Background: providing comfort for hospitalized patients is one of the important roles of nurses. The usual method of blood sampling causes pain and increases the likelihood of damage to peripheral veins. Blood sampling of peripheral venous catheter may reduce these complications, but the results of various studies in this regard are contradictory .The aim of this study was to compare the blood sugar of blood samples collected by venipuncture and peripheral catheter. Methods: This cross-sectional, analytical study was conducted on 78 patients admitted to the Poursina Rasht Medical Education Center. Patients were divided into 3 groups. Group 1 received normal saline serum, group 2 received dextrose saline, and group 3 received no infusion. From each patient, two blood samples were taken: One through peripheral venous catheter and one by venipuncture. The blood sugar were examined. Data was analyzed by paired t-test, ANOVA, and Bland–Altman plot. Results: Most of the patients were male (64.1%) and the average age was 51 ± 19 years. There were significant differences in the blood sugar between two methods of venipuncture and blood sampling from catheter in the normal saline group (P=0.0001) ، dextrose saline group (P=0.021) and no infusion group (P=0.017). Conclusion: The result of this study shows that Blood sampling from catheter cannot be used to measure blood sugar level. However, further studies are needed on blood sugar.