Advances in the application of comprehensive two-dimensional gas chromatography in metabolomics

Abstract

Due to excellent separation capacity for complex mixtures of chemicals, comprehensive two-dimensional gas chromatography (GC × GC) is being utilized with increasing frequency for metabolomics analyses. This review describes recent advances in GC × GC method development for metabolomics, organismal sampling techniques compatible with GC × GC, metabolomic discoveries made using GC × GC, and recommendations and best practices for collecting and reporting GC × GC metabolomics data.

Keywords: Animal models; Biospecimens; Biotransformation; Comprehensive two-dimensional gas chromatography (GC × GC); Data reporting; In vitro analyses; Metabolomics; Multitrophic interactions; Sampling.

Fig. 1.

Metabolome data inform the influences of internal and external perturbations on biological systems (dark boxes, left), and have industrial, technological, and medical applications (light boxes, right).

Fig. 2.

There are trade-offs between feasibility (i.e., costs, sample access) and translatability to living organisms for metabolomics experiments conducted with in vitro cultures, animal models, biospecimens collected non-invasively (e.g., urine, breath) or invasively (e.g., tissue biopsy), and in vivo or human studies.

Fig. 3.

PCA score plot of GC × GC metabolomics data of 258 bacterial samples and 472 metabolites, with samples colored based on priority vs. non-priority preparation batches (a), sample storage time (b) or analysis day (c). Peaks were normalized across samples to account for dilution effects, log₁₀-transformed, and mean-centered and scaled to unit variance.