In order to test various drugs and possibly hazardous compounds on living cells in vitro a system with human spermatozoa was employed. A population of human spermatozoa was transferred into a defined medium by a swim-up procedure or by separation on a Percoll gradient. Such a population is rather homogenous with respect to motility characteristics and was found to be useful for this purpose. Different modes of response were recorded, indicating various effect mechanisms. Effects of various phthalates used as plastic softeners in the production of medical equipment, and extracts from diesel particulate material were recorded. All these compounds interfered with sperm motility in a dose-response fashion. Immediate effects of phthalates were modest, but upon prolonged exposure effects became more evident. Sperm motility was more affected by diethyl-hexyl and dibutyl phthalates. Significant effects were noted for the different phthalates with regard both to percent motility and to some of the various qualities of motility, such as velocity, linearity and amplitude of the track. Thus, the pattern of response considering the motion variables was not the same with the different phthalates. With regard to the effects on sperm motion di-n-octyl phthalate seemed to be the least toxic, followed by dibutyl phthalate. The initial effects of diesel particulate extracts were moderate and mainly restricted to percent motile sperm but upon exposure for 18 hr the effects became more pronounced for all the movement variables.