HPLC法同时测定黄花蒿水提浸膏中青蒿素、青蒿乙素、猫眼草黄素和猫眼草酚D的含量

Simultaneous Determination of Artemisinin,Arteannuin B,Chrysosplenetin and Chrysosplenol-D in the Water Extract of Artemisia annua L.by HPLC

目的建立高效液相色谱(HPLC)法同时测定黄花蒿水提浸膏中青蒿素、青蒿乙素、猫眼草黄素、猫眼草酚D的含量。方法采用Agilent ZORBAX SB-C18(250 mm×4.6 mm,5μm)色谱柱;流动相:乙腈(A)-纯水(B),梯度洗脱;体积流量:0.8 mL·min^(-1);柱温:30℃;进样量:10μL;检测波长:210 nm。结果青蒿素、青蒿乙素、猫眼草黄素和猫眼草酚D进样量分别在1.6088~16.088μg(r=0.9999)、0.0141~0.1414μg(r=1)、0.1851~1.8509μg(r=0.9999)、0.1441~1.4414μg(r=0.9999)范围内与峰面积呈良好的线性关系,平均加样回收率(n=6)分别为102.44%、97.82%、95.07%、95.55%,RSD分别为1.12%、1.44%、1.29%、1.53%。结论该方法简便准确、可靠稳定、重复性好,可用于同时测定黄花蒿水提浸膏中青蒿素、青蒿乙素、猫眼草酚D、猫眼草黄素的含量。

Objective To establish a HPLC method for the simultaneous determination of artemisinin,arteannuin B,chrysosplenetin and chrysosplenol-D in the water extract of Artemisia annua L.Methods The analysis was performed on Agilent ZORBAX SB-C18(250 mm×4.6 mm,5μm)column with a mobile phase of acetonitrile(A)-water(B)and the flow rate of 0.8 mL·min^(-1)in a gradient elution manner.The column temperature was 30℃.The injection volume was 10μL,and the detection wavelength was 210 nm.Results Artemisinin,arteannuin B,chrysosplenetin and chrysosplenol-D were correlated well linearly with peak area in their respective ranges 1.6088-16.088μg(r=0.9999),0.0141-0.1414μg(r=1),0.1851-1.8509μg(r=0.9999),0.1441-1.4414μg(r=0.9999),the average recovery rate(n=6)were 102.44%,97.82%,95.07%,95.55%,and the RSD values were 1.12%,1.44%,1.29%,1.53%.Conclusion This method is convenient and accurate.It has good stability and repeatability,and can be used to simultaneously determine the content of artemisinin,arteannuin B,chrysosplenetin and chrysosplenol-D in the water extract of Artemisia annua L.