Background: Physical activity modifies some postprandial responses such as glycemic control, although it is unclear whether this translates into lower postprandial inflammation. Objective: To determine whether postprandial inflammatory markers are lower in active compared to sedentary middle-aged men. Design: Thirteen active and twelve sedentary middle-aged men consumed a mixed meal on one occasion. Blood was taken via a cannula before and up to 8 h after the meal and with a single-use needle before and 8 h after the meal. Results: Active men had lower fasted interleukin-6 (IL-6; 0.6 ± 0.2 pg/ml vs. 1.2 ± 0.3 pg/ml; P = 0.004) and C-reactive protein (CRP; 1.3 ± 0.3 mg/l vs. 2.9 ± 0.6 mg/l; P = 0.04) concentrations than sedentary men. Cannula blood IL-6 concentrations increased by 3.45 pg/ml in the 8 h following the meal (P < 0.001); however, this increase was minimal (0.36 pg/ml) in blood taken via a single-use needle from the contralateral arm (P = 0.013). The sedentary group had larger glucose (P = 0.034), insulin (P = 0.013) and triacylglycerol (P = 0.057) responses to the meal. Conclusions: These results provide further evidence that physical activity is associated with lower inflammatory marker concentrations in a fasted state and a lower postprandial metabolic response to a meal. However, this does not translate into lower postprandial inflammatory markers since the only evidence of postprandial inflammation (a large increase in plasma IL-6) was actually due to the cannula used for blood sampling.