超高效液相色谱-线性离子阱/静电场轨道阱高分辨质谱直接分析葡萄酒中38种多酚类化合物

Direct analysis of 38 polyphenols in wine by ultra high performance liquid chromatography-linear ion trap/orbitrap high resolution mass spectrometry

建立了超高效液相色谱-线性离子阱/静电场轨道阱高分辨质谱检测葡萄酒中38种多酚化合物的检测方法。样品过聚醚砜（PES）滤膜后直接上样分析,Hypersil Gold C18色谱柱分离,以乙腈（含0.1%甲酸）和0.1%甲酸水作为流动相梯度洗脱。在m/z 50-1 000范围内进行一级质谱全扫描。以准分子离子峰的精确质量数和提取的色谱图峰面积进行筛查分析和定量,以保留时间和数据依赖扫描（data-dependent scan）模式获得的子离子质谱图进行定性确证。38种多酚化合物的质量偏差不大于5×10^-6（5 ppm）,浓度与特征离子峰面积的线性关系良好（浓度线性范围为两个数量级）,相关系数（R^2）大于0.99,方法检出限为0.002-0.50 mg/kg。3个添加水平的回收率范围为90%-102%,相对标准偏差为0.51%-2.56%。应用该方法检测了葡萄酒中38种多酚化合物的含量,该方法准确、可靠。

Wine has been described previously as a rich source of polyphenols. However,an accurate screening of its complete phenolic profile is still lacking. In the present work,the anal-ysis of 38 polyphenols in wine using ultra high performance liquid chromatography-linear ion trap/orbitrap high resolution mass spectrometry （ UPLC-LTQ/Orbitrap MS ） was explored. Wines were directly detected. The sample was loaded onto a Thermo Hypersil Gold C18 column （100 mm × 2. 1 mm,1. 9 μm）using a gradient elution of acetonitrile/water containing 0. 1%（ v/v）formic acid for the separation. UPLC-LTQ/Orbitrap mass spectrometer acquired full scan MS date for quantification,and data dependent MS2 product ion spectra for identification and/or confirmation. The regression coefficients（R^2）for the calibration curves（two orders of mag-nitude up to the lowest calibration level）in the study were ≥0. 99. The limits of detection for the 38 compounds were 0. 002-0. 50 mg/kg. The average recoveries at three spiked levels were in the range of 90%-102% with the relative standard deviations（ RSDs）of 0. 51%-2. 56%. Mass errors were always ≤ 5 ppm. This procedure was then successfully applied to the analysis of the polyphenols in wines.