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)

K. Croes, K. Van Langenhove, E. Den Hond, L. Bruckers, A. Colles, G. Koppen, I. Loots, V. Nelen, G. Schoeters, T. Nawrot, N. Van Larebeke, M. S. Denison, T. Vandermarken, M. Elskens, W. Baeyens

Research output: Contribution to journalArticle

9 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)2484-2491
Number of pages8
JournalTalanta
Volume85
Issue number5
DOIs
StatePublished - Oct 15 2011

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Keywords

  • Biomonitoring
  • CALUX
  • Dioxin-like PCBs
  • FLEHS II
  • Human serum
  • PCDD/Fs

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Croes, K., Van Langenhove, K., Hond, E. D., 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. (2011). 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). Talanta, 85(5), 2484-2491. https://doi.org/10.1016/j.talanta.2011.07.103