高效液相色谱法测定厚朴中7种代谢产物含量

目的]采用高效液相色谱（HPLC）法同时测定厚朴（Magnolia officinalis Rend.et Wils.）中7种代谢产物含量。方法]色谱条件：Shim-pack CLC-ODS色谱柱（150mm×6mm,5μm）;检测波长：320和294nm;流动相：A相为乙腈,B相为0.1%甲酸,采用梯度洗脱。结果]峰面积（y）对绿原酸、芦丁、金丝桃苷、槲皮苷、槲皮素、厚朴酚和和厚朴酚对照品溶液浓度（x）的回归方程分别为：y=1E＋05x-306360（r=0.9998）、y=5E＋04x-79920（r=0.9994）、y=5E＋04x-55280（r=0.9999）、y=5E＋04x-60530（r=0.9999）、y=2.5E＋04x-57010（r=0.9998）、y=2E＋04x＋7102（r=0.9999）和y=2E＋04x＋11028（r=0.9999）,各对照品溶液在3.125～100.00μg/ml内与峰面积有良好的线性关系。绿原酸、芦丁、金丝桃苷、槲皮苷、槲皮素、厚朴酚和和厚朴酚对照品的加样回收率分别为103.98%、101.65%、99.17%、103.59%、103.59%、102.44%、104.62%,RSD分别为3.37%、2.86%、3.02%、2.89%、2.86%、1.18%、3.67%。厚朴中各部位7种化合物含量存在一定的差异,在叶中,7种代谢产物均有积累,其中和厚朴酚、厚朴酚和槲皮苷的含量较高,金丝桃苷和槲皮素的含量较低。结论]该研究建立的HPLC法简捷、方便,稳定性和重复性好,可为进一步开发利用厚朴提供依据,并为同时测定黄酮类化合物及酚类化合物提供新的方法。

Simultaneous Determination of Seven Metabolites of Magnolia officinalis Rend. et Wils. by High Performance Liquid Chromatography Method

Objective] The research aimed to simultaneously determine seven metabolite contents of Magnolia officinalis Rend. et Wils. by HPLC method. Method] The chromatogram conditions were as follows：Shim-pack CLC-ODS chromatographic column,detective wavelength was 320 and 294nm,mobile phase was phase A acetonitrile and phase B 0.1% formic acid,the velocity was seted according to the setted gradient condition. Result] The concentrations of chlorogenic acid,rutin,hyperin,querctrin,quercetin,magnolol and honokiol liquid （ x ） determined by HPLC method had good linear relationship with the peak areas（ y ） when their concentrations were at 3.125-100.00 μg/ml,and their formulas were y =1E＋05 x-306 360（ r =0.999 8）、 y =5E＋04 x-79 920（ r =0.999 4）、 y =5E＋04 x-55 280（ r =0.999 9）、 y =5E＋04 x-60 530（ r =0.999 9）、 y =2.5E＋04 x-57 010（ r =0.999 8）、 y =2E＋04 x ＋7 102（ r =0.999 9） and y =2E＋04 x ＋11 028（ r =0.999 9） respectively. The average recoveries of chlorogenic acid,rutin,hyperin,querctrin,quercetin,honokiol and magnolol samples were 103.98%,101.65%,99.17%,103.59%,103.59%,102.44% and 104.62%,and their RSD were 3.37%,2.86%,3.02%,2.89%,2.86%,1.18% and 3.67%. The contents of chlorogenic acid,rutin,hyperin,querctrin,quercetin,honokiol and magnolol in Magnolia officinalis Rend. et Wils. had some difference. And the 7 metabolites were all existed in its leaf,thereinto,the contents of honokiol,magnolol and querctrin were high,while hyperin and quercetin were low. Conclusion] The method established by this study was simple,convenient,feasible and repeatabl,can provide basis for the further development and utilization of Magnolia officinalis Rend. et Wils..