果酒中11种酚类化合物反相高效液相色谱检测方法的建立

目的采用反相高效液相色谱法(RP-HPLC)建立11种酚类化合物的检测方法,同时测定其在果酒中的含量。方法色谱柱:Agilent Eclipse XDB C18(4.6mm×250mm,5μm);流动相:梯度洗脱,A相:0.1%磷酸;B相:甲醇,洗脱程序:10%B等度(0～10min);10%～20%B线性变换(10～15min);20%B等度(15～25min);20%～60%B线性变换(25～65min);60%B等度(65～75min);检测波长:210、270、320、327、360nm;柱温30℃;流速1.0ml/min。结果在10～500μg/ml浓度范围内,没食子酸、绿原酸、表儿茶素、阿魏酸、白皮杉醇、金丝桃苷、木犀草素7种功能成分均具有良好的线性相关(R2均≥0.9995);在8～400μg/ml浓度范围内,牡荆素、白藜芦醇、槲皮素、山奈酚4种功能成分均具有良好的线性相关(R2=1.0000);回收率为85.1%～112.0%,RSD均<2%;11种功能成分的定量限为0.32～1.59ng,检测限为0.10～0.48ng。结论建立的方法准确、可靠,实现了11种功能成分的分离分析,可用于其在不同种类果酒中的含量测定。

Simultaneous determination of 11 polyphenols in ratafee by reversed-phase high performance liquid chromatography

Objective To develop a novel quantitative method for simultaneous analysis of 11 polyphenols in ratafee by reversed-phase high performance liquid chromatography.Methods The separation was performed on a Agilent Eclipse XDB C18(4.6mm×250mm,5μm)and the column temperature was set at 30℃.The mobile phase was consisted of 0.1% phosphoric acid(A) and methanol(B),with a linear gradient elution at a flow rate of 1.0ml/min.The gradient program was as follow: 10%B isocratic(0-10min),10%-20% B liner(10-15min),20% B isocratic(15-25min),20%-60% B liner(25-65min),60% B isocratic(65-75min).And detection wavelength was 210,270,320,327 and 360nm.Results The correlation coefficients of the 11 calibration curves were all higher than 0.9995.The scopes of the recovery rate was within the range of 85.1% to 112.0%,with RSDs no more than 2%.The limit of quantification(LOQ) and the detection limit(LOD) were 0.32-1.59 ng and 0.10-0.48 ng,respectively.Conclusion The method is accurate and reliable,and can successfully analyze different ratafee samples from different sources.