Analytical methodologies for determining the occurrence of endocrine disrupting chemicals in sewage treatment plants and natural waters

Abstract In this study, a method for assessing the occurrence of trace amounts of 12 representative estrogenic compounds in sewage and surface waters was developed. The selected substances were the phytoestrogens daidzein, genistein and biochanin A, the alkylphenols bisphenol A and 4-nonylphenol, the natural hormones 17β-estradiol, estrone, estriol and the synthetic hormone 17α-ethynylestradiol and the mycoestrogens zearalenone and two of its metabolites (α-zearalanol and β-zearalanol). The procedure consists in solid phase extraction (SPE) performed with OASIS cartridges followed by liquid chromatography coupled with tandem mass spectrometry (LC–MS–MS). Recoveries were all above 80% for each analyzed aqueous matrices. The developed method was applied to verify the occurrence of endocrine disrupters in environmental samples of sewage influents and effluents of an Italian STP. Phytoestrogens were present in effluents at concentrations ranging from 3 to 83 ng/l, whereas the levels detected for alkylphenols were in the range 13–36 ng/l for bisphenol A and up to 1 μg/l for nonylphenol. Estrogens and zeranols were determined at levels below 30 ng/l. Analysis of a river (Tiber) receiving effluent waters revealed high quantities of bisphenol A (15–29 ng/l) and nonylphenol (up to 1.2 μg/l), whereas the presence of all the other compounds were at levels of few ng/l.

[1]  C. Iden,et al.  Determination of steroid estrogens in wastewater by immunoaffinity extraction coupled with HPLC-electrospray-MS. , 2001, Analytical chemistry.

[2]  H. Greenfield,et al.  Development and validation of an isocratic high-performance liquid chromatographic method for quantitative determination of phytoestrogens in soya bean , 1998 .

[3]  D. Barceló,et al.  Determination of chlorobenzidines in industrial effluent by solid-phase extraction and liquid chromatography with electrochemical and mass spectrometric detection , 1999 .

[4]  Toshitsugu Tanaka,et al.  Co-contamination of mycotoxins in corn harvested in Haimen, a high risk area of primary liver cancer in China , 1995 .

[5]  J. Kellis,et al.  Inhibition of human estrogen synthetase (aromatase) by flavones. , 1984, Science.

[6]  Shane A. Snyder,et al.  Analytical Methods for Detection of Selected Estrogenic Compounds in Aqueous Mixtures , 1999 .

[7]  P. Scott,et al.  Risk assessment of the mycotoxin zearalenone. , 1987, Regulatory toxicology and pharmacology : RTP.

[8]  W. Mazur,et al.  Isoflavonoids and Lignans in Legumes: Nutritional and Health Aspects in Humans 1 1 The method develo , 1998 .

[9]  M. Comber,et al.  The effects of nonylphenol on Daphnia magna , 1993 .

[10]  K. Ballschmiter,et al.  Determination of endocrine-disrupting phenolic compounds and estrogens in surface and drinking water by HRGC-(NCI)-MS in the picogram per liter range. , 2001, Environmental science & technology.

[11]  R. Samperi,et al.  Trace Analysis of Sulfonylurea Herbicides in Water: Extraction and Purification by a Carbograph 4 Cartridge, Followed by Liquid Chromatography with UV Detection, and Confirmatory Analysis by an Electrospray/Mass Detector , 1997 .

[12]  K. Schwadorf,et al.  Natural occurrence of Fusarium toxins in oats harvested during five years in an area of southwest Germany. , 1998, Food additives and contaminants.

[13]  K. Fytianos,et al.  Determination of non-ionic surfactants (polyethoxylated-nonylphenols) by HPLC in waste waters , 1997 .

[14]  S X Peng,et al.  Fully automated 96-well liquid-liquid extraction for analysis of biological samples by liquid chromatography with tandem mass spectrometry. , 2001, Analytical chemistry.

[15]  D. Barceló,et al.  Use of solid-phase extraction in various of its modalities for sample preparation in the determination of estrogens and progestogens in sediment and water. , 2001, Journal of chromatography. A.

[16]  J. Sumpter,et al.  Estrogenic Effects of Effluents from Sewage Treatment Works , 1994 .

[17]  M. Depledge,et al.  Ecological Significance of Endocrine Disruption in Marine Invertebrates , 1999 .

[18]  A. Laganà,et al.  Development of an analytical system for the simultaneous determination of anabolic macrocyclic lactones in aquatic environmental samples. , 2001, Rapid communications in mass spectrometry : RCM.

[19]  V A Baker,et al.  Endocrine disrupters--testing strategies to assess human hazard. , 2001, Toxicology in vitro : an international journal published in association with BIBRA.

[20]  P Zöllner,et al.  Determination of zearalenone in grains by high-performance liquid chromatography-tandem mass spectrometry after solid-phase extraction with RP-18 columns or immunoaffinity columns. , 1999, Journal of chromatography. A.

[21]  L. R. Harris,et al.  Bisphenol A concentrations in receiving waters near US manufacturing and processing facilities. , 2000, Chemosphere.

[22]  P. Zöllner,et al.  Determination of zeranol, taleranol, zearalenone, α- and β-zearalenol in urine and tissue by high-performance liquid chromatography-tandem mass spectrometry , 2000 .

[23]  D. Barceló,et al.  Estrogenicity determination in sewage treatment plants and surface waters from the Catalonian area (NE Spain) , 2000 .

[24]  M. Grasserbauer,et al.  Comparison of different sorbent materials for on-line solid-phase extraction with liquid chromatography-atmospheric pressure chemical ionization mass spectrometry of phenols. , 2000, Journal of chromatography. A.

[25]  M. Ronis,et al.  An LC-MS method to determine concentrations of isoflavones and their sulfate and glucuronide conjugates in urine. , 1999, Clinica chimica acta; international journal of clinical chemistry.

[26]  R. Giese,et al.  General method for determining ethylene oxide and related N7-guanine DNA adducts by gas chromatography-electron capture mass spectrometry. , 1995, Journal of chromatography. A.

[27]  M. Servos,et al.  Behavior and occurrence of estrogens in municipal sewage treatment plants--I. Investigations in Germany, Canada and Brazil. , 1999, The Science of the total environment.

[28]  A. Johnson,et al.  Estimating steroid oestrogen inputs into activated sludge treatment works and observations on their removal from the effluent. , 2000, The Science of the total environment.

[29]  M. Waldock,et al.  Concentrations of alkylphenols in rivers and estuaries in England and Wales , 1995 .

[30]  M. Pettersson,et al.  ETHINYLOESTRADIOL - AN UNDESIRED FISH CONTRACEPTIVE , 1999 .

[31]  A. Scott,et al.  Survey of estrogenic activity in United Kingdom estuarine and coastal waters and its effects on gonadal development of the flounder Platichthys flesus , 1999 .

[32]  J. Sumpter,et al.  Removal of endocrine-disrupting chemicals in activated sludge treatment works. , 2001, Environmental science & technology.

[33]  Motoyama,et al.  Direct determination of bisphenol A and nonylphenol in river water by column-switching semi-microcolumn liquid chromatography/electrospray mass spectrometry , 1999, Rapid communications in mass spectrometry : RCM.

[34]  Edwin J. Routledge,et al.  Identification of Estrogenic Chemicals in STW Effluent. 1. Chemical Fractionation and in Vitro Biological Screening , 1998 .