Swiss Centre for Applied Ecotoxicology, Eawag, Überlandstrasse 133, 8600, Dübendorf, Switzerland.
Zürich University of Applied Sciences, Grüental 14, 8820, Wädenswil, Switzerland.
Anal Bioanal Chem. 2020 Jul;412(19):4527-4536. doi: 10.1007/s00216-020-02701-w. Epub 2020 May 26.
Food contact materials (FCM) may contain complex mixtures of estrogenic chemicals. A yeast estrogen screen performed on high performance thin-layer chromatography plates (planar-YES, P-YES) is promising for analysis of such mixtures, as it could allow for better elucidation of effects compared with established methods in microtiter plates. However, the P-YES has not been directly compared with established methods. We compared the performance of a microtiter plate YES (lyticase-YES, L-YES) to P-YES on silica gel HPTLC plates using 17β-estradiol (E2), 20 chemicals representative of migrants from plastic FCM, and three migrates of coated metal food cans. Effective doses (ED, ED) and estradiol equivalencies were calculated for each chemical. Thirteen chemicals had calculable EDs in the L-YES or P-YES, with average EDs 13-fold (range 0.63-36) more potent in P-YES than in the L-YES. Normalized to E2, the median estrogenicity was within 1.5-fold (0.43-8.8) between the assays. Therefore, P-YES was as or more sensitive than L-YES but potencies relative to E2 were comparable between assays. With chromatography, the P-YES detected estrogenicity in coated metal cans, effects that were unmeasurable in L-YES. With the sample preparation methods used in this study, both YES assays are sufficiently sensitive to detect bisphenol A below the specific migration limit for plastic packaging (0.05 mg/kg food). This study demonstrates that P-YES outperforms L-YES because it is more sensitive, provides comparable estradiol equivalents, and circumvents confounding mixture effects. The P-YES will be useful for routine monitoring of FCM and toxicant identification in problematic materials. Graphical abstract.
食品接触材料 (FCM) 可能含有复杂的雌激素化学物质混合物。在高效薄层色谱板上进行的酵母雌激素筛选（平面-YES，P-YES）是分析此类混合物的一种很有前途的方法，因为与微滴定板中的既定方法相比，它可以更好地阐明作用。然而，P-YES 尚未与既定方法直接进行比较。我们使用 17β-雌二醇 (E2)、20 种代表塑料 FCM 迁移物的化学物质和三种涂覆金属食品罐的迁移物，比较了微滴定板 YES（溶菌酶-YES，L-YES）和 P-YES 在硅胶 HPTLC 板上的性能。为每种化学物质计算有效剂量 (ED) 和雌二醇当量。在 L-YES 或 P-YES 中，有 13 种化学物质可计算出 ED，P-YES 中的平均 ED 比 L-YES 高 13 倍（范围 0.63-36）。归一化为 E2，两种测定方法的中位数雌激素活性在 1.5 倍以内（0.43-8.8）。因此，P-YES 与 L-YES 一样敏感或更敏感，但相对于 E2 的效力在两种测定方法之间具有可比性。通过色谱法，P-YES 检测到涂覆金属罐中的雌激素活性，而 L-YES 则无法检测到这些活性。使用本研究中使用的样品制备方法，两种 YES 测定法都足够灵敏，可以检测到塑料包装的特定迁移限量以下的双酚 A（0.05 mg/kg 食物）。这项研究表明，P-YES 优于 L-YES，因为它更敏感，提供可比的雌二醇当量，并避免了混杂的混合物效应。P-YES 将有助于对 FCM 的常规监测和问题材料中毒性物质的识别。
