Objectives: To evaluate ability of estimation of tissue extract fluid (TEF) levels of interleukin (IL)-1β, tumor necrosis factor (TNF)-α and IL-6 for differentiation between antemortem (AM) and postmortem (PM) wounds and for rough determination of wound age. Materials & Methods: The study included 20 patients arrived to Surgical Emergency Department (ED) with severe trauma requiring surgical intervention and died after a known survival interval. Time elapsed since trauma inflection till arrival to ED was determined. Four skin biopsies were obtained from each patient: one AM and 3 PM specimens. Skin samples were homogenized and TEF was used for ELISA estimation of IL-1β, TNF-α and IL-6 levels. Results: AM TEF levels of IL-1β and TNF-α were significantly higher compared to PM levels with significantly higher levels in PM1 and PM3 specimens than PM2 specimens. TEF levels of IL-6 were significantly higher in PM2 compared to AM, PM1 and PM3 specimens with significantly higher levels in AM specimens compared to PM3 specimens. TEF levels of IL-1β and TNF-α showed positive significant correlation with wound vitality. TEF levels of IL-6 showed a positive significant correlation with time lapsed since wound inflection. Only elevated TEF level of IL-1β showed significantly high sensitivity for identification of wound inflected since ≤60 minutes AM Conclusion: TEF levels of IL-1β, TNF-α and IL-6 could differentiate between wounds inflected while victim was alive and PM wounds. TEF level of IL-1β works better for such differentiation and could determine wound inflected within 60 minutes with high sensitivity.