HPLC法同时测定藏龙蒿中6种有机酸类成分的含量

Simultaneous determination of 6 organic acids in Artemisia waltonii based on HPLC

目的用高效液相色谱法(HPLC)同时测定民族药藏龙蒿中新绿原酸、绿原酸、隐绿原酸、异绿原酸A、1,5-二咖啡酰奎宁酸、异绿原酸C 6种有机酸成分的含量,为其质量控制提供参考。方法用XBridge C_(18)色谱柱(150 mm×4.6 mm,5μm);乙腈(A)-0.5 mL·L^(-1)磷酸水(B)为流动相进行梯度洗脱;流速为1.0 mL·min^(-1);柱温为30℃;检测波长为327 nm;进样量为5μL。结果新绿原酸、绿原酸、隐绿原酸、异绿原酸A、1,5-二咖啡酰奎宁酸、异绿原酸C的质量浓度分别在2.52~40.31μg·mL^(-1)(r=0.9999)、72.40~1158.37μg·mL^(-1)(r=0.9996)、2.51~40.12μg·mL^(-1)(r=0.9999)、89.38~1430.00μg·mL^(-1)(r=0.9999)、30.18~482.86μg·mL^(-1)(r=0.9995)、19.94~319.01μg·mL^(-1)(r=0.9995)范围内与其各自的峰面积呈现良好的线性关系;精密度、重复性、稳定性实验的RSD值均小于2.00%;平均加样回收率(n=6)分别为107.07%、93.95%、102.34%、99.54%、105.48%、101.71%,RSD值分别为0.40%、1.20%、2.11%、1.25%、1.08%、2.75%。结论该方法快速准确、稳定可靠,可为藏龙蒿质量标准的制定提供借鉴和参考。

Objective To develop a method for the simultaneous determination of neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,isochlorogenic acid A,1,5-dicaffeoylquinic acid and isochlorogenic acid C in Artemisia waltonii based on high performance liquid chromatography(HPLC),and provide reference for quality improvement.Methods The analysis was carried out on XBridge C_(18) column(150 mm×4.6 mm,5μm).Acetonitrile(A)and 0.5 mL·L^(-1) phosphoric acid solution(B)were used as the mobile phase with gradient elution at 1 mL·min^(-1).The column temperature was 30℃.The detection wavelength was 327 nm and the injection volume was 5μL.Results Quantitative analysis showed that the amount of neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,isochlorogenic acid A,1,5-dicaffeoylquinic acid and isochlorogenic acid C showed good linear relationships with the peak area in the ranges of 2.52-40.31μg·mL^(-1)(r=0.9999),72.40-1158.37μg·mL^(-1)(r=0.9996),2.51-40.12μg·mL^(-1)(r=0.9999),89.38-1430.00μg·mL^(-1)(r=0.9999),30.18-482.86μg·mL^(-1)(r=0.9995)and 19.94-319.01μg·mL^(-1)(r=0.9995),respectively.The RSD of precision,repeatability and stability test were all less than 2.00%.The average recoveries(n=6)were 107.07%,93.95%,102.34%,99.54%,105.48%and 101.71%,with the RSD of 0.40%,1.20%,2.11%,1.25%,1.08%and 2.75%,respectively.Conclusion The method is rapid,accurate,stable and reliable,which can provide reference for the development of quality standard of Artemisia waltonii.