消毒剂混合物对蛋白核小球藻的协同作用动态定量表征
Dynamic and Quantitative Characterization of Synergism within Disinfectant Mixtures on Chlorella pyrenoidosa
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摘要: 随着消毒剂在全球范围内的大量使用，越来越多的消毒剂进入环境中，对环境健康构成了威胁。以3种常见的季铵盐类消毒剂为研究对象，包括双十二烷基二甲基溴化铵(didodecyl dimethyl ammonium bromide, DAB)、苯扎溴铵(benzalkonium bromide, BKB)和度米芬(domiphen bromide, DOM)，采用均匀设计射线法设计消毒剂的三元混合体系，应用基于蛋白核小球藻(Chlorella pyrenoidosa, C. pyrenoidosa)的时间依赖微板毒性分析法(time-dependent microplate toxicity analysis, t-MTA)系统考察了消毒剂及其混合物的毒性效应，采用浓度加和(concentration addition, CA)模型定性评估混合物毒性相互作用，并应用拟合归零法动态定量表征毒性相互作用。结果表明，3种消毒剂对C. pyrenoidosa均具有时间依赖毒性效应，在96 h，3种消毒剂的毒性顺序为DOM>BKB>DAB；时间-浓度-效应三维曲面图适合描述效应在空间浓度分布较为均匀的毒性随时间变化规律，在同一暴露时间，消毒剂三元混合物体系对C. pyrenoidosa具有浓度依赖毒性效应，在同一暴露浓度下，消毒剂三元混合体系对C. pyrenoidosa具有时间依赖毒性效应；依据CA模型，三元混合体系的5条射线在24～96 h均表现出时间依赖协同作用；依据拟合归零法，在24～96 h，随时间的延长，5条射线的最大协同作用强度都在逐渐增强，在96 h，5条射线的最大协同作用强度分别为0.3227、0.3507、0.3105、0.2804和0.2626。与CA模型相比，拟合归零法实现了对毒性相互作用的定量表征，更加清晰地呈现出不同浓度水平的毒性相互作用随时间的动态变化规律，为客观评估环境风险提供方法和数据参考。Abstract: With the widespread use of disinfectants around the world, more and more disinfectants enter the environment, posing a threat to environmental health. Three common quaternary ammonium salt disinfectants, didodecyl dimethyl ammonium bromide (DAB), benzalkonium bromide (BKB) and domiphen bromide (DOM), were selected as the research objects. The toxic effects of single disinfectant and their ternary mixture system designed by the uniform design ray method were systematically investigated by the time-dependent microplate toxicity analysis (t-MTA) based on Chlorella pyrenoidosa (C. pyrenoidosa). The toxicity interaction within mixtures were qualitatively evaluated by the concentration addition (CA) model, and quantitatively and dynamically characterized by a fitting-return-to-zero (FRTZ) method. The results showed that three disinfectants all had time-dependent toxicity on C. pyrenoidosa. At 96 h, the toxicity order of the three disinfectants was DOM > BKB > DAB. The time-concentration- response three-dimensional surface diagram was suitable to describe the toxicity changing of pollutants with relatively uniform effect distribution in the whole concentration space in different time. The toxicity of the ternary disinfectant mixture system had concentration-dependency at the same exposure time and time-dependency at the same exposure concentration. According to CA model, five rays in the ternary mixture system showed time-dependent synergism during 24～96 h. According to the FRTZ method, with the extension of time, the maximum synergistic intensity of five rays gradually increases and the maximum synergistic intensity of five rays were 0.3227, 0.3507, 0.3105, 0.2804 and 0.2626 at 96 h, respectively. Compared with CA model, the FRTZ method realizes the quantitative characterization of the toxicity interaction, and more clearly displays the changing rule of toxicity interaction with time at different concentration levels, which provides a method and data reference for the objective assessment of disinfectants’ environmental risks.
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