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Bovee, T.F.H.* ; Bor, G.* ; Becue, I.* ; Daamen, F.E.J.* ; van Duursen, M.B.M.* ; Lehmann, S.* ; Vollmer, G.* ; de Maria, R.* ; Fox, J.E.* ; Witters, H.* ; Bernhöft, S. ; Schramm, K.-W. ; Hoogenboom, R.L.A.P.* ; Nielen, M.W.F.*

Inter-laboratory comparison of a yeast bioassay for the determination of estrogenic activity in biological samples.

Anal. Chim. Acta 637, (Sp. Iss. SI), 265-272 (2009)

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An inter-laboratory exercise was performed with a yeast estrogen bioassay, based on the expression of yeast enhanced green fluorescent protein (yEGFP), for the determination of estrogenic activity in extracts of calf urine samples. Urine samples were spiked with 1 and 5 ngmL(-1) 17beta-estradiol and 17alpha-ethynylestradiol, 10 and 50 ngmL(-1) mestranol, and 100 ngmL(-1) testosterone and progesterone. Sample extracts of blank and spiked urine samples were prepared at our laboratory and sent to seven laboratories together with a reagent blank, a DMSO blank, and eight 17beta-estradiol stock solutions in DMSO ranging in concentration from 0 to 545 ngmL(-1). Sample extracts and standards were coded and tested blindly. A decision limit (CCalpha) was determined based on the response of seven blank urine samples. Signals of the negative controls, e.g. urine samples spiked with 100 ngmL(-1) testosterone or progesterone, were all below the determined CCalpha and were thus screened as compliant. Positive controls, i.e. the urine samples spiked at two levels with 17beta-estradiol, 17alpha-ethynylestradiol and mestranol, were almost all screened as suspect, i.e. gave signals above the determined CCalpha. Determined EC(50) values calculated from the 17beta-estradiol dose-response curves obtained by the seven laboratories ranged from 0.59 to 0.95 nM.

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Publication type Article: Journal article

Document type Scientific Article

Corresponding Author

Keywords Bioassay; Estrogens; Transfer validation; Gas chromatography tandem mass spectrometry; tandem mass-spectrometry; green fluorescent protein; liquid-chromatography; anabolic-steroids; recombinant assay; surface waters; beta-agonists; urine; validation; abus

ISSN (print) / ISBN 0003-2670

e-ISSN 1873-4324

Journal Analytica Chimica Acta

Quellenangaben Volume: 637, Issue: 1-2, Pages: 265-272, Supplement: (Sp. Iss. SI)

Publisher Elsevier

Publishing Place Amsterdam

Non-patent literature Publications

Reviewing status Peer reviewed

Institute(s) Institute of Ecological Chemistry (IOEC)