Coupling Sample Preparation with Effect-Directed Analysis of Estrogenic Activity - Proposal for a New Rapid Screening Concept for Water Samples

; (). Coupling Sample Preparation with Effect-Directed Analysis of Estrogenic Activity - Proposal for a New Rapid Screening Concept for Water Samples. Journal of Planar Chromatography, 26, 5. 402-408. Peer reviewed.

A new concept coupling sample preparation (using thin-layer chromatography) with effect-directed analysis (TLC–EDA) is proposed and demonstrated, using the planar-YES (a variation of the yeast estrogen screen, YES) as example. The concept allows performing on one single TLC layer all steps including sample preparation, sample separation and the detection of bio-effective substances and also offers a potential link to high-end chemical analytics. Estrogen standards were separated by TLC, distinguished and detected by a biological test system. Dose–response curves were produced for 17β-estradiol (E2) and 17α-ethinylestradiol (EE2). In a field trial, bioactive compounds were extracted from a recirculating fish culture and from a wastewater treatment plant and tested. Low concentrations of a substance likely to be E2 were found in both samples. The level of detection for E2 and EE2 at the current stage of development was 0.5 pg per band (5 mm × 1 mm). The time demand for incubation, compared to the standard YES procedure of 1996, was reduced from >20 hours to 5 hours. If the level of detection can be lowered a little more, by less than one order of magnitude, a rapid screening of native aqueous samples on estrogenic activity is realistic. This will open a wide range of different fields of application.