Mohler Rachel E, Dombek Kenneth M, Hoggard Jamin C, Young Elton T, Synovec Robert E
Department of Chemistry, University of Washington, Box 351700, Seattle, Washington 98195, USA.
Anal Chem. 2006 Apr 15;78(8):2700-9. doi: 10.1021/ac052106o.
Comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry coupled with rapid chemometric analysis were used to identify chemical differences in metabolite extracts isolated from yeast cells either metabolizing glucose (repressed (R) cells) via fermentation or metabolizing ethanol by respiration (derepressed (DR) cells). Principal component analysis (PCA) followed by parallel factor analysis (PARAFAC) in concert with the LECO ChromaTOF software located and identified the differences in composition between the two types of cell extracts and provided a reliable ratio of the metabolite concentrations. In this report, we demonstrate the analytical method developed to provide relatively rapid analysis of three selective mass channels (m/z 73, 205, 387), although in principle all collected mass channels could be analyzed. Twenty-six metabolites that differentiate repressed cells from derepressed cells were identified. The DR/R ratio of metabolite concentrations ranged from 0.02 for glucose to 67 for trehalose. The average biological variation of the sample extracts was 31%. This analysis demonstrates the utility and benefit of using PCA combined with PARAFAC and ChromaTOF software on extremely complex samples to derive useful information from complex three-dimensional chromatographic data objectively and relatively rapidly.
采用全二维气相色谱-飞行时间质谱联用快速化学计量分析方法，鉴定从酵母细胞中分离出的代谢物提取物的化学差异，这些酵母细胞要么通过发酵代谢葡萄糖（阻遏（R）细胞），要么通过呼吸作用代谢乙醇（去阻遏（DR）细胞）。主成分分析（PCA）之后是平行因子分析（PARAFAC），并结合LECO ChromaTOF软件，定位并识别了两种细胞提取物之间的成分差异，并提供了代谢物浓度的可靠比值。在本报告中，我们展示了所开发的分析方法，该方法可对三个选择性质量通道（m/z 73、205、387）进行相对快速的分析，尽管原则上所有收集到的质量通道都可进行分析。鉴定出了26种区分阻遏细胞和去阻遏细胞的代谢物。代谢物浓度的DR/R比值范围从葡萄糖的0.02到海藻糖的67。样品提取物的平均生物学变异为31%。该分析证明了在极其复杂的样品上使用PCA结合PARAFAC和ChromaTOF软件，以客观且相对快速地从复杂的三维色谱数据中获取有用信息的实用性和益处。
