Skip to main content

Evaluating the potential genotoxicity of phthalates esters (PAEs) in perfumes using in vitro assays


We previously reported high levels of phthalate esters (PAEs) added as solvents or fixatives in 47 brands of perfumes. Diethyl phthalate was the most abundant compound (0.232–23,649 ppm), and 83.3% of the perfumes had levels >1 ppm, the threshold limit cited by a Greenpeace investigation. All samples had dimethyl phthalate levels higher than its threshold limit of 0.1 ppm, and 88, 38, and 7% of the perfumes had benzyl butyl phthalate, di(2-ethylhexyl) phthalate, and dibutyl phthalate levels, respectively, above their threshold limits. The role of PAEs as endocrine disruptors has been well documented, but their effect on genotoxic behavior has received little attention. We used in vitro single-cell gel electrophoresis (comet) and micronucleus (MN) assays with human lymphoblastoid TK6 cells to evaluate the genotoxic potency of 42 of the same perfumes and to determine its association with PAEs. All perfumes induced more DNA damage than a negative control (NEG), ≥ 90% of the samples caused more damage than cells treated with the vehicles possibly used in perfume’s preparations such as methanol (ME) and ethanol (ET), and 11.6% of the perfumes caused more DNA damage than a positive control (hydrogen peroxide). Chromosome breakage expressed as MN frequency was higher in cells treated with 71.4, 64.3, 57.1, and 4.8% of the perfumes than in NEG, cells treated with ME or ET, and another positive control (x-rays), respectively. The genotoxic responses in the comet and MN assays were not correlated. The comet assay indicated that the damage in TK6 cells treated with five PAEs at concentrations of 0.05 and 0.2 ppm either individually or as a mixture did not differ significantly from the damage in cells treated with the perfumes. Unlike the comet assay, the sensitivity of the MN assay to PAEs was weak at both low and high concentrations, and MN frequencies were generally low. This study demonstrates for the first time the possible contribution of PAEs in perfumes to DNA damage and suggests that their use as solvents or fixatives should be regulated. Other ingredients with mutagenic/genotoxic properties, however, may also have contributed to the DNA damage. Future studies should focus on applying a series of assays that use different cellular models with various endpoints to identify the spectrum of genotoxic mechanisms involved.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3


Download references


The authors acknowledge the roles of (1) Saudi Food and Drug Authorities in sample collection and (2) Sara Bin Judia and Fareed Mahyoub from the Biomedical Physics Department, King Faisal Specialist Hospital and Research Centre for irradiating the cells.

Author information

Authors and Affiliations


Corresponding author

Correspondence to Iman Al-Saleh.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Responsible editor: Philippe Garrigues

Electronic supplementary material


(DOCX 33.2 kb).


(DOCX 32.3 kb).


(DOCX 61 kb).


(DOCX 31.8 kb).


(DOCX 58 kb).


(DOCX 52.1 kb).

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Al-Saleh, I., Al-Rajudi, T., Al-Qudaihi, G. et al. Evaluating the potential genotoxicity of phthalates esters (PAEs) in perfumes using in vitro assays. Environ Sci Pollut Res 24, 23903–23914 (2017).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:


  • Perfumes
  • Phthalate esters
  • Genotoxicity
  • Comet assay
  • Micronucleus test