HPLC法测定不同产地杭白菊中三种抑制血管紧张素转化酶活性的黄酮化合物含量___

目的：建立高效液相色谱法测定不同产地杭白菊中抑制血管紧张素转化酶（ACE）活性的黄酮类化合物。方法：以本实验室从杭白菊中分离获得的三种黄酮化合物单体芹菜苷 (Apigenin 7-O-β-D-glucoside)、木犀草素 7-O-(6″-O-丙二酸单酰)-β-D-葡萄糖苷 (Luteolin 7-O-(6″-O-malonyl)-β-D-glucoside)、芹菜素 7-O-(6″-O-丙二酸单酰)-β-D-葡萄糖苷 (Apigenin 7-O-(6″-O-malonyl)-β-D-glucoside)作为对照品，它们对ACE抑制活性筛选结果表明均有显著抑制作用。用反相高效液相色谱测定不同产地杭白菊中这三种黄酮化合物的含量，采用Agilent zorbax SB-C18色谱柱（4.6 ×150 mm,5 µm）,流动相为0.1 %乙酸溶液-乙腈,梯度洗脱60 min，流速1.0mL• min-1，检测波长330 nm，进样量20 µL。结果：芹菜苷 、木犀草素 7-O-(6″-O-丙二酸单酰)-β-D-葡萄糖苷、芹菜素 7-O-(6″-O-丙二酸单酰)-β-D-葡萄糖苷的线性范围分别为10.9~350 µg• mL-1（r=0.9992），16.8~540 µg• mL-1（r=0.9991），16.2~520 µg• mL-1（r=0.9994）。平均回收率分别为100.2%, 97.8%, 98.2%。日内、日间精密度RSD分别为1.5 %，0.6 % ，3.3 %和1.3 %，2.5 %，2.9 %。结论：此方法操作简便、灵敏、准确，重复性好，适用于杭白菊中具有血管紧张素转化酶抑制作用的黄酮化合物含量测定。

To establish an HPLC method for the determination of flavonoids from different places Chrysanthemum which inhibited angiotensin converting enzyme.Method : Apigenin 7-O-β-D-glucoside, Luteolin 7-O-(6″-O-malonyl)-β-D-glucoside, Apigenin 7-O-(6″-O-malonyl)-β-D-glucoside, were used as the standards which were extracted and identified by our lab.and the compounds inhibiting the activities of angiotensin converting enzyme were determined. Agilent zorbax SB-C18 chromatographic column (4.6 ×150 mm,5 µm）was used and the gradient elution solvent system composed of 0.1 % acetic acid solution-acetonitrile was used as the mobile phase. The flow rate was 1.0 mL•min-1, the detection wavelength was set at 330 nm and the volume of injection was 20 µL. Results : the calibration curve was linear within the range of 10.9~350µg• mL-1（r=0.9992），16.8~540µg• mL-1（r=0.9991），16.2~520µg• mL-1（r=0.9994）respectively. The mean recovery for three kinds of concentration of these three standards were 100.2 %, 97.8 % , 98.2 % respectively. Intra-day and inter-day RSD were 1.5 % , 0.6 % , 3.3 % and 1.3 %, 2.5 %, 2.9 % respectively. Conclusion :The method is simple, sensitive, accurate, reproducible, which is suitable for the determination of angiotensin converting enzyme inhibitory compounds of flavonoids in Chrysanthemum .

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