A new bioautographic screening method for the detection of estrogenic compounds

摘要

The yeast estrogen screen was introduced as biological detection method for high performance thin layer chromatography. Yeast cells were grown directly on HPTLC plates, where in the presence of estrogenic substances the production of the enzyme beta-galactosidase was induced. For the natural ligand 17 beta-estradiol, sensitivity could be improved by a factor of 20 using the fluorogenic substrate 4-methylumbelliferyl beta-D-galactopyranoside instead of the chromogenic substrate chlorophenol red -beta-D-galactopyranoside. The fluorescence intensity of "estrogenic" zones was measured using a commercial TLC-Scanner. A nearly full dose-response curve was obtained for 17 beta-estradiol masses between 2.75 and 550 pg, or concentrations of 2.75 to 550 mug L-1.