芫花药材中4个黄酮苷元的定性鉴别定量分析___

Abstract Objective:To develop qualitative identification and quantitative analysis method on 4 flavonoid aglycones of Flos genkwa.Method:Luteolin,apigenin,3′-hydroxygenkwanin and genkwanin were identified by thin-layer chromatography in Flos Genkwa.Toluene-ethyl acetate-formic acid (8∶4∶0.2) was used as developing solvent and the fluorescent of the compounds were obsevered under 365 nm.Their contents were determined by HPLC.Kromasil C₁₈(4.6 mm×250 mm,5 μm) column was used.The mobile phase was methanol-water-glacial acetic acid(60∶40∶0.5) at a flow rate of 0.8 mL·min^-1.The column temperature was 40 ℃.The detection wavelength of luteolin and 3′-hydroxygenkwanin were set at 350 nm,as well as apigenin and genkwanin were set at 338 nm.The record time was 40 min.Results:The linear ranges of luteolin,apigenin,3′-hydroxygenkwanin,and genkwanin were 0.032-0.16 μg(r=0.9994),0.072-0.36 μg(r=0.9995),0.098-0.49 μg(r=0.9995),and 0.15-0.75 μg(r=0.9997),respectively The average recoveries(n=5) were 99.86% with RSD=1.8%,101.0% with RSD＝2.6%,100.3% with RSD=1.8%,and 100.7% with RSD＝2.6%,respectively.By this HPLC method,the total content of four compounds should be equal or over 0.6%,and the content of luteolin,apigenin,3′-hydroxygenkwanin,genkwanin should be equal or over 0.01%,0.2%,0.05%,0.2%,separately.Conclusion:These analytical methods are simple,accurate with good reproducibility.Satisfactory results are obtained for the simultaneous determination of the four main flavonoid aglycones.There are suitable for quality control of Flos Genkwa.