HPLC同时测定了哥王中4种香豆素的含量

Simultaneous Determination of Four Coumarins in Wikstroemia indica by HPLC

摘要:

目的: 建立HPLC同时测定了哥王中伞形花内酯、西瑞香素-7-O-β-D-葡萄糖苷、triumbelletin、西瑞香素含量的方法。方法: 采用依利特Hypersil-ODS色谱柱(4.6 mm×200 mm,5 μm),流动相乙腈-0.1%磷酸水溶液梯度洗脱,检测波长345 nm,流速1.0 mL·min^-1,柱温30℃,进样量20 μL。比较9个产地了哥王中4种香豆素含量的差异。结果: 伞形花内酯、西瑞香素-7-O-β-D-葡萄糖苷、triumbelletin、西瑞香素线性范围分别为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),平均回收率分别为98.0%(RSD 1.9%),96.6%(RSD 1.2%),96.5%(RSD 1.7%),101.3%(RSD 1.1%)。不同产地了哥王中triumbelletin和西瑞香素含量明显高于伞型花内酯和西瑞香素-7-O-β-D-葡萄糖苷,伞形花内酯和西瑞香素含量差异较小,西瑞香素-7-O-β-D-葡萄糖苷和triumbelletin含量差异较大,其中云南产地2的西瑞香素-7-O-β-D-葡萄糖苷含量最高,四川产地的含量最低;江西产地的triumbelletin含量最高,云南产地1、广西产地3和贵州产地均未测到triumbelletin。结论: 该方法简便、快速、准确,可作为了哥王药材的质量控制方法。

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.