Abstract

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 (ED10, ED50) 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

Highlights

  • Food packaging can contribute potentially toxic chemicals to food

  • Based on previous publications [17, 33] and our own preliminary work, the most sensitive methods, i.e., appropriate yeast strain, for performing L-yeast estrogen screen (YES) and P-YES were used to analyze chemicals associated with Food contact materials (FCM)

  • Because this study focuses on effects of the bioassay format between L-YES and P-YES, most experiments were done without chromatography

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Summary

Introduction

Food packaging can contribute potentially toxic chemicals to food. In the European Union, chemicals known to be reproductive toxicants are not allowed to be used in plastic food packaging [1], some endocrine-disrupting chemicals are still associated with plastic packaging [2]. Bioassays are potential tools to monitor for bioactive NIAS that can migrate from FCM [3, 5]. A combination of chemical fractionation and (bio)analysis, i.e., effect-directed analysis (EDA), is useful to elucidate the responsible toxicants [6, 7]. One important class of chemicals potentially coming from FCM is (xeno)estrogens, for which there are several types of bioassays. Used are in vitro reporter gene assays that measure the activation of an estrogen receptor [8]. One such assay is the yeast estrogen screen (YES)

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