Nutrient stoichiometry within a wetland is affected by the surrounding land use, and may play a significant role in the removal of nitrate (NO 3–N). Tile-drained, agricultural watersheds experience high seasonal inputs of NO 3–N, but low phosphorus (PO 4–P) and dissolved organic carbon (DOC) loads relative to surface water dominated systems. This difference may present stoichiometric conditions that limit denitrification within receiving waterways. We investigated how C:N:P ratios affected denitrification rates of sediments from tile-drained mitigation wetlands incubated for: 0, 5, 10, and 20 days. We then tested whether denitrification rates of sediments from surface-water and tile-drained wetlands responded differently to C:N ratios of 2:1 versus 4:1. Ratios of C:N:P (P < 0.05) and incubation length (P < 0.05) had a significant effect on denitrification in tile-drained wetland sediments. Carbon limitation of denitrification became evident at elevated NO 3–N concentrations (20 mg L −1). Denitrification measured from tile water and surface water wetland sediments increased significantly (P < 0.05) at the 2:1 and 4:1 C:N treatments. The results from both experiments suggest wetland sediments provide a limiting pool of labile DOC to maintain prolonged NO 3–N removal. Also, DOC limitation became more evident at elevated NO 3–N concentrations (20 mg L −1). Irrespective of NO 3–N concentrations, P did not limit denitrification rates. In addition to wetting period, residence time, and maintenance of anaerobic conditions, the availability of labile DOC is playing an important limiting role in sediment denitrification within mitigation wetlands.