Purpose: To report the clinicopathologic correlation and biochemical analysis of silicone oil removed after 23 years in an eye.Methods: A 63-years-old man with a history of HIV/AIDS and cytomegalovirus retinitis, status post retinal detachment repair with silicone oil at the age of 39 years, presented with several weeks of worse vision. He was found to have a shallow fovea-off tractional retinal detachment. After the silicone oil was removed during retinal detachment repair, it was analyzed by Fouriertransform infrared spectroscopy and gas chromatography with mass spectrometry.Results: In addition to cyclic and linear silicone oil, cholesterol was found in the removed silicone oil, which was not present in unused silicone oil samples. No other chemical alterations were identified in the extracted silicone oil.Conclusion: Silicone oil left inside an eye over an extended period may extract lipophilic substances from adjacent tissue, with possible pathophysiologic effects. However, no other major potentially toxic substance was identified from the long-standing silicone oil sample, suggesting relative chemical stability of the tamponade agent over time.