高效液相色谱法同时测定紫花地丁中6 种活性成分

目的：建立超声辅助萃取-高效液相色谱法分离紫花地丁中秦皮乙素、咖啡酸、芦丁、柚皮素、木犀草素和芹菜素6 种活性成分及其含量测定的方法。方法：采用Phenomenex C18色谱柱（150 mm×4.6 mm，5 μm）分离6 种成分；流动相为甲醇-0.1%醋酸溶液，梯度洗脱；流速0.8 mL/min，紫外检测波长340 nm，柱温35 ℃。结果：6 种成分在15 min内均达到基线分离，线性关系良好（r≥0.999 1，n=6），平均回收率均在102.13%～104.68%（相对标准偏差小于1.20%，n=3）。结论：本实验采用超声波法提取，并对提取条件进行探讨，得出最佳条件为超声时间30 min、超声功率120 W、固液比1∶100（g/mL）、甲醇体积分数60%。同时该研究表明该方法简单、快速、经济、可靠，可用于紫花地丁的品质监控。

Simultaneous Determination of Six Active Components in Viola yedoensis Makino by HPLC

Objective: To establish an analytical method for the simultaneous separation and determination of cichorigenin,
caffeic acid, rutin, naringenin, luteolin and kaempferol in Viola yedoensis Makino by ultrasonic-assisted extraction and
high performance liquid chromatography (HPLC). Methods: The separation of six active components was performed on
a Phenomenex C18 column (150 mm × 4.6 mm, 5 μm) with stepwise gradient elution. The mobile phase was a mixture of
methanol and 0.1% acetic acid at a flow rate of 0.8 mL/min; the UV detection wavelength was 340 nm and the column
temperature was set at 35 ℃. Results: Baseline separation of cichorigenin, caffeic acid, rutin, naringenin, luteolin and
kaempferol was achieved within 15 min. The calibration curves of the six components showed good linear relationship (r ≥
0.999 1, n = 6). The average recoveries of spiked samples were in the range of 102.13%–104.68% (RSD < 1.20%, n = 3).
Conclusions: The optimal conditions for ultrasonic-assisted extraction were determined as follows: ultrasonic time, 30 min;
ultrasonic power, 120 W; solid/liquid ratio, 1:100 (g/mL); and extraction solvent, 60% methanol. This study also showed that
the HPLC method was simple, rapid, economic and reliable and could be used for quality control of Viola yedoensis.