Environmental Health Program, King Faisal Specialist Hospital & Research Centre, PO Box: 3354, Riyadh, 11211, Saudi Arabia.
Stem Cell and Tissue Re-Engineering Program, King Faisal Specialist Hospital & Research Centre, PO Box: 3354, Riyadh, 11211, Saudi Arabia.
Environ Sci Pollut Res Int. 2017 Oct;24(30):23903-23914. doi: 10.1007/s11356-017-9978-1. Epub 2017 Sep 5.
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.
我们之前报道了在 47 种品牌的香水中，大量使用邻苯二甲酸酯（PAEs）作为溶剂或固定剂。邻苯二甲酸二乙酯的含量最高（0.232-23649ppm），83.3%的香水含量超过 1ppm，这是绿色和平组织调查引用的阈值。所有样品中都含有较高水平的邻苯二甲酸二甲酯，高于其 0.1ppm 的阈值，88%、38%和 7%的香水含有邻苯二甲酸丁基苄基酯、邻苯二甲酸二（2-乙基己基）酯和邻苯二甲酸二丁酯，均高于其阈值。邻苯二甲酸酯作为内分泌干扰物的作用已得到充分证实，但它们对遗传毒性行为的影响却很少受到关注。我们使用人淋巴母细胞 TK6 细胞的体外单细胞凝胶电泳（彗星）和微核（MN）试验，评估了 42 种相同香水的遗传毒性效力，并确定其与 PAEs 的关系。所有的香水都比阴性对照（NEG）诱导了更多的 DNA 损伤，≥90%的样本比用香水制备过程中可能使用的甲醇（ME）和乙醇（ET）处理的细胞造成了更多的损伤，11.6%的香水比阳性对照（过氧化氢）造成了更多的 DNA 损伤。用 71.4%、64.3%、57.1%和 4.8%的香水处理的细胞的染色体断裂表达为 MN 频率，比 NEG、用 ME 或 ET 处理的细胞和另一个阳性对照（X 射线）的 MN 频率都高。彗星试验和 MN 试验中的遗传毒性反应没有相关性。彗星试验表明，在 0.05 和 0.2ppm 浓度下，5 种 PAEs 单独或混合处理 TK6 细胞，与用香水处理的细胞相比，其损伤没有显著差异。与彗星试验不同，MN 试验对 PAEs 的敏感性在低浓度和高浓度下均较弱，MN 频率通常较低。这项研究首次证明了香水中 PAEs 对 DNA 损伤的可能贡献，并表明应规范其作为溶剂或固定剂的使用。然而，其他具有诱变/遗传毒性的成分也可能导致 DNA 损伤。未来的研究应侧重于应用一系列使用不同细胞模型和不同终点的试验，以确定涉及的遗传毒性机制的范围。
