HPLC法测定窝儿七中5种成分的含量

Determination of 5 main constituents in Diphylleia sinensis Li.by HPLC

目的建立HPLC测定窝儿七中5种成分含量的方法。方法色谱柱为Cosmosil-C18(250 mm×4.6 mm,5μm),柱温25℃;甲醇-4mL·L^(-1)磷酸溶液梯度洗脱;流速1mL·min^(-1);检测波长300nm。结果 4′-去甲基鬼臼毒素(1)、鬼臼毒素(2)、槲皮素(3)、山柰酚(4)和山荷叶素(5)的线性范围分别为4.5~224.0(r=0.999 2),20.0~1 000.0(r=0.999 4),1.6~78.0(r=0.999 3),4.2~210.0(r=0.999 7)和102.2~256.2(r=0.999 4),平均回收率分别为99.8%(RSD=1.4%),99.8%(RSD=1.7%),99.8%(RSD=1.5%),99.9%(RSD=2.3%)和98.1%(RSD=1.7%)。结论该方法灵敏、准确、重复性好,为窝儿七的质量控制提供参考依据。

Objective To establish a quantitative standard of Diphylleia sinensis Li..Methods The chromatographic separation was performed on a Cosmosil-C18 column（250 mm×4.6 mm,5μm）;the column temperature was 25 ℃.A linear elution of4mL·L^-1 phosphoric acid aqueous solution-methanol was adopted at a flow rate of 1mL·min^-1.The UV detection wavelength was 300 nm.Results 4′-Demethylpodophyllotoxin（1）,podophyllotoxin（2）,quercetin（3）,kaempferol（4）,diphyllin（5）showed good linearity in the range of 4.5-224.0（r=0.999 2）,20.0-1 000.0（r=0.999 4）,1.6-78.0（r=0.999 3）,4.2-210.0（r=0.999 7）and 102.2-256.2（r=0.999 4）,and the average recoveries were 99.8%（RSD=1.4%）,99.8%（RSD=1.7%）,99.8%（RSD=1.5%）,99.9%（RSD=2.3%）and 98.1%（RSD=1.70%）,respectively.Conclusion The method is sensitive,accurate and reproducible.It can be used for the determination of main constituents in Diphylleia sinensis Li.