Tissue matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS)may identify lipids differentially expressed between cancer and adjacent normal tissue. To identify lipidomic profiles for gastric cancer, 24 gastric cancerswere profiled for lipid by the histology-directed, tissue MALDI MS technology. Lipid profiles differed between gastric cancer and adjacent normal tissue samples. At P<0.05, median class prediction accuracy in 100 random training-to-test partitions was 83.3% (5/6) for all classifiers tested. A peak at m/z 741.6 (sphingomyelin 34:1 K) was overexpressed, and a peak at m/z 782.6 (phosphatidylcholine 34:1 Na) was underexpressed in gastric cancers compared with normal tissue. Thus, lipid MALDI MS analysis may capture a global alteration in lipid profile of gastric cancer tissue, distinguishing cancerous epithelium from normal epithelium.