红葡萄酒中花色苷的超高效液相色谱串联三重四级杆质谱检测方法建立

Determination of anthocyanins in red wine by ultra-high performance liquid chromatography tandem triple quaternary mass spectrometry

采用超高效液相色谱-电喷雾离子化串联三重四级杆质谱联用技术(ultra-high performance liquid chromatography tandem triple quaternary mass spectrometry,UPLC-ESI-QQQ),以葡萄和葡萄酒中5种基本花色苷为标准,利用多反应监测(MRM),确立红葡萄酒中花色苷的分析鉴定方法。结果显示:用色谱柱Poroshell 120 EC-C_(18)(150 mm×2.1 mm,2.7μm),流动相A(V(甲酸)∶V(水)=0.2∶100),流动相B(V(甲酸)∶V(甲醇)∶V(乙腈)=0.2∶50∶50)色谱条件,5种基本花色苷在15 min内得到了很好的分离,其浓度和峰面积呈现良好的线性关系(R^2>0.998 3),且回收率为83.38%～118.71%,日内精密度与日间精密度RSD分别为0.62%～1.70%,2.13%～3.90%,并在28 min内从不同的葡萄酒样品中准确鉴定出45种花色苷,其中包括25种单葡萄糖苷和20种吡喃型葡萄糖苷。该检测方法快速、简单,实现了对红葡萄酒花色苷类物质准确的定性、定量分析。

A method that aimed to identify anthocyanins in red wines was established by high-performance liquid chromatography-electrospray ionization tandem triple quaternary mass spectrometry(UPLC-ESI-QQQ) and multi-reaction monitoring(MRM). Five basic anthocyanins in grapes and wines were used as standards. The results found that the five basic anthocyanins were clearly separated within 15 minutes by Poroshell 120 EC-C18 column. The mobile phase A was formic acid: water(0.2:100, volume fraction) and mobile phase B was formic acid: methanol: acetonitrile(0.2:50:50, volume fraction). Under this condition, a positive linear relationship was established between concentration and peak area(R^2>0.9983). The recovery rate was 83.38%-118.71%. The RSDs of intra-day and inter-day precisions were 0.62%-1.70% and 2.13%-3.90%, respectively. Meanwhile, there were 45 anthocyanins accurately identified from different wine samples within 28 min, which included 25 monoglycosides and 20 pyranoid glucosides. This method is rapid, simple, and accurate, and can be used for qualitative and quantitative analysis of anthocyanins in red wines.