First-order removal rate coefficients (k) are used in predictive equations for estimating effluent concentrations from horizontal flow (HF) wetlands. Due to limited resources, influent and effluent concentration data from existing systems are frequently used in the estimation of k values from operating systems, but another choice is to use concentration data along the longitudinal profile of the HF wetland. Based on a dataset with 41 HF wetlands/studies obtained from a literature survey, with chemical oxygen demand (COD) measurements at different sampling points, volumetric (kV) and areal (kA) removal rate coefficients for the Tanks-In-Series (TIS) model have been obtained using the two estimation methods. In general, removal rate coefficients derived from longitudinal profiles of concentrations were higher than those obtained by using data from influent and effluent concentrations, reflecting the fact that constituent removal is mostly accomplished before the wastewater reaches the outlet zone. Deriving coefficients from longitudinal profiles is more comprehensive, providing a better explanation of the internal removal taking place in the treatment wetland. However, the more widely used approach of calculating kV and kA from influent/effluent concentrations may lead to a safer design of horizontal flow wetlands, because of underestimation of the actual removal rate coefficients.