In Vivo Real-Time Omics Laboratory, Institute for Advanced Research, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan.
Department of Legal Medicine & Bioethics, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan.
Anal Chem. 2020 Jun 16;92(12):8514-8522. doi: 10.1021/acs.analchem.0c01271. Epub 2020 May 24.
A new analytical platform called PiTMaP was developed for high-throughput direct metabolome analysis by probe electrospray ionization/tandem mass spectrometry (PESI/MS/MS) using an R software-based data pipeline. PESI/MS/MS was used as the data acquisition technique, applying a scheduled-selected reaction monitoring method to expand the targeted metabolites. Seventy-two metabolites mainly related to the central energy metabolism were selected; data acquisition time was optimized using mouse liver and brain samples, indicating that the 2.4 min data acquisition method had a higher repeatability than the 1.2 and 4.8 min methods. A data pipeline was constructed using the R software, and it was proven that it can (i) automatically generate box-and-whisker plots for all metabolites, (ii) perform multivariate analyses such as principal component analysis (PCA) and projection to latent structures-discriminant analysis (PLS-DA), (iii) generate score and loading plots of PCA and PLS-DA, (iv) calculate variable importance of projection (VIP) values, (v) determine a statistical family by VIP value criterion, (vi) perform tests of significance with the false discovery rate (FDR) correction method, and (vii) draw box-and-whisker plots only for significantly changed metabolites. These tasks could be completed within ca. 1 min. Finally, PiTMaP was applied to two cases: (1) an acetaminophen-induced acute liver injury model and control mice and (2) human meningioma samples with different grades (G1-G3), demonstrating the feasibility of PiTMaP. PiTMaP was found to perform data acquisition without tedious sample preparation and a posthoc data analysis within ca. 1 min. Thus, it would be a universal platform to perform rapid metabolic profiling of biological samples.
开发了一种名为 PiTMaP 的新分析平台，用于通过探针电喷雾电离/串联质谱（PESI/MS/MS）进行高通量直接代谢组学分析，该平台使用基于 R 软件的数据管道。PESI/MS/MS 被用作数据采集技术，应用预定选择反应监测方法来扩展靶向代谢物。选择了 72 种主要与中枢能量代谢相关的代谢物；使用小鼠肝和脑样本优化了数据采集时间，表明 2.4 分钟的数据采集方法比 1.2 分钟和 4.8 分钟方法具有更高的重复性。使用 R 软件构建了一个数据管道，事实证明它可以（i）自动为所有代谢物生成箱线图，（ii）执行主成分分析（PCA）和投影到潜在结构判别分析（PLS-DA）等多元分析，（iii）生成 PCA 和 PLS-DA 的得分和加载图，（iv）计算变量重要性投影（VIP）值，（v）通过 VIP 值标准确定统计族，（vi）使用错误发现率（FDR）校正方法进行显著性检验，以及（vii）仅为显着变化的代谢物绘制箱线图。这些任务可以在大约 1 分钟内完成。最后，将 PiTMaP 应用于两种情况：（1）对乙酰氨基酚诱导的急性肝损伤模型和对照小鼠，以及（2）不同等级（G1-G3）的人脑膜瘤样本，证明了 PiTMaP 的可行性。发现 PiTMaP 能够在大约 1 分钟内完成无需繁琐的样品制备和事后数据分析的数据采集。因此，它将成为一种通用平台，可用于快速进行生物样品的代谢组学分析。
