Sample preparation for combined chemical analysis and in vitro bioassay application in water quality assessment.
The combination of in vitro bioassays and chemical screening can provide a powerful toolbox to determine biologically relevant compounds in water extracts. In this study, a sample preparation method is evaluated for the suitability for both chemical analysis and in vitro bioassays. A set of 39 chemicals were spiked to surface water, which were extracted using Oasis MCX cartridges. The extracts were chemically analyzed by liquid chromatography linear ion trap Orbitrap analysis and recoveries appeared to be on average 61% Compounds with logK(ow) values in the range between 0 and 4 are recovered well using this method. In a next step, the same extracts were tested for genotoxic activity using the Comet assay and Ames fluctuation test and for specific endocrine receptor activation using a panel of CALUX assays, for estrogenic (ER), androgenic (AR), glucocorticoid (GR), progestagenic (PR), and thyroidogenic (TR) agonistic activities. The results of the genotoxicity assays indicated that spiked genotoxic compounds were preserved during sample preparation. The measured responses of the GR CALUX and ER CALUX assays were similar to the predicted responses. The measured responses in the AR CALUX and PR CALUX assays were much lower than expected from the analytical concentration, probably due to antagonistic effects of some spiked compounds. Overall, the presented sample preparation method seems to be suitable for both chemical analysis and specific in vitro bioassay applications.
KWR Watercycle Research Institute, P.O. Box 1072, 3430 BB Nieuwegein, The Netherlands. Electronic address: annemieke.kolkman@kwrwater.nl.
Annemieke
A
Kolkman
Merijn
M
Schriks
Walter
W
Brand
Patrick S
PS
Bäuerlein
Margaretha M E
MM
van der Kooi
René H
RH
van Doorn
Erik
E
Emke
Astrid A
AA
Reus
Sander C
SC
van der Linden
Pim
P
de Voogt
Minne B
MB
Heringa
D000818:Animals; D001681:Biological Assay / Q000379:methods*; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid; D020552:Comet Assay; D004249:DNA Damage; D060766:Drinking Water / Q000032:analysis; D052244:Endocrine Disruptors / Q000633:toxicity; D006801:Humans; D013058:Mass Spectrometry; D009152:Mutagenicity Tests; D051381:Rats; D013048:Specimen Handling; D060753:Water Quality / Q000592:standards*; D014881:Water Supply / Q000032:analysis*
D060766:Drinking Water; D052244:Endocrine Disruptors
Chemical screening; High resolution mass spectrometry; In vitro bioassays; Sample preparation; Solid phase extraction