Up to date, over 700 disinfection byproducts (DBPs) have been detected and identified in drinking water. It has been recognized that cytotoxicity of DBPs varied significantly among groups. Even within the same group, cytotoxicity of different DBP species was also different due to different halogen substitution types and numbers. However, it is still difficult to quantitatively determine the inter-group cytotoxicity relationships of DBPs under the effect of halogen substitution in different cell lines, especially when a large number of DBP groups and multiple cytotoxicity cell lines are involved. In this study, a powerful dimensionless parameter scaling method was adopted to quantitatively determine the relationship of halogen substitution and the cytotoxicity of various DBP groups in three cell lines (i.e., the human breast carcinoma (MVLN), Chinese hamster ovary (CHO), and human hepatoma (Hep G2) cell cytotoxicity) with no need to consider their absolute values and other influences. By introducing the dimensionless parameters D x-orn-species cellline and D¯ x-orn-species cellline , as well as their corresponding linear regression equation coefficients k typeornumber cellline and k¯ typeornumber cellline , the strength and trend of halogen substitution influences on the relative cytotoxic potency could be determined. It was found that the effect of halogen substitution type and number on the cytotoxicity of DBPs followed the same patterns in the three cell lines. The CHO cell cytotoxicity was the most sensitive cell line to evaluate the effect of halogen substitution on the aliphatic DBPs, whereas the MVLN cell cytotoxicity was the most sensitive cell line to evaluate the effect of halogen substitution on the cyclic DBPs. Notably, seven quantitative structure activity relationship (QSAR) models were established, which could not only predict the cytotoxicity data of DBPs, but also help to explain and verify the patterns of halogen substitution effect on cytotoxicity of DBPs., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)