Balk Fabian, Hollender Juliane, Schirmer Kristin
Eawag, Swiss Federal Institute of Aquatic Science and Technology, 8600 Dübendorf, Switzerland; EPF Lausanne, School of Architecture, Civil and Environmental Engineering, 1015 Lausanne, Switzerland.
Eawag, Swiss Federal Institute of Aquatic Science and Technology, 8600 Dübendorf, Switzerland; ETH Zürich, Department of Environmental Systems Science, 8092 Zürich, Switzerland.
Environ Int. 2023 Apr;174:107798. doi: 10.1016/j.envint.2023.107798. Epub 2023 Feb 4.
Permanent rainbow trout (Oncorhynchus mykiss) cell lines represent potential in vitro alternatives to experiments with fish. We here developed a method to assess the bioaccumulation potential of anionic organic compounds in fish, using the rainbow trout liver-derived RTL-W1 cell line. Based on the availability of high quality in vivo bioconcentration (BCF) and biomagnification (BMF) data and the substances' charge state at physiological pH, four anionic compounds were selected: pentachlorophenol (PCP), diclofenac (DCF), tecloftalam (TT) and benzotriazol-tert-butyl-hydroxyl-phenyl propanoic acid (BHPP). The fish cell line acute toxicity assay (OECD TG249) was used to derive effective concentrations 50 % and non-toxic exposure concentrations to determine exposure concentrations for bioaccumulation experiments. Bioaccumulation experiments were performed over 48 h with a total of six time points, at which cell, medium and plastic fractions were sampled and measured using high resolution tandem mass spectrometry after online solid phase extraction. Observed cell internal concentrations were over-predicted by K-derived predictions while pH-dependent octanol-water partitioning (D) and membrane lipid-water partitioning (D) gave better predictions of cell internal concentrations. Measured medium and cell internal concentrations at steady state were used to calculate RTL-W1-based BCF, which were compared to D- or D-based model approaches and in vivo data. With the exception of PCP, the cell-derived BCF best compared to D-based model predictions, which were higher than predictions based on D. All methods predicted the in vivo BCF for diclofenac well. For PCP, the cell-derived BCF was lowest although all BCF predictions underestimated the in vivo BCF by ≥ 1 order of magnitude. The RTL-W1 cells, and all other prediction methods, largely overestimated in vivo BMF, which were available for PCP, TT and BHPP. We conclude that the RTL-W1 cell line can supplement BCF predictions for anionic compounds. For BMF estimations, however, in vitro-in vivo extrapolations need adaptation or a multiple cell line approach.
永久性虹鳟鱼（Oncorhynchus mykiss）细胞系代表了鱼类实验潜在的体外替代方法。我们在此开发了一种方法，利用虹鳟鱼肝衍生的RTL-W1细胞系来评估鱼类中阴离子有机化合物的生物累积潜力。基于高质量的体内生物浓缩（BCF）和生物放大（BMF）数据的可得性以及这些物质在生理pH值下的电荷状态，选择了四种阴离子化合物：五氯苯酚（PCP）、双氯芬酸（DCF）、特氯氟胺（TT）和苯并三唑叔丁基羟基苯基丙酸（BHPP）。使用鱼类细胞系急性毒性试验（经合组织TG249）来得出50%有效浓度和无毒暴露浓度，以确定生物累积实验的暴露浓度。生物累积实验进行了48小时，共六个时间点，在这些时间点采集细胞、培养基和塑料部分的样本，并在在线固相萃取后使用高分辨率串联质谱进行测量。通过基于K的预测对观察到的细胞内浓度进行了过度预测，而pH依赖性正辛醇-水分配系数（D）和膜脂-水分配系数（D）对细胞内浓度的预测效果更好。使用稳态下测量的培养基和细胞内浓度来计算基于RTL-W1的BCF，并将其与基于D或D的模型方法以及体内数据进行比较。除了PCP外，细胞衍生的BCF与基于D的模型预测最为吻合，基于D的模型预测高于基于D的预测。所有方法对双氯芬酸的体内BCF预测都很好。对于PCP，细胞衍生的BCF最低，尽管所有BCF预测都将体内BCF低估了≥1个数量级。RTL-W1细胞以及所有其他预测方法对PCP、TT和BHPP的体内BMF都有很大程度的高估。我们得出结论，RTL-W1细胞系可以补充对阴离子化合物的BCF预测。然而，对于BMF估计，体外-体内外推需要调整或采用多细胞系方法。
