In order to reduce the variation of Photobacterium phosphoreum luminous intensity during testing, the Photobacterium phosphoreum toxicity test was modified by a correction factor, PIacetone, in this paper. The effects of magnetic stirring time, culture time, culture generation and culture temperature on the measured results were discussed. The modified method had good laboratory repeatability and the relative standard deviation (RSD) was between 2.1% and 13.1%. Reliable data could be obtained using the modified method. It would be beneficial to the development of the quantitative structure-activity relationships on multicomponent organic compounds.