用HPLC法同时测定六味安消胶囊中土木香内酯、异土木香内酯、大黄素和大黄酚的含量

Simultaneous determination of alantolactone,isoalantolactone,emodin and chrysophanol in Liuwei'anxiao capsules by HPLC method

目的：建立HPLC法同时测定六昧安消胶囊中土木香内酯、异土木香内酯、大黄素和大黄酚含量的方法。方法：采用HPLC-二极管阵列检测器法（HPLC—DAD），色谱柱：Agilent Eclipse PlusC18柱（250mm×4.6mm，5μm），流动相：乙腈-0．04％甲酸（60：40），流速：1．0mL／min，检测波长：220nm，柱温：25℃，进样量：10μL。结果：土木香内酯、异土木香内酯、大黄素和大黄酚的线性范围分别是：2．46～98．4、2．48～99．2、2．56～102．4和2．58～103．2μg／mL，色谱响应线性关系良好。日内和日间RSD均〈2％；最低检测限分别为1．75、1．31、0．50和0．49μg／mL。稳定性结果为峰面积的RSD分别为1．29％、1．40％、1．30％和0．45％，在48h内稳定。加样回收率结果分别为99．0％、100．6％、98．7％和99．7％（n＝6），RSD分别为1．03％、0．62％、1．36％和0．26％。结论：本法快速、简便，重现性好，精密度高，测定结果准确可靠，可作为该制剂的定量测定方法。

Objective： To establish a method for simultaneous determination of alantolactone,isoalantolactone,emodin and ehrysophanol in Liuwei＇anxiao capsules by high performance liquid chromatography-photodiode array detector （HPLC-DAD） method. Methods： An Agilent Eclipse Plus C18 column （250 min×4.6 mm, 5μm） was used with the mobile phase of acetonitrile-0.04% formic acid （60 ： 40） at a flow rate of 1.0 mL/min. The detecting wavelength was 220 nm, the column temperature was 25 ℃ ,and the injection volume was 10μL. Results： The linear ranges of alantolaetone, isoalantolactone, emodin and chrysophanol were 2.46-98.4, 2.48-99.2, 2.56-102.4 and 2.58-103.2 μg/mL, respectively, with a great linear relationship. For the 4 constituents, the results of intra- and inter-day precision （RSD） at low, middle and high concentrations were all less than 2% ; the limits of quantification were 1.75,1.31,0.50 and 0.49 μg/mL, respectively. The stability results in 48 h showed that the RSDs were 1.29% ,1.40% ,1.30% and 0.45%, respectively. The mean recovery rates were 99.0% ,100.6% ,98.7% and 99.7%（n=6）, RSD were 1.03%, 0.62 %,1.36% and 0.26%, respectively. Conclusion： This method is simple, rapid, reproducible, precise, reliable and accurate. It can be used for the quantification of Liuwei＇anxiao capsules.