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

Abstract

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.

In vitro genotoxicity of dibutyl phthalate on A549 lung cells at air–liquid interface in exposure concentrations relevant at workplaces

“…Therefore, to further validate if DBP may act as a chemical aneugen and/or clastogen, the respective mechanisms need to be discriminated and a broader range of selected concentrations specifically below the applied DBP concentrations to identify the existence of a benchmark dose would be necessary. In accordance to previously published literature, comet assay seemed more sensitive to detect phthalate induced genetic damage (Al‐Saleh et al, 2017). A critical knowledge gap is the extent of phthalate metabolism in human lungs (Kocbach Bolling et al, 2013), suggesting that inhalation toxicity should incorporate metabolite analysis to reveal the respective genotoxic potential.…”

Biodeterioration of Compost-Pretreated Polyvinyl Chloride Films by Microorganisms Isolated From Weathered Plastics

“…PVC contains large amounts of additives, mostly phthalate esters, and chlorine atoms ( Glas et al, 2014 ; Wen et al, 2014 ). Phthalate esters have been proven to exhibit endocrine disrupting and genotoxic effects ( Al-Saleh et al, 2017 ; Sicinska et al, 2021 ). Similarly, chlorine and its compounds, when reacting with organic molecules, can produce compounds with genotoxic effects ( Richardson et al, 2007 ; Rincon-Bedoya et al, 2013 ).…”

“…Plasticized PVCs contain additives, most often phthalate esters, that improve mechanical properties and durability of the plastic and can represent up to 50% W/W ( Ru et al, 2020 ). Phthalates have been recognized to be endocrine disruptors with genotoxic effects, but other components, such as heavy metals and chlorine atoms, are also of environmental concern in connection with PVC deterioration in the environment and the disposal in landfills ( Glas et al, 2014 ; Wen et al, 2014 ; Al-Saleh et al, 2017 ; Siciňska et al, 2021 ).…”

First Data on PAE Levels in Surface Water in Lakes of the Eastern Coast of Baikal

“…The World Health Organization (WHO) classifies PAEs as endocrine disruptors [6]. The toxicity of PAEs is due to their ability to bind to the human hemoglobin molecule and affect red blood cell function [7], leading to DNA damage [8] and pregnancy termination [9]. Some PAE congeners such as di-(2-ethylhexyl) phthalate (DEHP) and butyl benzyl phthalate (BBP) are considered to be potentially carcinogenic to humans [10].…”