HPLC法测定杜仲-淫羊藿药对中8个化学成分的含量

目的：建立HPLC分析方法同时测定杜仲-淫羊藿药对中绿原酸、咖啡酸、松脂醇二葡萄糖苷、朝藿定A、朝藿定B、朝藿定C、淫羊藿苷、宝藿苷Ⅰ 8个成分的含量，明确配伍前后各成分含量的变化。方法：采用HPLC多波长切换-梯度洗脱技术。色谱柱为Eclipse XDB-C₁₈（4.6 mm&#215;250 mm，5 μm）；流动相为乙腈（A）-0.1%磷酸水溶液（B）；体积流量1.0 mL&#183;min^-1；柱温25℃；进样量5 μL；检测波长：绿原酸、咖啡酸320 nm，松脂醇二葡萄糖苷277 nm，朝藿定A、朝藿定B、朝藿定C 270 nm，淫羊藿苷、宝藿苷Ⅰ280 nm。结果：杜仲-淫羊藿药对中绿原酸、咖啡酸、松脂醇二葡萄糖苷、朝藿定A、朝藿定B、朝藿定C、淫羊藿苷、宝藿苷Ⅰ 8个成分均实现良好分离，进样量分别在4.933&#215;10^-2~1.579 μg、2.554&#215;10^-3~8.172&#215;10^-2 μg、3.957&#215;10^-2~1.266 μg、4.493&#215;10^-2~1.438 μg、5.979&#215;10^-2~1.913 μg、5.108&#215;10^-2~1.635 μg、0.188 0~6.016 μg、1.701&#215;10^-2~0.544 3 μg范围内与峰面积均呈良好的线性关系（r>0.999）；平均加样回收率为97.3%~10³.8%（RSD<3%，n=6）。配伍后绿原酸、咖啡酸、淫羊藿苷、宝藿苷ⅰ的含有量均有不同程度的减少，朝藿定b含有量有不同程度的增加，松脂醇二葡萄糖苷、朝藿定a、朝藿定c含有量无明显变化。结论：本方法快速、稳定、准确、简便，可用于杜仲-淫羊藿药对及其制剂的质量评价与质量控制，并为杜仲、淫羊藿的临床应用及后续配伍研究提供科学依据。

Objective:To establish an HPLC method for simultaneous determination of chlorogenic acid, caffeic acid, pinoresinol diglucoside, epimedin A, epimedin B, epimedin C, icariin and baohuoside Ⅰ in drug pair of Eucommiae Cortex and Epimedii Folium, so as to clear the variation trend in the contents of all ingredients after compatibility.Methods:HPLC multi-wavelength switching and gradient elution was used.Separation was performed on an Eclipse XDB-C₁₈ column (4.6 mm&#215;250 mm, 5 μm) and the mobile phase consisted of acetonitrile (A) -0.1% phosphoric acid aqueous solution (B) at a flow rate of 1.0 mL&#183;min^-1.The column temperature was 25℃ and the injection volume was 5 μL.Detection wavelength was set at 320 nm for chlorogenic acid and caffeic acid, at 277 nm for pinoresinol diglucoside, epimedin A, at 270 nm for epimedin B and epimedin C, and at 280 nm for icariin and baohuoside Ⅰ.Results:Good separation and linearity (r>0.999) were obtained for the 8 ingredients in the range of 4.933&#215;10^-2-1.579 μg、2.554&#215;10^-3-8.172&#215;10^-2 μg、3.957&#215;10^-2-1.266 μg、4.493&#215;10^-2-1.438 μg、5.979&#215;10^-2-1.913 μg、5.108&#215;10^-2-1.635 μg、0.188 0-6.016 μg、1.701&#215;10^-2-0.544 3 μg.The average recovery rates were 97.3%-10³.8% (RSD<3%, n=6).After compatibility, the contents of chlorogenic acid, caffeic acid, icariin and baohuoside Ⅰ reduced by different degrees.The contents of epimedin B increased by different degrees while there was no significant change in the contents of pinoresinol diglucoside, epimedin A and epimedin C.Conclusion:The method is rapid, stable, precise and simple, and can be used to control and evaluate the quality of Eucommiae Cortex and Epimedii Folium;and can provide scientific basis for clinical application and follow-up compatibility study of drug pair of Eucommiae Cortex and Epimedii Folium and their preparations.

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