Abstract:

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.

Key words: high performance liquid chromatography (HPLC), Viola yedoensis Makino, ultrasonic-assisted extraction; active components, quantitation