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Gastric cancer remains one of the most common and deadly malignancies worldwide. Trace elements are gaining increasing interest due to their dual roles as both essential nutrients and potential carcinogens. This study aimed to compare trace element concentrations in gastric cancer tissues with healthy controls. A cross-sectional study was conducted analyzing 27 trace elements in gastric cancer tissues from 73 patients and healthy gastric tissues from 99 controls. Tissue samples were collected from archived paraffin blocks and analyzed using appropriate analytical methods. Statistical analyses included Mann-Whitney U test, ROC curve analysis, and Kruskal-Wallis test. Due to demographic differences between groups, Firth's penalized logistic regression adjusted for age and sex was used to determine the independent diagnostic value of elements. P-values were corrected using the False Discovery Rate (FDR). Significant differences in trace element concentrations were observed between gastric cancer patients and controls. Silver, bismuth, cadmium, cobalt, chromium, gallium, potassium, lithium, nickel, lead, thallium, antimony, vanadium and selenium) were elevated in cancerous tissues (p < 0.05). Conversely, aluminium, boron, iron, indium,manganese, zinc, and mercury were found in lower concentrations in tumor tissues (p < 0.05). After adjusting for age and sex, Selenium and Zinc levels remained significantly lower in cancer tissues, while Copper was significantly higher. These associations persisted independently of demographic variables. This study demonstrates significant changes in trace element profiles between gastric cancer and healthy tissues. The diagnostic potential of trace element profiling and stage-specific variations open new avenues for research and clinical applications in gastric cancer management.