Background: Anastomotic leakage is a serious complication in gastrointestinal surgery, especially inperitonitis condition. Simple interrupted suture is the gold standard for intestinal anastomosis; however, ithas a greater risk of contamination by peritoneal fluid. Fibrin glue has been used as a barrier to protect theintestinal anastomotic suture by increasing hydroxyproline levels. But its benefit in continuous suture is stilldebatable.Objective: To determine the effect of fibrin glue as a sealent in protecting colon anastomosis with continuoussuture compared to simple interrupted suture in intraperitoneal infection conditions.Method: In this experimental study, the rats were divided into two groups: control group (Groups 1 and 2)and experimental group (Groups 3 and 4). Group 1 and 3 were performed by simple interrupted, while theothers by continuous suture. Full-thickness incisions were made on the left colon and fibrin glue was appliedover the sutures in experimental groups. The samples were taken on day 10.Result: According to hydroxyproline levels (mcg/gr tissue), average values of the groups was 201.18 ±27.81 (Group 1), 208.73 ± 17.81 (Group 2), 245.83 ± 20.47 (Group 3), and 241.55 ± 16.85 (Group 4). Tissuehydroxyproline levels in experimental groups were significantly higher compared to control (P < 0.05). Inexperimental group, mean hydroxyproline levels in subject with continuous suture was lower than simpleinterrupted suture; however it was not significant statistically (p=0.796).Conclusion: As a result, the use of fibrin glue is beneficial to increase hydroxyproline level in colonanastomotic tissue especially in simple interrupted than continuous suture technique.