Evaluation of Analytical Chemical Methods for Detection of Estrogens in the Environment

Preface

Effluents from municipal and industrial wastewater treatment plants, and agricultural run-off and drainage add numerous exogenous compounds to the aquatic system. Recent studies in a number of countries have shown that the aquatic environment can possess estrogenic activity capable of influencing the fauna. Research has shown that the main substances causing these effects are the natural compounds estrone (E1), 17ß-estradiol (E2) and estriol (E3) and the synthetic estrogen, 17α-ethinylestradiol (EE2). A few other steroid estrogens may also be reason for concern. The primary source of these substances in the environment has hitherto been attributed to human release through sewage treatment however, the question of whether waste from farm animals (cattle and pigs) (17 α-estradiol (E2-17α) is a significant source for the observed effects remains unanswered.

Answering this question requires finding simple, sensitive and specific chemical analytical methods for analysis and screening of samples from wastewater and surface waters for their estrogenic activity. This report evaluates the existing chemical methods, i.e., GC-MS, GC-MS-MS, LC-MS, LC-MS-MS, for determination of estrogenic activity in various environmental matrices. Furthermore, the use of immunochemical methods has been assessed and compared to the chemical analytical methods. The existing knowledge concerning the potential and limitations of these methods is described with the aim of detailing the limitations and draw-backs of available methods for monitoring wastewater and surface water. Furthermore, chemical analytical methods for monitoring agricultural drain water and animal manure slurry have been assessed. A parallel report has been produced assessing existing simple, sensitive and specific in vitro tests for rapid screening of samples from wastewater and surface waters for their estrogenic activity. This report evaluates of the existing in vitro methods for determination of estrogenic activity in various environmental matrices.

This report was written during July and August 2003 to address a scarcity of available information in the published literature on chemical analysis of steroid estrogens in environmental samples. The work was followed by a steering group with the following members:

Inge Vibeke Hansen, Miljøstyrelsen (chairman)
Jørgen Larsen, Miljøstyrelsen
Line Hollesen, Miljøstyrelsen
Nis Hansen, Eurofins
Flemming Ingerslev, Bent Halling-Sørensen Aps
Bent Halling-Sørensen, Bent Halling-Sørensen Aps

The report was written by Flemming Ingerslev and Bent Halling-Sørensen, The Danish University of Pharmaceutical Sciences. Henrik Rasmus Andersen and Kristine Andersen Krogh, The Danish University of Pharmaceutical Sciences has commented on draft versions of the report. This document has been reviewed for English grammar by James Carter of Arroyo Engineering Consultants, Inc., Las Vegas, USA.