For acceptance of any chemical agent as an endogenous chemical mediator of inflammation, the agent in question must fulfill some biological requirements which are (a) it should be ubiquitously present in tissues in inactive form, (b) it should be activated during process of inflammation whose increase should be identifiable, (c) it should induce or amplify some events of inflammation, (d) there must be some natural inhibitor of such active form in tissues, (e) it should be able to induce inflammatory reaction after exogenous injection, (f) such reaction should be inhibited by exogenous use of their antagonists, and (g) it should be amplified by use of agonists. Copper in its protein free or protein bound form are reported to act as pathogenic factor in inflammatory processes due to oxidative stress. But their role as endogenous chemical mediator of inflammation does not appear to be investigated thoroughly in light of abovementioned biological criterion of mediator. Present study aims at thorough exploration on role of free copper as endogenous chemical mediator of inflammation in light of above facts. It was done by estimation of total copper, protein-bound copper, and free copper along with estimation of free radical generation, increase in vascular permeability, and cellular infiltration during acute inflammatory reaction induced by carrageenan and concanavalin using chicken skin as test model. It was further evaluated by use of exogenous free copper in experimental model and their subsequent inhibition and amplification by chemical chelators of copper. Present study confirms that free copper fulfilled all the biological requirements for accepting it as an endogenous chemical mediator of inflammation. © 2011 Springer Science+Business Media, LLC.