QuECHERS as perspective extraction technique for isolation of steroid estrogens from sediments in
                              environmentally relevant concentrations


 Jan Sadílek ^a,^*, Petra Spálovská ^a, Milada Vávrová ^b, Blahoslav Maršálek ^c, Zdeněk Šimek ^a


^a Research Centre for Toxic Compounds in the Environment, Faculty of Science, Masaryk University,
                            Kamenice 753/5, Brno 625 00, Czech Republic

    ^b Institute of Chemistry and Technology of Environmental Protection, Faculty of Chemistry,
               University of Technology, Purkyňova 118, 612 00 Brno, Czech Republic

    ^c  Department of Experimental Phycology and Ecotoxicology, Institute of Botany, Academy of
            Sciences of the Czech republic, Lidická 25/27, Brno 602 00, Czech republic


The presence of endocrine disrupting compounds including steroid estrogens in water environment is
worldwide problem. These compounds are released from different sources such as wastewaters from
household through ineffective wastewater treatment plant also as result of runoff from agricultural
areas after animal manure applications. Steroid estrogens cause a negative effect on growth,
development and reproduction of aquatic organisms at very low concentrations.  Due to higher
octanol-water partition coefficient they tend to sorption and accumulation to solid matrices such
as sediments. For a study of estrogens behavior in water-sediment system the effective extraction
technique is needed. For this reason the four extraction technique: Soxhlet warm extraction (SWE),
Accelerated solvent extraction (ASE), Microwave assisted extraction (MAE) and Quick Easy Cheap
Effective Rugged Safe (QuCHERS) were compared using different extraction conditions. Estron (E1),
17α – estradiol (αE2), 17β- estradiol (βE2), estriol (E3), 17α-ethinilestradiol (EE2) and
17α-ethinilestradiol 2,4,16,16-d4 (EE2d4) were extracted from sediment sample at very low initial
spike concentrations (1ng/g). Wider range of extraction recoveries (40-120 %) was considered as
acceptable in those cases. Simple, fast and inexpensive original QuECHERS with PSA clean up
provided the best extraction recoveries for all tested estrogens (53-84 %). Also MAE achieved the
satisfactory extraction efficiency using DCM:acetone 3:1 as extraction solvent (50-70 %) but only
for less polar estrogens. For estriol the more polar extraction solvents (acetone or methanol) must
be used. More time consuming extraction technique ASE provided lower extraction recovery or worse
repeatability in almost all solvent-temperature combinations in comparison with previous
extraction. The worst results were obtained by most time consuming technique SWE that is totally
unsuitable for this purpose.

Deuterated 17β-estradiol was firstly used in SWE and ASE as internal standard for all tested
estrogens but this compound caused an overestimation of E3. For this reason deuterated
16α-hydroxy-17β-estradiol was selected as preferable internal standard for E3.


Acknowledgment: Reserch was funded by GACR GA13-20357S