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.