HPLC法同时测定儿茶青黛复合膜中4个成分的含量

HPLC simultaneous determination of four constituents in Catechu and Indigo Naturalis composite membrane

目的:建立同时测定儿茶青黛复合膜中儿茶素、表儿茶素、靛蓝和靛玉红4个成分含量的HPLC方法,以控制该制剂的质量。方法:采用Agilent ZORBAX SB-C18(4.6 mm×250 mm,5μm)色谱柱,流动相组成为水(A)-甲醇(B),梯度洗脱[0 min,A-B(75∶25);10 min,A-B(70∶30);15 min,A-B(63∶37);20 min,A-B(45∶55);25 min,A-B(40∶60);50 min,AB(40∶60)],流速1.0 mL·min-1,检测波长285 nm,柱温30℃。结果:本方法可在45 min内完成儿茶素、表儿茶素、靛蓝和靛玉红4个成分的分析,且各成分色谱峰之间具有良好的分离度。儿茶素、表儿茶素、靛蓝和靛红的进样量分别在0.1386～1.386μg(r=0.9999),0.02694～0.2694μg(r=0.9997),0.1289～1.289μg(r=0.9997),0.02860～0.2860μg(r=0.9997)范围内与色谱峰面积呈良好的线性关系;加样回收率(n=6)均在95.8%～97.1%之间,RSD均小于2.0%。结论:该方法可用于儿茶青黛复合膜的质量控制。

Objective： To establish an HPLC method for the simultaneous determination of catechin,epicatechin,indigotin and indirubin in Catechu and Indigos Naturalis composite membrane,so as to control the quality of this preparation. Methods： The sample was separated on Agilent ZORBAX SB-C18 （4.6 mm×250 mm,5 μm）with the mobile phase consisting of water（A）-methanol（B）by gradient elution program [0 min,A-B（75：25）;10 min,A-B（70：30）;15 min,A-B（63：37）;20 min,A-B（45： 55）;25 min,A-B（40：60）;50 min,A-B（40：60）] at the flow of 1.0 mL·min-1,UV detection wavelength at 285 nm and the column temperature was 30℃. Results： Experimental results showed that a good separation of catechin,epicatechin,indigotin and indirubin was achieved in 45 min by HPLC method.Cate chin,epicatechin,acaciae indigotin and indirubin had good linearity in the ranges of 0.1386-1.386 μg（r=0.9999）,0.02694-0.2694 μg（r=0.9997）,0.1289-1.289 μg（r=0.9997）,0.02860-0.2860 μg（r=0.9996）,respectively.The average recoveries of four constituents were 95.8% -97.1%,with RSDs≤2.0%. Conclusion： The method can be used for quality control of catechu and indigo naturalis composite membrane.