Determination of Kaempferol and Quercetin in Botrychium ternatum by HPLC / 医药导报

ABSTRACT

Objective To establish a method for simultaneous determination of kaempferol and quercetin in Botrychium ternatum ( a kind of Ethnic Chinese herbal medicine) by HPLC and HPLC-MS/MS. Methods Agilent 1200 HPLC, API3200 QTRAP MS/MS, Ecilipse XDB-C18 column (15 mm×4.6 mm, 5μm) and the mobile phase consisting of methanol-0.1%formic-water (85??15) were applied to identify kaempferol and quercetin in Botrychium ternatum;Agilent 1260 HPLC and Eclipse plus C18(100 mm×4.6 mm, 5 μm) column were used for determination of the two.The mobile phase was a mixture of methanol and 0.4% phosphoric acid solution (45??55) with a flow rate of 0.8 mL??min-1 and the detection wavelength was 360 nm. Results In mass spectrogram of Botrychium ternatum, characteristic peaks of m/z 287. 1 [ M+H]+ and m/z 301.0 [ M+H]+ were consistent with those of standard sample of kaempferol and quercetin, which confirmed their existence in Botrychium ternatum.The two flavonoids both had a good linearity in the range of 2.500–25.00μg??mL-1(r = 0.999 9 and 0.999 8, respectively).RSD of precision and reproducibility of kaempferol and quercetin were 0.98%,1.89% and 0.79%,2.70%, respectively. Conclusion The method is simple, accurate with high sensitivity and suitable for the determination of kaempferol and quercetin in Botrychium ternatum.