ABSTRACT
Objective:To establish an HPLC method for the simultaneous determination of chlorogenic acid , aesculetin, rutin, acacetin-7-O-β-D-glucoside, quercetin and luteolin in the extract of Viola yedoensis Makino.Methods:The HPLC analysis was carried out on a Hypersil ODS C18 column (250 mm ×4.6 mm, 5μm) with 0.1％phosphoric acid (A)-methanol (B) as the mobile phase with gradient elution at the flow rate of 1.0 ml· min-1 .The detection wavelength was 345 nm and the column temperature was 35℃. Results:Good linear relationship was found within the range of 4.3175-172.7000 mg· L-1 for chlorogenic acid, 2.7350-109.4000 mg· L1 for aesculetin, 6.9800-279.2000 mg · L-1 for rutin, 3.7200-148.8000 mg · L-1 for acacetin-7-O-β-D-glucoside, 4.1350-165.4000 mg· L-1 for quercetin, and 3.3950-135.8000 mg · L-1 for luteolin.The average recovery was 99.06％, 98.84％, 98.77％, 99.40％, 98.53％and 98.71％, respectively.The content of chlorogenic acid , aesculetin, rutin, acacetin-7-O-β-D-glucoside, quercetin and luteolin in the extract of Viola yedoensis Makino was 1.6290, 1.1910, 4.5850, 2.2810, 3.1790 and 1.9710 mg· g-1, respectively.Conclusion:The method is accurate and reliable with good reproducibility , which can be used for the quality control of the extract of Viola yedoensis Makino.