蒲公英的乙酸乙酯部位HPLC指纹图谱定性分析和咖啡酸的定量测定

The analytical methods with qualitative fingerprint of ethyl acetate extract and quantitative measurement of caffeic acid in Taraxacum mongolicum Hand.Mazz.

目的 运用指纹图谱技术建立蒲公英的色谱定性和定量方法。方法 采用RP -HPLC方法 ,HypersilBDSC18分析柱 (5μm ,2 5 0mm× 4 .6mm) ;流动相A为乙腈 ,流动相B为磷酸盐缓冲液 (磷酸调pH为 3.7) ;梯度洗脱时间为 0、38、4 0min时 ,A(% )分别为 5 %、75 %、75 % ;柱温 35℃ ;流速 0 .7ml·min-l;检测波长 32 3nm。对蒲公英的乙酸乙酯部位进行指纹图谱定性分析 ,并对有效成分咖啡酸进行定量测定。结果 在色谱条件下 ,11份不同蒲公英药材中 ,乙酸乙酯部位的色谱指纹图谱中可检出 11个相对位置稳定的共有峰 ,通过与标准品的保留时间及紫外光谱比较 ,3号峰和 6号峰分别鉴定为咖啡酸和阿魏酸 ,蒲公英药材中咖啡酸的含量为 0 .0 15 %～ 0 .0 4 0 %。结论 蒲公英乙酸乙酯部位HPLC指纹图谱定性和咖啡酸的定量分析方法具有较强的针对性和准确性 。

OBJECTIVE To establish a qualitative and quantitative RP-HPLC method with fingerprinting technology to control the quality of Taraxacum mongolicum Hand.Mazz.METHODS The conditions of RP-HPLC method were as follows: Hypersil BDS C 18 column ( 4.6 mm×250 mm,5 μm) , the mixture of acetonitrile(A) and potassium phosphate solution(B)(pH 3.7) as mobile phase in gradient mode. The concentrations of solvent A were 5%,75% and 75% at 0,38 and 40 min,respectively.The column temperature was set at 35℃ ,the flow rate was 0.7 ml·min -1 and the detecting wavelength was set at 323 nm.The qualitative fingerprint of ethyl acetate extract and quantitative measurement of caffeic acid were performed above the chromatographic conditions. RESULTS Under the qualitative conditions, 11 commom peaks in RP-HPLC fingerprinting of ethyl acetate extract from 11 different samples could be used as index peaks for qualitative identification. By comparison of the retention time and the on-line UV spectra of chemical standards,peak 3 and 6 were identified as caffeic acid and ferulic acid respectively. In RP-HPLC quantitative analysis, the contents of caffeic acid in Taraxacum mongolicum Hand.Mazz.were 0.015%-0.040%.CONCLUSION The analytical method with qualitative fingerprint and quantitative measurement of caffeic acid can accurately be used to control the quality of Taraxacum mongolicum Hand.Mazz.and its preparations.