Determination of Estrogens in Sludge and Sediments by Liquid Extraction and GC/MS/MS Thomas A. Ternes,* ,² Henrik Andersen, ‡ Daniel Gilberg, § and Matthias Bonerz ² ESWE-Institute for Water Research and Water Technology, D-65201 Wiesbaden, Soehnleinstrasse 158, Germany, The Royal Danish School of Pharmacy, Institute of Analytical and Pharmaceutical Chemistry, Section for Environmental Chemistry, Universitetsparken 2, DK-2100 Copenhagen, Denmark, and Ecotoxicology GmbH, Bo ¨ ttgerstrasse 2-14, D-60437 Flo ¨ rsheim am Main, Germany Two methods have been developed that enable the deter- mination of estrogens down to 2 ng/ g in digested and activated sludge from domestic sewage treatment plants (STPs) and down to 0 .2 ng/ g in freshwater sediments. The method for sludge analysis consists of solvent extrac- tion; a gel permeation chromatography (GPC) cleanup step, a 1 g silica gel column; and finally, detection by GC- ion trap MS/ MS of the silylated estrogens with MSTFA. For sediments, the solvent extraction was successively followed by silica gel cleanup, solid phase enrichment (SPE), and a HPLC cleanup before derivatization and GC/ MS/ MS detection. Mean recoveries of the estrogens mainly exceeded 7 0 % in sludge and 9 0 % in sediments. In activated and digested sewage sludge, estrone and 17 - estradiol were detected up to 3 7 ng/ g and 4 9 ng/ g, respectively, and 1 7 r-ethinylestradiol up to 1 7 ng/ g. The occurrence of estrogens in digested sludge indicates that estrogens can be persistent during sludge digestion. In river sediments, estrone and 1 7 -estradiol were detected up to 2 ng/ g (estrone), and the contraceptive 1 7 r- ethinylestradiol was found with a maximum of 0 .9 ng/ g. Mestranol, a prodrug for 1 7 r-ethinylestradiol, was not detected either in sludge or in sediments. Endocrine-disrupting effects in the aquatic environment, such as the feminization of male fish can probably be attributed to the presence of estrogens in river water. 1-3 Many other substances, such as alkylphenol, phthalic esters, PCBs, and phytoestrogens are suspected to influence the hormonal system, as well. 4 Estrogens are extremely potent compounds and estrogenic effects have been observed in laboratory studies down to 1 ng/ L. 3 Worldwide, in municipal sewage treatment plant (STP) discharges and in the receiving waters, predominantly estrone, 17-estradiol, and the contraceptive 17R-ethinylestradiol have been detected. Concentration levels of only a few nanograms per liter or even down to the picograms-per-liter range 2,5-11 have been reported. The log K OW of 3.1-4.7 indicates that estrogens are rather lipophilic and should appreciably adsorb onto sediment and sludge (Table 1). This assumption is underscored by the detection of high concentrations of estrogens in water released by dewatering sewage sludge. 12 Therefore, a potential contamination of soil with estrogens may be caused by the application of digested sludge from municipal STPs onto agricultural fields. Further, it seems likely that estrogens are present in sediments, and because of their extremely high estrogenic potency, the possible threat to sediment biota cannot be ruled out. Analytical methods for the determination of estrogens have been described in the literature for aqueous matrixes, such as wastewater and river water. The methods published for the determination of estrogens in water are frequently based on SPE, silylation and detection by GC/ MS or GC-ion trap MS/ MS. The LOQs are in the lower nanograms-per-liter range for such methods. 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