巫山淫羊藿饮片HPLC特征图谱研究及朝藿定C的定量分析

Determination of epimedin C and study on HPLC specific chromatogram of decoction pieces of Epimedium wushanense T.S.Ying

目的:建立巫山淫羊藿饮片中朝藿定C的含量测定方法及巫山淫羊藿饮片的HPLC特征图谱。方法:采用HPLC法对20批巫山淫羊藿饮片进行特征图谱研究,建立HPLC特征图谱共有模式,进行相似度比较,并将巫山淫羊藿饮片与淫羊藿饮片进行相似度比对。色谱条件:采用Welchrom C18(250 mm×4.6 mm,5μm)色谱柱,以乙腈-水为流动相进行梯度洗脱,流速1mL·min-1,检测波长270nm,柱温25℃。结果:朝藿定C进样量在0.0996～4.980μg(r=0.9996)范围内线性关系良好,巫山淫羊藿饮片羊油炙品和生品中的平均加样回收率(n=6)分别为100.7%(RSD=0.64%)和99.78%(RSD=1.5%);建立了巫山淫羊藿的HPLC特征图谱,确定了5个共有峰,巫山淫羊藿饮片生品和羊油炙品的平均相似度为0.91;巫山淫羊藿饮片与淫羊藿饮片的平均相似度为0.84,二者特征图谱差异明显。结论:巫山淫羊藿饮片和淫羊藿饮片特征图谱差异明显,可以作为饮片辨识和鉴别的参考方法。且该方法简便、准确,专属性强,为2010年版中国药典一部增订巫山淫羊藿饮片含量测定项进行了研究。

Objective：To establish an HPLC method for determination of epimedin C and the HPLC specific chromatogram of decoction pieces of Epimedium wushanense T.S.Ying.Methods：HPLC was applied to establish the specific chromatogram of 20 batches of decoction pieces of Epimedium wushanense T.S.Ying,and the similarity of decoction pieces of Epimedium wushanense T.S.Ying and Epimedium brevicornum Maxim.was contrasted.Welchrom C18 column（250 mm×4.6 mm,5 μm） was adopted and a mixture liquid of acetonitrile-water solution as mobile phase in a gradient elution,the flow rate was 1.0 mL·min-1,the detection wavelength was at 270 nm ant the column temperature at 25 ℃.Results：The linear range of epimedin C was 0.9996-4.980 μg（r=0.9996）,the average recoveries of cut crude drug processed and cut crude drug processed with oil of Epimedium wushanense T.S.Ying were 99.78%（RSD=1.5%） and 100.7%（RSD=0.64%）,respectively;The method provided specific chromatogram with excellent separation and 5 common peaks were confirmed,the similarity of cut crude drug processed and cut crude drug processed with oil of Epimedium wushanense T.S.Ying was 0.91,the similarity of decoction pieces of Epimedium wushanense T.S.Ying and decoction pieces of Epimedium brevicornum Maxim.was 0.84,significant difference was observed between them.Conclusion：The specific chromatogram of decoction pieces of Epimedium wushanense T.S.Ying and decoction pieces of Epimedium brevicornum Maxim.have significant difference which can be used as reference method in identification.The method is simple,accurate,creditable and provides a reference for the quality control.