Coke from the Athabasca (Alberta, Canada) oil sands operations may someday be integrated into reclamation landscapes. It is hypothesized that the metals associated with the solid coke may leach into the surrounding environment. Therefore, the main objectives of this study were to characterize the toxicity and chemistry of coke leachates collected from two field lysimeters (i.e. shallow lysimeter and deep lysimeter) over a period of 20 months, as well as from other oil sands coke storage sites. In addition, a batch renewal leaching of coke was conducted to examine the rate of metals release. Chronic toxicity of key metals (e.g. Al, Mn, Ni and V) found in lysimeter coke leachate was evaluated separately. Toxicity test results revealed that whole coke leachates (100% v/v) were acutely toxic to Ceriodaphnia dubia; the 7-day LC50 values were always <25% v/v coke leachate. The deep lysimeter leachate was generally more toxic than the shallow lysimeter leachate, likely because of significantly higher concentrations of vanadium (V) found in the deep lysimeter leachate at all sampling times. Vanadium concentrations were higher than all other metals found in the leachate from both lysimeters, and in the batch renewal leaching study. Furthermore, V found in leachates collected from other oil sands field sites showed a concentration–response relationship with C. dubia survival. Mass balance calculations indicated that 94–98% of potentially leachable V fraction was still present in the coke from two field lysimeters. Evidence gathered from these assessments, including toxic unit (TU) calculations for the elements of concern, suggests that V was the likely cause of toxicity of the deep lysimeter leachate, whereas in the shallow lysimeter leachate both Ni and V could be responsible for the observed toxicity.