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Quantification of PCDD/Fs and dioxin-like PCBs in small amounts of human serum using the sensitive H1L7.5c1 mouse hepatoma cell line: optimization and analysis of human serum samples from adolescents of the Flemish human biomonitoring program FLEHS II |
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| Authors: | Croes K Van Langenhove K Den Hond E Bruckers L Colles A Koppen G Loots I Nelen V Schoeters G Nawrot T Van Larebeke N Denison M S Vandermarken T Elskens M Baeyens W |
| Institution: | a Vrije Universiteit Brussel, Department of Analytical and Environmental Chemistry (ANCH), Pleinlaan 2, 1050 Brussels, Belgium b Flemish Institute for Technological Research (Vito), Environmental Health and Risk, Boeretang 200, 2400 Mol, Belgium c Interuniversity Institute for Biostatistics and Statistical Bioinformatics, Hasselt University, Belgium d University of Antwerp, Political and Social Sciences, Antwerp, Belgium e Provincial Institute for Hygiene, Antwerp, Belgium f Occupational & Environmental Medicine, Unit of Lung Toxicology, K.U. Leuven, Belgium g Centre for Environmental Sciences, Hasselt University, Diepenbeek Belgium h Ghent University Hospital, Study Centre for Carcinogenesis and Primary Prevention of Cancer, Ghent, Belgium i Department of Environmental Toxicology, University of California, Davis, CA 95616, USA |
| Abstract: | Since the CALUX (Chemically Activated LUciferase gene eXpression) bioassay is a fast and inexpensive tool for the throughput analysis of dioxin-like compounds in a large number of samples and requires only small sample volumes, the use of this technique in human biomonitoring programs provides a good alternative to GC-HRMS. In this study, a method for the separate analysis of PCDD/Fs and dioxin-like PCBs (dl-PCBs) in human serum with the new sensitive H1L7.5c1 mouse hepatoma cell line was optimized.Sample dilution factors of 5 and 2.4 were selected for routine analysis of respectively the PCDD/Fs and dl-PCBs. The validation studies showed that repeatability and within-lab reproducibility for the quality control (QC) standard were within the in-house criteria. A long-term within-lab reproducibility of 25% for the PCDD/F fraction and 41% for the dl-PCB fraction for the analysis of pooled serum samples, expressed as pg BEQ/g fat, was determined. CALUX recoveries of the spiked procedural blanks were within the acceptable in-house limits of 80-120% for both fractions and the LOQ was 30.3 pg BEQ/g fat for the PCDD/Fs and 14.5 pg BEQ/g fat for the dl-PCBs. The GC-HRMS recovery of a C13-spiked pooled serum sample was between 60 and 90% for all PCDD/F congeners and between 67 and 82% for the non-ortho PCBs. An adequate separation between both fractions was found. The CALUX/GC-HRMS ratio for a pooled serum sample was respectively 2.0 and 1.4 for the PCDD/Fs and the dl-PCBs, indicating the presence of additional AhR active compounds. As expected, a correlation was found between human serum samples analyzed with both the new H1L7.5c1 cell line and the more established H1L6.1c3 cell line. The geometric mean CALUX-BEQ values, reported for the adolescents of the second Flemish Environment and Health Study (FLEHS II) recruited in 2009-2010, were 108 (95% CI: 101-114) pg CALUX-BEQ/g fat for the PCDD/Fs and 32.1 (30.1-34.2) pg CALUX-BEQ/g fat for the dioxin-like PCBs. |
| Keywords: | PCDD/Fs Dioxin-like PCBs CALUX Human serum Biomonitoring FLEHS II |
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