Quantitative Analysis of Natural and Synthetic Estrogens in Surface and Final Effluent Waters at Low ppq Levels Using UPLC-MS/MS

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Euan Ross, Benjamin Wuyts, and Angela Boag
Waters and Scottish Water
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Application Notes
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Application Notes
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Estrogens are routinely used either as contraceptive medicines or in hormone replacement therapy and they can enter aquatic environments via the discharge of final effluent waters. Estrogens are believed to have a negative effect on aquatic environments by disrupting the hormonal systems of fish. For the European Union, EU Directive 2013/39/EU includes 15 additional priority substances to the water framework Directive 2000/60/EC. In this update, 17α-ethinylestradiol and 17β-estradiol were not included in the priority substance list, but instead added to a watch list in order to gather further data regarding the  presence of these compounds in aquatic environments, and the risks they may pose.

This application note summarizes a method for the analysis of estrone, 17α-ethinylestradiol and 17β-estradiol in surface and final effluent waters. To achieve the challenging EU LLOQ parts per quadrillion (ppq) levels required for these compounds, a combination of off-line solid phase extraction (SPE) clean-up and pre-concentration, combined with a large volume injection and triple quadrupole mass spectrometry were utilized.

Application benefits include:

  • High sensitivity to achieve EU target LLOQs.
  • Baseline separation and good peak shapes for target analytes.
  • Suitable linearity and repeatability data in surface water and final effluent.
  • Acceptable SPE spiked recoveries and repeatability.
  • Quantitation of detected residues in final effluent using standard addition in the absence of isotopically labeled internal standard.

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