Validation and application of a robust yeast estrogen bioassay for the screening of estrogenic activity in animal feed

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Abstract

Previously we described the construction and properties of a rapid yeast bioassay stably expressing human estrogen receptor ¿ (hER¿) and yeast enhanced green fluorescent protein (yEGFP), the latter in response to estrogens. In the present study this yeast estrogen assay was validated as a qualitative screening method for the determination of estrogenic activity in animal feed. This validation was performed according to EC Decision 2002/657. Twenty blank animal feed samples, including milk replacers and wet and dry feed samples, were spiked with 17ß-estradiol (E2ß) at 5ngg ¿1 , 17¿-ethynylestradiol (EE2) at 5ngg ¿1 , diethylstilbestrol (DES) at 10ngg ¿1 , zearalenone at 1.25µgg ¿1 or equol at 200µgg ¿1 . All of these blank and low estrogen spiked feed samples fulfilled the CC¿ and CCß criterions, meaning that all 20 blank feed samples gave a signal below the determined decision limit CC¿ and were thus classified as compliant, and at least 19 out of the 20 spiked samples gave a signal above this CC¿ (ß=5%) and were thus classified as suspect. The method was specific and estrogens in feed were stable for up to 98 days. In this study we also present long-term performance data and several examples of estrogens found in the routine screening of animal feed. This is the first successful example of a developed, validated and applied bioassay for the screening of hormonal substances in feed
Original languageEnglish
Pages (from-to)556-568
JournalFood Additives and Contaminants
Volume23
Issue number6
DOIs
Publication statusPublished - 2006

Keywords

  • green fluorescent protein
  • flight mass-spectrometry
  • in-vitro
  • liquid-chromatography
  • recombinant assay
  • surface waters
  • european eels
  • calf urine
  • expression
  • cells

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