Lipids play important physiological roles in the regulation of cellular homeostasis, including as structural and functional components of cellular membranes, for energy storage, and as intra- and intercellular signaling molecules. Emerging data indicate profound dysregulation of cellular lipid metabolism and signaling in colorectal cancer, and there is an increasing recognition of their contributions to malignancy and metastatic progression. However, a detailed survey of the global ‘lipidomic hallmarks’ of colon cancer, and comprehensive structural characterization of the multitude of isomeric lipid species that may be functionally involved in regulating the interconnected networks and causal relationships between gene, transcript, protein and lipid species on cellular phenotype, is currently lacking. Furthermore, the clinical potential of colon cancer associated lipidomes as diagnostic or prognostic biomarkers of the disease, or as targets of therapeutic intervention, remain largely unknown. Here, to address these needs, I will describe results from our recent studies aimed at the development of mass spectrometry based workflows for comprehensive lipidome analysis, including the characterization of isomeric unsaturated lipids differing only in the locations of their C=C double bonds, and quantification of differences in their abundance between patient matched tumor and normal tissue samples and within a series of molecularly-annotated colon cancer cell lines. The power of integrating transcriptomic, proteomic and lipidomic datasets will also be described for interpreting the functional consequences of these differences in global lipidome inventories, and to determine whether variations in lipidome profiles explain part of the clinical heterogeneity of the disease and constitute a distinct, or complementary, taxonomy relative to currently used genomic and transcriptomic classifications.