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Toxicity bioassays with concentrated cell culture media—a methodology to overcome the chemical loss by conventional preparation of water samples

机译:浓缩细胞培养基的毒性生物测定-一种通过常规水样制备克服化学损失的方法

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摘要

The use of in vitro bioassays for studies of toxic activity in environmental water samples is a rapidly expanding field of research. Cell-based bioassays can assess the total toxicity exerted by a water sample, regardless whether the toxicity is caused by a known or unknown agent or by a complex mixture of different agents. When using bioassays for environmental water samples, it is often necessary to concentrate the water samples before applying the sample. Commonly, water samples are concentrated 10–50 times. However, there is always a risk of losing compounds in the sample in such sample preparation. We have developed an alternative experimental design by preparing a concentrated cell culture medium which was then diluted in the environmental water sample to compose the final cell culture media for the in vitro assays. Water samples from five Swedish waste water treatment plants were analyzed for oxidative stress response, estrogen receptor (ER), and aryl hydrocarbon receptor (AhR) activity using this experimental design. We were able to detect responses equivalent to 8.8–11.3 ng/L TCCD for AhR activity and 0.4–0.9 ng/L 17β-estradiol for ER activity. We were unable to detect oxidative stress response in any of the studied water samples. In conclusion, we have developed an experimental design allowing us to examine environmental water samples in toxicity in vitro assays at a concentration factor close to 1, without the risk of losing known or unknown compounds during an extraction procedure.Electronic supplementary materialThe online version of this article (10.1007/s11356-018-1656-4) contains supplementary material, which is available to authorized users.
机译:使用体外生物测定法研究环境水样品中的毒性活性是一个快速发展的研究领域。基于细胞的生物测定可以评估水样所产生的总毒性,而无论毒性是由已知还是未知的试剂或不同试剂的复杂混合物引起的。当对环境水样品使用生物测定时,通常有必要在施加样品之前先浓缩水样品。通常,水样品被浓缩10至50倍。然而,在这种样品制备中,总是存在样品中化合物损失的风险。通过制备浓缩的细胞培养基,然后将其稀释在环境水样品中,以组成用于体外测定的最终细胞培养基,我们已经开发出另一种实验设计。使用该实验设计分析了瑞典五家污水处理厂的水样的氧化应激反应,雌激素受体(ER)和芳烃受体(AhR)活性。对于AhR活性,我们能够检测到相当于8.8–11.3 ng / L TCCD的响应,对于ER活性,我们能够检测到0.4–0.9 ng / L的17β-雌二醇。我们无法在任何研究的水样品中检测到氧化应激反应。总而言之,我们开发了一种实验设计,使我们能够在毒性测定中以接近1的浓度因子在体外毒性试验中检查环境水样品,而不会在提取过程中损失已知或未知化合物的风险。文章(10.1007 / s11356-018-1656-4)包含补充材料,授权用户可以使用。

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