Analytical and Bioanalytical Chemistry

, Volume 408, Issue 24, pp 6833–6844 | Cite as

Application of microwave-assisted extraction and ultra-high performance liquid chromatography–tandem mass spectrometry for the analysis of sex hormones and corticosteroids in sewage sludge samples

  • Rayco Guedes-Alonso
  • Sergio Santana-Viera
  • Sarah Montesdeoca-Esponda
  • Cristina Afonso-Olivares
  • Zoraida Sosa-Ferrera
  • José Juan Santana-RodríguezEmail author
Research Paper


Hormonal compounds are a concern to the international community because they can affect the aquatic biota and are therefore considered to be endocrine-disrupting compounds. These compounds have lipophilic properties, so they tend to accumulate in solid matrices, such as sewage sludge. This work presents the optimization of a microwave-assisted extraction process combined with ultra-high performance liquid chromatography–tandem mass spectrometry for the determination of 15 hormonal compounds in sludge samples. The proposed method has relative standard deviations below 23 %, good recoveries (over 71 %) for all compounds, detection limits that ranged from 1.1 to 7.9 ng g−1 and quantification limits which ranged from 3.7 to 26.3 ng g−1. The method was used to analyse sludge samples from four different wastewater treatment plants of Gran Canaria (Spain) with different wastewater treatments. 17β-estradiol, 17α-ethynylestradiol, norgestrel and cortisone were detected in sludge samples at concentrations that ranged from 17.3 to 1.44 × 103 ng g−1. The developed method permits the use of small quantities of sample and organic solvents, presents short extractions times and is the first one based on microwave-assisted extraction for the analysis of both sex hormones and corticosteroids.


Mass spectrometry Microwave-assisted extraction Sludge Steroid hormones Ultra-high performance liquid chromatography 



This work was supported by funds provided by the Spanish Ministry of Economy and Competitiveness, Research Project CTM2015-66095-C2-1-R. Rayco Guedes-Alonso thanks the University of Las Palmas de Gran Canaria (Spain) for his Ph.D. student grant. The authors would also like to thank Emalsa S.A., Acciona S.A. and Consejo Insular de Aguas de Gran Canaria for allowing the sampling of sludge at the different wastewater treatment plants.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.


  1. 1.
    Krause DN, Duckles SP, Pelligrino DA. Influence of sex steroid hormones on cerebrovascular function. J Appl Physiol. 2006;101:1252–61. doi: 10.1152/japplphysiol.01095.2005.CrossRefGoogle Scholar
  2. 2.
    Mayes JS, Watson GH. Direct effects of sex steroid hormones on adipose tissues and obesity. Obes Rev. 2004;5:197–216. doi: 10.1111/j.1467-789X.2004.00152.x.CrossRefGoogle Scholar
  3. 3.
    Kugathas S, Williams RJ, Sumpter JP. Prediction of environmental concentrations of glucocorticoids: the River Thames, UK, as an example. Environ Int. 2012;40:15–23. doi: 10.1016/j.envint.2011.11.007.CrossRefGoogle Scholar
  4. 4.
    Besse J-P, Garric J. Progestagens for human use, exposure and hazard assessment for the aquatic environment. Environ Pollut. 2009;157:3485–94. doi: 10.1016/j.envpol.2009.06.012.CrossRefGoogle Scholar
  5. 5.
    Hoffman JR, Kraemer WJ, Bhasin S, Storer T, Ratamess NA, Haff GG, et al. Position stand on androgen and human growth hormone use. J Strength Cond Res. 2009;23:S1–59. doi: 10.1519/JSC.0b013e31819df2e6.CrossRefGoogle Scholar
  6. 6.
    Chang H, Hu J, Shao B. Occurrence of natural and synthetic glucocorticoids in sewage treatment plants and receiving river waters. Environ Sci Technol. 2007;41:3462–8. doi: 10.1021/es062746o.CrossRefGoogle Scholar
  7. 7.
    Chang H, Wan Y, Wu S, Fan Z, Hu J. Occurrence of androgens and progestogens in wastewater treatment plants and receiving river waters: comparison to estrogens. Water Res. 2011;45:732–40. doi: 10.1016/j.watres.2010.08.046.CrossRefGoogle Scholar
  8. 8.
    Servos MR, Bennie DT, Burnison BK, Jurkovic A, McInnis R, Neheli T, et al. Distribution of estrogens, 17β-estradiol and estrone, in Canadian municipal wastewater treatment plants. Sci Total Environ. 2005;336:155–70. doi: 10.1016/j.scitotenv.2004.05.025.CrossRefGoogle Scholar
  9. 9.
    Racz L, Goel RK. Fate and removal of estrogens in municipal wastewater. J Environ Monit. 2010;12:58–70. doi: 10.1039/b917298j.CrossRefGoogle Scholar
  10. 10.
    Colman JR, Baldwin D, Johnson LL, Scholz NL. Effects of the synthetic estrogen, 17α-ethinylestradiol, on aggression and courtship behavior in male zebrafish (Danio rerio). Aquat Toxicol. 2009;91:346–54. doi: 10.1016/j.aquatox.2008.12.001.CrossRefGoogle Scholar
  11. 11.
    Guedes-Alonso R, Sosa-Ferrera Z, Santana-Rodriguez JJ. On-line solid phase extractionmethod coupled with UHPLC-MS/MS for the determination of steroid hormone compounds in treated waters from wastewater treatment plants. Anal Methods. 2015. doi: 10.1039/C5AY00807G.Google Scholar
  12. 12.
    Pessoa GP, de Souza NC, Vidal CB, Alves JAC, Firmino PIM, Nascimento RF, et al. Occurrence and removal of estrogens in Brazilian wastewater treatment plants. Sci Total Environ. 2014;490:288–95. doi: 10.1016/j.scitotenv.2014.05.008.CrossRefGoogle Scholar
  13. 13.
    Liu S, Ying G-G, Zhao J-L, Chen F, Yang B, Zhou L-J, et al. Trace analysis of 28 steroids in surface water, wastewater and sludge samples by rapid resolution liquid chromatography–electrospray ionization tandem mass spectrometry. J Chromatogr A. 2011;1218:1367–78. doi: 10.1016/j.chroma.2011.01.014.CrossRefGoogle Scholar
  14. 14.
    Pal A, Gin KY-H, Lin AY-C, Reinhard M. Impacts of emerging organic contaminants on freshwater resources: review of recent occurrences, sources, fate and effects. Sci Total Environ. 2010;408:6062–9. doi: 10.1016/j.scitotenv.2010.09.026.CrossRefGoogle Scholar
  15. 15.
    Jurado A, Vàzquez-Suñé E, Carrera J, de Alda López M, Pujades E, Barceló D. Emerging organic contaminants in groundwater in Spain: A review of sources, recent occurrence and fate in a European context. Sci Total Environ. 2012;440:82–94. doi: 10.1016/j.scitotenv.2012.08.029.CrossRefGoogle Scholar
  16. 16.
    Vega-Morales T, Sosa-Ferrera Z, Santana-Rodríguez JJ. Determination of various estradiol mimicking-compounds in sewage sludge by the combination of microwave-assisted extraction and LC–MS/MS. Talanta. 2011;85:1825–34. doi: 10.1016/j.talanta.2011.07.051.CrossRefGoogle Scholar
  17. 17.
    Martín J, Santos JL, Aparicio I, Alonso E. Multi-residue method for the analysis of pharmaceutical compounds in sewage sludge, compost and sediments by sonication-assisted extraction and LC determination. J Sep Sci. 2010;33:1760–6. doi: 10.1002/jssc.200900873.CrossRefGoogle Scholar
  18. 18.
    Herrero P, Borrull F, Marcé RM, Pocurull E. Pressurised liquid extraction and ultra-high performance liquid chromatography-tandem mass spectrometry to determine endogenous and synthetic glucocorticoids in sewage sludge. Talanta. 2013;103:186–93. doi: 10.1016/j.talanta.2012.10.030.CrossRefGoogle Scholar
  19. 19.
    Snow DD, Damon-Powell T, Onanong S, Cassada DA. Sensitive and simplified analysis of natural and synthetic steroids in water and solids using on-line solid-phase extraction and microwave-assisted solvent extraction coupled to liquid chromatography tandem mass spectrometry atmospheric pressure photoionization. Anal Bioanal Chem. 2013;405:1759–71. doi: 10.1007/s00216-012-6572-8.CrossRefGoogle Scholar
  20. 20.
    Temes TA, Andersen H, Gilberg D, Bonerz M. Determination of estrogens in sludge and sediments by liquid extraction and GC/MS/MS. Anal Chem. 2002;74:3498–504.CrossRefGoogle Scholar
  21. 21.
    Muller M, Rabenoelina F, Balaguer P, Patureau D, Lemenach K, Budzinski H, et al. Chemical and biological analysis of endocrine-disrupting hormones and estrogenic activity in an advanced sewage treatment plant. Environ Toxicol Chem. 2008;27:1649–58. doi: 10.1897/07-519.1.CrossRefGoogle Scholar
  22. 22.
    Gineys N, Giroud B, Vulliet E. Analytical method for the determination of trace levels of steroid hormones and corticosteroids in soil, based on PLE/SPE/LC-MS/MS. Anal Bioanal Chem. 2010;397:2295–302. doi: 10.1007/s00216-010-3787-4.CrossRefGoogle Scholar
  23. 23.
    Wang H, Zhou X, Zhang Y, Chen H, Li G, Xu Y, et al. Dynamic microwave-assisted extraction coupled with salting-out liquid–liquid extraction for determination of steroid hormones in fish tissues. J Agric Food Chem. 2012;60:10343–51. doi: 10.1021/jf303124c.CrossRefGoogle Scholar
  24. 24.
    Vega Morales T, Torres Padrón ME, Sosa Ferrera Z, Santana Rodríguez JJ. Determination of alkylphenol ethoxylates and their degradation products in liquid and solid samples. TrAC Trends Anal Chem. 2009;28:1186–200. doi: 10.1016/j.trac.2009.07.011.CrossRefGoogle Scholar
  25. 25.
    Vieno NM, Tuhkanen T, Kronberg L. Analysis of neutral and basic pharmaceuticals in sewage treatment plants and in recipient rivers using solid phase extraction and liquid chromatography–tandem mass spectrometry detection. J Chromatogr A. 2006;1134:101–11. doi: 10.1016/j.chroma.2006.08.077.CrossRefGoogle Scholar
  26. 26.
    Yu Y, Huang Q, Cui J, Zhang K, Tang C, Peng X. Determination of pharmaceuticals, steroid hormones, and endocrine-disrupting personal care products in sewage sludge by ultra-high-performance liquid chromatography–tandem mass spectrometry. Anal Bioanal Chem. 2011;399:891–902. doi: 10.1007/s00216-010-4295-2.CrossRefGoogle Scholar
  27. 27.
    Ying G-G, Kookana RS. Sorption and degradation of estrogen-like-endocrine disrupting chemicals in soil. Environ Toxicol Chem. 2005;24:2640–5. doi: 10.1897/05-074R.1.CrossRefGoogle Scholar
  28. 28.
    Ren Y-X, Nakano K, Nomura M, Chiba N, Nishimura O. A thermodynamic analysis on adsorption of estrogens in activated sludge process. Water Res. 2007;41:2341–8. doi: 10.1016/j.watres.2007.01.058.CrossRefGoogle Scholar
  29. 29.
    Liu S, Ying G-G, Zhao J-L, Zhou L-J, Yang B, Chen Z-F, et al. Occurrence and fate of androgens, estrogens, glucocorticoids and progestagens in two different types of municipal wastewater treatment plants. J Environ Monit. 2012;14:482–91. doi: 10.1039/C1EM10783F.CrossRefGoogle Scholar
  30. 30.
    Kuster M, de Alda José López M, Barceló D. Analysis and distribution of estrogens and progestogens in sewage sludge, soils and sediments. TrAC Trends Anal Chem. 2004;23:790–8. doi: 10.1016/j.trac.2004.08.007.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Rayco Guedes-Alonso
    • 1
  • Sergio Santana-Viera
    • 1
  • Sarah Montesdeoca-Esponda
    • 1
  • Cristina Afonso-Olivares
    • 1
  • Zoraida Sosa-Ferrera
    • 1
  • José Juan Santana-Rodríguez
    • 1
    Email author
  1. 1.Instituto Universitario de Estudios Ambientales y Recursos Naturales (i-UNAT)Universidad de Las Palmas de Gran CanariaLas Palmas de Gran CanariaSpain

Personalised recommendations