HPLC-DAD同时测定野菊不同部位8种活性成分的含量

Determination of Eight Active Ingredients in Different Parts of Chrysanthemum Indicum L. by HPLC-DAD

目的建立同时测定中药野菊不同部位(花、叶、嫩茎、老茎)中绿原酸、咖啡酸、1,3-二咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸、木犀草素-7-O-β-D-葡糖苷、芹菜素-7-O-β-D-葡糖苷、蒙花苷和木犀草素含量的高效液相色谱-二极管阵列检测(HPLC-DAD)方法。方法采用Thermo Hypersil C18(250 mm×4.6 mm,5μm)色谱柱,乙腈-0.1%磷酸为流动相,梯度洗脱,流速1.0 ml/min,柱温35℃,检测波长327 nm。结果绿原酸、咖啡酸、1,3-二咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸、木犀草素-7-O-β-D-葡糖苷、芹菜素-7-O-β-D-葡糖苷、蒙花苷和木犀草素进样量分别在0.031~0.620μg、0.009~0.180μg、0.016~0.320μg、0.081~1.620μg、0.021~0.420μg、0.011~0.220μg、0.152~3.040μg、0.017~0.340μg范围内与峰面积呈良好的线性关系,相关系数分别为0.9999,0.9998,0.9997,0.9996,0.9995,0.9996,0.9997,0.9998。绿原酸、咖啡酸、1,3-二咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸、木犀草素-7-O-β-D-葡糖苷、芹菜素-7-O-β-D-葡糖苷、蒙花苷和木犀草素的平均加样回收率分别为98.29%,99.09%,99.03%,98.88%,99.22%,99.02%,98.38%,99.02%,RSD(n=6)分别为1.40%,1.05%,0.98%,1.18%,0.98%,0.91%,0.89%,1.26%。结论本法操作简便,准确性和重复性好,可用于中药野菊的质量控制,测定结果显示野菊不同部位有机酸和黄酮类活性成分含量差异较大。

Objective To develop an HPLC-DAD method for simultaneous determination of chlorogenic acid, caffeic acid, 1,3-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, luteolin-7-O-β-D-glucoside, apigenin-7-O-β-D-glucoside, linarin and luteolin in different parts of Chrysanthemum indicum L.. Methods A Thermo Hypersil C18 column（250 mm×4.6 mm, 5 μm） was used in the gradient elution with a mobile phase of acetonitrile-0.1 % phosphoric acid solution, the flow rate was 1.0 ml/min, the column temperature was 35 ℃ and the detection wavelength was 327 nm. Results Chlorogenic acid, caffeic acid, 1,3-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, luteolin-7-O-β-Dglucoside, apigenin-7-O-β-D-glucoside, linarin and luteolin were linear in the ranges of 0.031-0.620 μg（r=0.9999）, 0.009-0.180 μg（r=0.9998）, 0.016-0.320 μg（r=0.9997）, 0.081-1.620 μg（r=0.9996）, 0.021-0.420 μg（r=0.9995）, 0.011-0.220 μg（r=0.9996）, 0.152-3.040 μg（r=0.9997）, 0.017-0.340 μg（r=0.9998）, respectively. The average recoveries were 98.29 %, 99.09 %, 99.03 %, 98.88 %, 99.22 %, 99.02 %, 98.38 % and 99.02 %, with RSDs of 1.40 %, 1.05 %, 0.98 %, 1.18 %, 0.98 %, 0.91 %, 0.89 % and 1.26 %, respectively. Conclusion The method is convenient, accurate and reproducible, and can be used for quality control of Chrysanthemum indicum L., the results of content determination indicates that the contents of organic acids and flavonoids in different parts of Chrysanthemum indicum L. were different.