikstroemia indica. Method: Chromatographic separation was performed on an Elite Hypersil-ODS column(4.6 mm×200 mm,5 μm) with a gradient elution of acetonitrile-0.1% phosphoric acid aqueous solution as mobile phase,UV detection wavelength was set at 345 nm,flow rate was 1.0 mL·min-1,column temperature was maintained at 30℃ and injection volumn was 20 μL.Difference of four coumarins in W. indica from nine regions was compared. Result: All the standard compounds showed a good linearity in the range of 0.070-0.770(r=0.999 9),0.190-2.09(r=0.999 9),0.765-8.42(r=0.999 8),4.08-44.9 mg·L-1(r=0.999 9)for umbelliferone,daphnoretin-7-O-β-D-glucoside,triumbelletin and daphnoretin,respectively.Under these conditions,the average recoveries of these four coumarins were 98.0%,96.6%,96.5%,101.3%,respectively and RSD were 1.9%,1.2%,1.7%,1.1%, respectively.Contents of triumbelletin and daphnoretin were significantly higher than umbelliferone and daphnoretin-7-O-β-D-glucoside in W. indica from nine regions,difference between umbelliferone and daphnoretin was small,but difference between daphnoretin-7-O-β-D-glucoside and triumbelletin was big,the content of daphnoretin-7-O-β-D-glucoside was highest from Yunnan region2,Sichuan region was the lowest;The content of triumbelletin was highest from Jiangxi origin,but this ingredient was not detected from Yunnan region1,Guangxi origin3 and Guizhou origin. Conclusion: This developed method was accurate,sensitive and reproducible,it could be successfully applied to quality control of W. indica.
