Campus Chemical Instrument Center, The Ohio State University, Columbus, Ohio 43210, United States.
Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210, United States.
Anal Chem. 2022 Jun 21;94(24):8674-8682. doi: 10.1021/acs.analchem.2c00891. Epub 2022 Jun 7.
Highly quantitative metabolomics studies of complex biological mixtures are facilitated by the resolution enhancement afforded by 2D NMR spectra such as 2D C-H HSQC spectra. Here, we describe a new public web server, COLMARq, for the semi-automated analysis of sets of 2D HSQC spectra of cohorts of samples. The workflow of COLMARq includes automated peak picking using the deep neural network DEEP Picker, quantitative cross-peak volume extraction by numerical fitting using Voigt Fitter, the matching of corresponding cross-peaks across cohorts of spectra, peak volume normalization between different spectra, database query for metabolite identification, and basic univariate and multivariate statistical analyses of the results. COLMARq allows the analysis of cross-peaks that belong to both known and unknown metabolites. After a user has uploaded cohorts of 2D C-H HSQC and optionally 2D H-H TOCSY spectra in their preferred format, all subsequent steps on the web server can be performed fully automatically, allowing manual editing if needed and the sessions can be saved for later use. The accuracy, versatility, and interactive nature of COLMARq enables quantitative metabolomics analysis, including biomarker identification, of a broad range of complex biological mixtures as is illustrated for cohorts of samples from bacterial cultures of in both its biofilm and planktonic states.
高定量代谢组学研究复杂生物混合物得益于二维 NMR 谱（如二维 C-H HSQC 谱）提供的分辨率增强。在这里，我们描述了一个新的公共网络服务器 COLMARq，用于半自动分析样本队列的二维 HSQC 光谱集。COLMARq 的工作流程包括使用深度神经网络 DEEP Picker 进行自动峰提取、使用 Voigt Fitter 进行数值拟合提取定量交叉峰体积、在光谱队列之间匹配对应交叉峰、在不同光谱之间进行峰体积归一化、进行代谢物鉴定的数据库查询以及对结果进行基本的单变量和多变量统计分析。COLMARq 允许分析属于已知和未知代谢物的交叉峰。在用户以其首选格式上传二维 C-H HSQC 和可选的二维 H-H TOCSY 光谱的队列后，网络服务器上的所有后续步骤都可以完全自动执行，如果需要，可以进行手动编辑，并且可以保存会话以供以后使用。COLMARq 的准确性、多功能性和交互性使包括生物标志物鉴定在内的广泛复杂生物混合物的定量代谢组学分析成为可能，如图所示，对处于生物膜和浮游状态的细菌培养物的样本队列进行了分析。
