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Objectives: To evaluate the genotoxic risk to hairdressers exposed daily to chemical substances such as hair dyes, waving and straightening preparations and manicurists products by the Comet assay test (single-cell gel electrophoresis). Methods: The Comet assay was performed on blood samples from 69 female hairdressers (36.4 ± 10.7 years old) currently employed in 21 different beauty institutes in São Paulo, Brazil, and on 55 female control blood donors (32.6 ± 10.0 years old) from the São Paulo University Clinical Hospital blood bank. All the control subjects had occupations other than hairdresser. Comet assays were performed by evaluating 100 blood lymphocytes per individual and graded by visual score according to comet tail length. Results: The hairdressers showed a higher frequency of DNA damage revealed by Comet Score (159.8 ± 71) when compared to the control group (125.4 ± 64.1), and the difference was statistically significant by the Students t-test (P = 0.005). Multiple regression analysis showed that in addition to the hairdressers profession, tobacco use contributed to the higher frequency of cells with comets (P  Conclusions: The observed DNA damage could be associated with the hairdressers occupational environment, where different chemicals are chronically manipulated and inhaled. Considering that this profession in many countries, including Brazil, is not officially regulated, more attention should focus on these professionals not only by legislative bodies but also by multidisciplinary teams able to develop and implement risk prevention and control strategies for chemical, physical and biological agents to which hairdressers are exposed. [ABSTRACT FROM AUTHOR]