AbstractField surveys were conducted on golf courses reporting the inability of imidacloprid to control white grubs (Coleoptera: Scarabaeidae) when applied preventively. Surveys of five sites with significant past imidacloprid use (>10 years) revealed significantly greater white grub populations in rough‐mown turf following imidacloprid treatment than that of adjacent short‐mown fairways. Additionally, spatial analysis by distance indicEs (SADIE) analyses demonstrated a positive correlation between white grub and thatch spatial patterns. To investigate the impact of thatch on imidacloprid efficacy and translocation throughout the turfgrass plant, greenhouse experiments were conducted using turf with differing thatch levels. Imidacloprid concentrations in soil and plant tissues were measured with high‐performance liquid chromatography (HPLC) and compared to values obtained through an enzyme‐linked immunosorbent assay (ELISA) kit to determine if the latter could be a cost‐effective alternative in future studies. ELISA provided reliable estimates of concentrations of imidacloprid compared to HPLC, with only minor discrepancies noted across different types of treatments and assessment timings. Despite finding higher imidacloprid levels in leaf tissues compared to roots and some differences in concentration across thatch treatments, there was no clear pattern showing that thatch thickness significantly affects imidacloprid penetration or accumulation in plant tissues or soil over time. These findings suggest that factors other than thatch thickness may contribute to the observed field failures of imidacloprid in controlling white grubs. Further research is necessary to identify these factors and optimize the use of imidacloprid in turfgrass pest management strategies.