View More View Less
  • 1 University of Offenburg, Badstrasse 24, D-77652 Offenburg, Germany
Restricted access

We present a two-dimensional (2D) planar chromatographic separation of estrogenic active compounds on RP-18 W (Merck, 1.14296) phase. A mixture of 8 substances was separated using a solvent mix consisting of hexane, ethyl acetate, acetone (55:15:10, v/v) in the first direction and of acetone and water (15:10, v/v) in the second direction. Separation was performed on an RP-18 W plate over a distance of 70 mm. This 2D-separation method can be used to quantify 17α-ethinylestradiol (EE2) in an effect-directed analysis, using the yeast strain Saccharomyces cerevisiae BJ3505. The test strain (according to McDonnell) contains the estrogen receptor. Its activation by estrogen active compounds is measured by inducing the reporter gene lacZ which encodes the enzyme β-galactosidase. This enzyme activity is determined on plate by using the fluorescent substrate MUG (4-methylumbelliferyl-β-d-galactopyranoside).

  • [1]

    B. Milz , B. Spangenberg, J. Liq. Chromatogr. Relat. Technol. 36 (2013) 23782386.

  • [2]

    B. Milz , B. Spangenberg, J. Liq. Chromatogr. Relat. Technol. 37 (2014) 28722881.

  • [3]

    A. Tiehm , A. Huber, S. Mala, C. Stoll, Evaluierung und Anwendung des Yeast-Estrogen-Screen (YES)-Assays für den summarischen Nachweis des Vorkommens und Abbaus von endokrinen Substanzen, DVGW, Karlsruhe, 2014.

    • Search Google Scholar
    • Export Citation
  • [4]

    E. J. Routledge , J. P. Sumpter, Environ. Toxicol. Chem. 15 (1996) 241248.

  • [5]

    D. P. McDonnell , Z. Nawaz, C. Densmore, N. L. Weigel, T. A. Pham, J. H. Clark, B. W. O’Malley, J. Steroid Biochem. Mol. Biol. 39 (1991) 291297.

    • Search Google Scholar
    • Export Citation
  • [6]

    M. B. Müller , C. Dausend, C. Weins, F. H. Frimmel, Chromatographia 60 (2004) 207211.

  • [7]

    D. Spira , G. Reifferscheid, S. Buchinger, J. Planar Chromatogr. 26 (2013) 395401.

  • [8]

    A. Schönborn , A. Grimmer, J. Planar Chromatogr. 26 (2013) 402408.

  • [9]

    I. Klingelhöfer , G. Morlock, J. Chromatogr. A 1360 (2014) 288295.

  • [10]

    International Standard ISO 7027 for values between 40 FAU and 4000 FAU.

  • [11]

    B. Spangenberg , C. F. Poole, C. Weins, Quantitative Thin Layer Chromatography, A Practical Survey, Springer, Berlin, 2010.

  • [12]

    S. Ilic , M. Natic, D. Dabic, D. Milojkovic-Opsenica, Ž. Tešic, J. Planar Chromatogr. 24 (2011) 9398.

  • [13]

    B. Spangenberg , J. Planar Chromatogr. 28 (2015) 139143.

  • [14]

    R. L. Gwarda , T. H. Dzido, J. Chromatogr. A 1312 (2013) 152154.

  • [15]

    C. F. Poole , Instrumental Thin-Layer Chromatography, Elsevier, Amsterdam, 2015.

  • [16]

    H. Jork , W. Funk, W. Fischer, H. Wimmer, Thin-Layer Chromatography, Reagents and Detection Methods, VCH, Weinheim, 1994.

  • [17]

    I. Klingelhoefer , G. Morlock, Bioassays on modified TLC/ HPTLC plates, European Patent Application EP2919004, 2015.

  • [18]

    W. Funk , V. Dammann, G. Donnevert, Quality Assurance in Analytical Chemistry, 2nd edn., Wiley-VCH, Weinheim, 2007.

Monthly Content Usage

Abstract Views Full Text Views PDF Downloads
Jun 2020 0 1 2
Jul 2020 11 1 0
Aug 2020 1 0 0
Sep 2020 10 0 0
Oct 2020 5 0 0
Nov 2020 2 0 0
Dec 2020 0 0 0