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- Analysis of 17- β -estradiol and 17- α -ethinylestradiol in biological and environmental matrices — A reviewPublication . Barreiros, Luisa; Queiroz, Joana F.; Magalhães, Luís M.; Silva, Adrián M.T.; Segundo, Marcela A.The estrogens 17-β-estradiol (E2) and 17-α-ethinylestradiol (EE2) are reported as highly endocrine-disrupting agents, being recently included in an EU watch list regarding emerging aquatic pollutants. Therefore, the monitoring of these chemicals in the different environmental compartments assumes great importance. Moreover, due to the possible adverse effects on living beings, their occurrence on animal tissues and fluids must also be addressed. In recent years, a significant number of studies have described and proposed different analytical methodologies to detect and/or quantify E2 and EE2 mostly in environmental aqueous samples, including sludge and sediments and also in biological matrices such as plasma and tissues. Taking into account the complexity of real matrices and that both estrogens are generally present at trace levels, the development of accurate and reliable techniques for their determination can be quite a challenge. The present review aims at describing the main characteristics of the analytical methods recently used for E2 and EE2 determination in environmental and biological samples. The steps for sample preparation such as analytes extraction, preconcentration and clean-up are discussed and the instrumental based analytical techniques are compared. Furthermore, the application of biological tools to determine the total estrogenicity of environmental samples, as well as their potential combination with instrumental analyses, is highlighted.