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Title
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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 Environment and Health Study (FLEHS II)
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Author
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Abstract
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| 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 GCHRMS. 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 80120% 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 GCHRMS 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/GCHRMS 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 20092010, were 108 (95% CI: 101114) pg CALUX-BEQ/g fat for the PCDD/Fs and 32.1 (30.134.2) pg CALUX-BEQ/g fat for the dioxin-like PCBs. |
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Language
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English
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Source (journal)
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Talanta : the international journal of pure and applied analytical chemistry. - Oxford, 1958, currens
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Publication
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Oxford
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Pergamon
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2011
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ISSN
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0039-9140
[print]
1873-3573
[online]
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DOI
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10.1016/J.TALANTA.2011.07.103
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Volume/pages
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85
:5
(2011)
, p. 2484-2491
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ISI
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000296115600033
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Full text (Publisher's DOI)
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Full text (publisher's version - intranet only)
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