Seven selected insecticides were applied to three substrates and bioassayed for barrier efficacy and toxicity against the Formosan subterranean termite, Coptotermes formosanus Shiraki, originating from colonies that differed in their tolerance to the insecticides. A profound substrate effect was seen with all insecticides tested. Sand was the most difficult substrate to penetrate and caused faster and greater mortality of C. formosanus. Increased tolerance in C. formosanus was accompanied by a decrease in penetration of treated substrata with chlordane, methoxychlor, chlorpyrifos, and deltamethrin. The opposite was true in the case of permethrin and bendiocarb. More tolerance C. formosanus displayed decreased mortality in the presence of chlorpyrifos, deltamethrin, bendiocarb, and fipronil. The opposite was true for permethrin.