Section 303(d) of the Clean Water Act (CWA) requires states to assess and list all streams that do not meet water quality criteria for their designated use classes. In Tennessee, the Tennessee Department of Environment and Conservation (TDEC) uses macroinvertebrate surveys to assess the condition of streams designated for “fish and aquatic life” and the progress of targeted waterbodies toward meeting established standards for sediment. As of yet, no substrate metric has been established to monitor water quality or to document progress toward water quality improvement with respect to fish and aquatic life in Tennessee. A substrate metric that could be efficiently measured and would represent the needs of aquatic species would be valuable for monitoring streams with known sediment impairment to detect water quality improvement. The objectives of this study were to (1) investigate the relationships between riffle substrates and benthic macroinvertebrate data, provided by TDEC; (2) assess the potential use of substrate metrics as a monitoring tool for benthic habitat status; and (3) examine variation in riffle substrates over time in the Ridge and Valley Ecoregion of Tennessee. Bed and interstitial sediment were characterized at sites corresponding with TDEC macroinvertebrate sampling stations. Bed sediment characteristics were significantly correlated with benthic macroinvertebrate data; however, interstitial fines yielded no significant correlations with benthic macroinvertebrate data. Substrate metrics did not differ significantly between varying levels of impairment; however, they did differ significantly when all impaired sites were combined into a single impairment group. The lack of significant differences between varying classes of reach impairment suggests that substrate metrics may not be able to distinguish impairment at the level necessary for monitoring impairment. However, substrate metrics may be of potential use in monitoring sites where impairment is less ambiguous. To investigate change in riffle substrate over time, three sites were monitored over the course of a year. Preliminary observations showed little change in riffle substrate during the study period, suggesting that seasonal restrictions on substrate surveys are unneccessary.