BioDetection Systems BV. , Science Park 406. Amsterdam, The Netherlands.
Environ Sci Technol. 2013 Oct 15;47(20):11651-9. doi: 10.1021/es403810w. Epub 2013 Sep 25.
Polycyclic aromatic hydrocarbons (PAHs) represent a class of ubiquitously occurring environmental compounds that are implicated in a wide range of toxicological effects. Routine measurement of PAH contamination generally involves chemical analytical analysis of a selected group of representatives, for example, EPA-16, which may result in underestimation of the PAH-related toxicity of a sample. Many high molecular weight PAHs are known ligands of the aryl hydrocarbon receptor (AhR), a nuclear receptor that mediates toxic effects related to these compounds. Making use of this property we developed a PAH CALUX assay, a mammalian, H4IIe- cell-based reporter assay for the hazard identification of total PAH mixtures. The PAH CALUX reporter cell line allows for specific, rapid (4 h exposure time) and reliable quantification of AhR-induced luciferase induction relative to benzo[a]pyrene (BaP), which is used as a positive reference PAH congener. Full dose response relationships with inductions over 100-fold were reached within only 2 h of exposure to BaP. The PAH CALUX is highly sensitive, that is, using a 4 h exposure time, a limit of detection (LOD) of 5.2 × 10(-11) M BaP was achieved, and highly accurate, that is, a repeatability of 5.9% and a reproducibility of 6.6% were established. Screening of a selection of PAHs that were prioritized by the European Union and/or the U.S. Environmental Protection Agency showed that the PAH CALUX bioassay has a high predictability, particularly for carcinogenic PAHs. Experiments with synthetic mixtures and reference materials containing complex PAH mixtures show the suitability of the assay for these types of applications. Moreover, the presented results suggest that application of the PAH CALUX will result in a lower risk of underestimation of the toxicity of a sample than chemical analytical approaches that focus on a limited set of prioritized compounds.
多环芳烃（PAHs）是一类普遍存在于环境中的化合物，涉及广泛的毒理学效应。PAH 污染的常规测量通常涉及对一组代表性化合物（例如 EPA-16）进行化学分析，这可能导致对样品中与 PAH 相关的毒性的低估。许多高分子量 PAHs 是芳烃受体（AhR）的已知配体，AhR 是一种介导与这些化合物相关的毒性作用的核受体。利用这一特性，我们开发了一种 PAH CALUX 测定法，这是一种基于哺乳动物 H4IIe 细胞的报告物测定法，用于鉴定总 PAH 混合物的危害。PAH CALUX 报告细胞系允许对 AhR 诱导的荧光素酶诱导进行特异性、快速（4 小时暴露时间）和可靠的定量，相对于用作阳性参考 PAH 同系物的苯并[a]芘（BaP）。在仅 2 小时的 BaP 暴露时间内，即可达到超过 100 倍的诱导的完整剂量反应关系。PAH CALUX 具有很高的灵敏度，也就是说，使用 4 小时的暴露时间，实现了 5.2×10（-11）M BaP 的检测限（LOD），并且具有很高的准确性，即重复性为 5.9%，再现性为 6.6%。对欧盟和/或美国环境保护署优先考虑的一系列 PAHs 进行筛选表明，PAH CALUX 生物测定法具有很高的预测能力，特别是对致癌 PAHs。用合成混合物和含有复杂 PAH 混合物的参考材料进行的实验表明，该测定法适用于这些类型的应用。此外，所提出的结果表明，与侧重于有限数量的优先化合物的化学分析方法相比，应用 PAH CALUX 将降低对样品毒性的低估风险。
