HPLC法同时测定儿茶青黛复合膜中4个成分的含量

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,5 μm)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,A-B(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0.1386~1.386 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=0.9997)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.