HPLC法同时测定川楝子中芦丁、异槲皮苷和槲皮素的含量

目的:采用高效液相色谱法同时测定川楝子药材中芦丁、异槲皮苷和槲皮素的含量。方法:采用Agilent Zorbax SB-C₁₈(4.6 mm×150 mm,5 μm)色谱柱,以甲醇-乙腈(1:10)为流动相A,0.4%磷酸溶液为流动相B,梯度洗脱;流速1.0 mL·min^-1;检测波长360 nm;柱温25℃。结果:芦丁、异槲皮苷和槲皮素浓度在5.0~500.0 μg·mL^-1(r=0.9995)、0.5~50.0 μg·mL^-1(r=0.9999)、1.0~100.0 μg·mL^-1(r=0.9996)范围内与峰面积呈良好的线性关系;平均加样回收率(n=9)分别为96.5%、97.4%、96.3%。所测22份川楝子药材中,芦丁、异槲皮苷和槲皮素的含量范围分别为19.82~120.21、1.43~38.12、4.30~20.48 μg·g^-1。结论:不同购买地川楝子药材中3个黄酮类成分含量差异较大;该方法适用于川楝子中3个黄酮的含量测定。

Objective:To establish an HPLC method for the content determination of rutin,isoquercitrin and quercetin in Fructus Toosendan.Methods:An Agilent zorbax SB-C₁₈ column(4.6 mm×150 mm,5 μm)was adopted.A gradient elution was applied by mixing mobile phase A,which consisted of methanol and acetonitrile(1:10),with mobile phase B which consisted of 0.4% phosphoric acid solution.The flow rate was 1.0 mL·min^-1,the detective wavelength was 360 nm and the column temperature was 25℃.Results:The linear ranges for rutin,isoquercitrin and quercetin were 5.0-500.0 μg·mL^-1(r=0.9995),0.5-50.0 μg·mL^-1(r=0.9999),and 1.0-100.0 μg·mL^-1(r=0.9996),respectively;and the average recoveries (n=9)were 96.5%,97.4%,and 96.3%,respectively.The contents of rutin,isoquercitrin,quercetin and total flavonoids in 22 samples of Fructus Toosendan were in the ranges of 19.82-120.21 μg·g^-1,1.43-38.12 μg·g^-1,and 4.30-20.48 μg·g^-1,respectively.Conclusion:The contents of rutin,isoquercitrin and quercetin in Fructus Toosendan from different origins are quite different.The established method is suitable for the quantitative analysis of rutin,isoquercitrin and quercetin in Fructus Toosendan.