Assays of malathion content and toxicity to boll weevil, Anthonomus grandis grandis Bohemian, were conducted on boll weevil bait sticks, now marketed as Boll Weevil Attract and Control Tubes (BWACTs; Plato Industries, Houston, TX). In general, the longer BWACTs were in the field, the lower the mortality of weevils that were exposed to them. Bioassays of weevil mortality correlated with hexane washes of BWACT surfaces showed highly variable mortality when surface malathion fell below approximately 20 ng per 1 microl of hexane, but consistently high mortality (> or = 90%) when surface malathion was above 30 ng per 1 microl of hexane. A linear equation was calculated to predict mortality as a function of malathion on a BWACT surface. Although mortality was related to surface amounts of malathion, it was unrelated to the total amount of malathion present in BWACTs. Similarly, surface malathion was unrelated to the total amount present in BWACTs. As with mortality, amount of surface malathion declined with time, but total malathion did not decline with time. Boll weevils placed on fresh BWACTs tended to accumulate more malathion and died in greater numbers as time spent on fresh tubes increased, but not as time spent on tubes aged in the field (for 5 mo total) increased. Weevils that landed on tubes after a short flight died in approximately the same numbers as those that were placed on tubes using proper methodology. The amount of malathion expected to cause 90% mortality of boll weevils subjected to proper methodology was 47% higher than for a less stringent methodology (34.3 versus 23.4 ng), which demonstrates the importance of strictly adhering to proper methodology; nevertheless, chemical assay of malathion on the BWACT surface proved to be a more consistent measure of BWACT toxicity than bioassay, and it should replace the bioassay.