基于UPLC-Q-TOF-MS技术分析不同品牌啤酒中非挥发性化学成分的差异

Analysis of Differential Non-volatile Compounds in Different Beer Brands by UPLC-Q-TOF-MS

利用超高效液相色谱-四极杆-飞行时间质谱联用技术(ultra-high performance liquid chromatography quadrupole-time-of-flight mass spectrometry,UPLC-Q-TOF-MS)分析4个品牌19种啤酒样品中非挥发性化学成分,探究不同品牌啤酒中非挥发性化学成分的差异,并寻找差异化合物。采用Agilent ZORBAX Eclipse Plus C18(2.1 mm×50 mm,1.8μm)色谱柱,正离子模式下,以水(含0.1%甲酸)和乙腈(含0.1%甲酸)为流动相进行梯度洗脱。利用MassHunter Profinder软件对原始数据进行分子特征提取(molecular feature extraction,MFE),得到2 477个化合物,其中1 056个化合物具有显著性组间差异(P<0.05,倍数变化≥2)。通过韦恩图推测出4个品牌啤酒中存在11个差异化合物,其中品牌1有4个,品牌3有3个,品牌4有4个,品牌2无差异化合物。进一步利用统计软件对具有显著性差异的化合物进行主成分分析(principal component analysis,PCA)和聚类分析,结果显示4个品牌啤酒分类效果显著。该研究为鉴别不同品牌啤酒以及寻找啤酒中特征标志物奠定了基础,同时为啤酒市场的规范和质量监控提供了科学依据。

The non-volatile chemical components of 19 beer samples from 4 brands were analyzed by ultra-high performance liquid chromatography quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) to investigate the differences of non-volatile chemical components in different brands of beer. The Agilent ZORBAX Eclipse Plus C18 column(2.1 mm×50 mm,1.8 μm)and acetonitrile-water(both containing 0.1% formic acid)were used as the mobile phases with mass spectrometry performed in positive ion mode. Molecular feature extraction(MFE)of the raw data using MassHunter Profinder software yielded 2 477 compounds,of which 1 056 had significant group differences(P<0.05,fold change≥2). Eleven differential compounds were inferred from Venn diagrams of four brands of beer,including four in brand 1,three in brand 3,four in brand 4,and no differential compounds in brand 2. The compounds with significant differences were further subjected to principal component analysis(PCA)and cluster analysis using the statistical software Mass Profiler Professional(MPP).The results showed that the samples strongly clustered by brand. This study lays the foundation for identifying different brands of beer based on characteristic markers in beer as well as providing a scientific basis for the regulation and quality control of the beer market.