摘要:
目的:采用反相高效液相色谱(RP-HPLC)法对10批不同来源蛇床子药材活性成分进行分离分析,建立评价其质量的特征成分谱。方法:采用十八烷基硅烷键合硅胶柱DiamonsilTMC18色谱柱(150mm×4.6mm,5μm),流动相为乙腈-10 mM醋酸铵水溶液(用醋酸调节pH至4.0,体积比为50:50);紫外检测波长320 nm。结果:以安定为内标,佛手柑内酯、欧前胡素、蛇床子素的回归方程分别为Y=0.068 5X-0.0522,Y=0.0204X+0.023 2,Y=0.057 9X-0.105 8;线性范围分别是4～100μg/mL,4～100μg/mL,10～200μg/mL;平均回收率和RSD分别为98.1%、92.5%、98.4%和4.58%、5.73%、4.51%。结论:本方法简便快速,回收率及重现性良好,适用于蛇床子药材中香豆素类化合物的优化分离及佛手柑内酯、欧前胡素和蛇床子素的定量分析,并为蛇床子药材的质量控制与道地性研究提供了简便的方法。
Abstract:
Objective :To establish a RP-HPLC method for separation and determination of active constituents in Fructus Cnidii from 10 different regions,and evaluate the quality of the Fructus Cnidii. Methods :DiamonsilTM C18(150mm×4.6 mm,5μm) was used with a mobile phase of ammonium acetate buffer(pH 4.0)/acetonitrile(50/50,,v/v) and a flow rate of 1.0 mL/min,at the detection wavelength of 320 nm. Results :The calibration curves were linear in the range of 4～100μg/mL for bergapten(r=0.9998),4～100 μg/mL for imperatorin(r=0.999 6),10～200 μg/mL for osthole(r=0.999 7),respectively.The average recoveries were 98.1%,92.5%,98.4%,with the RSD of 4.58%,5.73%,4.51%,respectively. Conclusion :The method is rapid,simple and accurate,and it can be suitable for the separation and determination of bergapten,imperatorin and osthole in Fructus Cnidii,quality control and geoherbalism research of Fructus Cnidii.
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