Abstract:

Objective: To establish a high performance liquid chromatography (HPLC) method for separation and
quantification of rutin, taxifolin, coumarin, luteolin, kaempferol and apigenin in Agrimonia pilosa Ledeb by ultrasonic-assisted
extraction. Methods: The separation of six flavonoids was performed on Phenomenex C18 column (150 mm × 4.6 mm,
5 μm) using step gradient elution. The mobile phase was composed of methanol and 0.1% acetic acid at a flow rate of 0.8 mL/min.
The UV detection wavelength was 320 nm and the column temperature was set at 35 ℃. Results: Good separation of
rutin, taxifolin, coumarin, luteolin, apigenin and kaempferol was achieved within 15 min. Calibration curves of all the six
components showed an excellent linear relationship (r ≥ 0.999 1, n = 6). The average recoveries were between 97.78%
and 104.4% with a relative standard deviation (RSD) of < 1.20%. Conclusion: This method, based on ultrasonic-assisted
extraction, is simple, fast, economical and reliable.

Key words: high performance liquid chromatography (HPLC), Agrimonia pilosa Ledeb, ultrasonic-assisted extraction, flavonoids, quantitation