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Dioxin-Like TEQ of women from the Seveso, Italy area by ID-HRGC/HRMS and CALUX

Abstract

Polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and polychlorinated biphenyls (PCBs) are widespread environmental contaminants that exist as complex mixtures and are frequently detected at part-per-trillion (ppt) levels in humans. Using isotope dilution high-resolution gas chromatography/ high-resolution mass spectrometry (HRGC/HRMS), we measure the PCDDs, PCDFs, and PCBs in serum of a population of 78 women residing in an area near Seveso, Italy where a TCDD explosion occurred in 1976 and where furniture is manufactured. The average total dioxin-like toxic equivalents (TEQ) of these women was 25.3 ppt, lipid-adjusted, comparable to other parts of Europe. TCDD levels, however, were higher among the few women who resided in the exposed area in 1976. We examined the possibility of using the CALUX (chemical-activated luciferase gene expression) bioassay to estimate total TEQ in a small volume of plasma from this population. A total of 32 archived plasma specimens were selected for CALUX bioassay, based on the distribution of Total TEQ by HRGC/HRMS. The CALUX bioassay was performed blind to HRGC/HRMS results with 2 ml plasma per replicate analysis. Of 32 samples, 10 were below detection limits in the CALUX bioassay. For the 32 samples, the CALUX-TEQ averaged 25.4 ppt, lipid-adjusted (range: 0–127.6) and was not significantly different from the HRGC/HRMS Total TEQ average of 31.2 ppt, lipid-adjusted (range: 12.7–88.3) (t=0.88, P=0.38), however, the two measures were not significantly correlated (Rs=0.04, P=0.82). More validation of the CALUX bioassay with larger sample volume is needed before application as an exposure measure in large-scale epidemiologic studies of health effects of dioxin-like compounds.

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References

  • Aarts J., Cenjin P., and Blankvoort B., et al. Application of the chemical-activated luciferase expression (CALUX) bioassay for quantification of dioxin-like compounds in small samples of human milk and blood plasma. Organohalogen Compds 1996: 27: 285–290.

    CAS  Google Scholar 

  • Akins J., Waldrep K., and Bernett J. The estimation of total serum lipids by a completely enzymatic summation method. Clin Chim Acta 1989: 184: 219–226.

    Article  CAS  Google Scholar 

  • Birnbaum L. The mechanism of dioxin toxicity: relationship to risk assessment. Environ Health Perspect 1994: 102: 157–167.

    Article  CAS  Google Scholar 

  • Birnbaum L. Developmental effects of dioxins and related endocrine disrupting chemicals. Toxicol Lett 1995: 82/83: 743–750.

    Article  CAS  Google Scholar 

  • Bovee T.F., Hoogenboom L.A., Hamers A.R., Traag W.A., Zuidema T., Aarts J.M., Brouwer A., and Kuiper H.A. Validation and use of the CALUX-bioassay for the determination of dioxins and PCBs in bovine milk. Food Addit Contamin 1998: 15: 863–875.

    Article  CAS  Google Scholar 

  • Brown D., Chu M., Van Overmeire I., Chu A., and Clark G. Determination of REP values for the CALUX bioassay and comparison to the WHO TEF values. Organohalogen Compds 2001a: 53: 211–214.

    CAS  Google Scholar 

  • Brown D., Goeyens L., Van Overmeire I., Chu M., Murata H., and Clark G. Quality control criteria implemented for monitoring the use of the CALUX bioassay. Organohalogen Compds 2001b: 54: 32–35.

    CAS  Google Scholar 

  • Brown D., Nakamura M., Chu M., Denison M., Murata H., and Clark G. Recovery determinations for bioassay analysis: considerations and results. Organohalogen Compds 2002a: 58: 357–360.

    CAS  Google Scholar 

  • Brown D., Van Overmeire I., Goeyens L., Chu M., Denison M., and Clark G. Elimination of interfering compounds in preparation for analysis by an Ah receptor based bioassay. Organohalogen Compds 2002b: 58: 401–404.

    CAS  Google Scholar 

  • Den Hond E., Roels H.A., and Hoppenbrouwers K., et al. Sexual maturation in relation to polychlorinated aromatic hydrocarbons: Sharpe and Skakkebaek's hypothesis revisited. Environ Health Perspect 2002: 110: 771–776.

    Article  CAS  Google Scholar 

  • Denison M. Optimization of the CALUX Bioassay for use in detecting dioxin and related chemicals in serum. Superfund Basic Research Program Brief, Number 67, 2000.

  • Denison M.S., and Heath-Pagliuso S. The Ah receptor: a regulator of the biochemical and toxicological actions of structurally diverse chemicals. Bull Environ Contamin Toxicol 1998: 61: 557–568.

    Article  CAS  Google Scholar 

  • Denison M.S., Rogers W., and Fair M., et al. Application of the CALUX bioassay system for the detection of dioxin-like chemicals (Ah receptor ligands) in whole serum samples and in extracts from commercial and consumer products. Organohalogen Compds 1996: 27: 280–284.

    CAS  Google Scholar 

  • Eskenazi B., Warner M., Bonsignore L., Olive D., Samuels S., and Vercellini P. Validation study of nonsurgical diagnosis of endometriosis. Fertil Steril 2001: 76: 929–935.

    Article  CAS  Google Scholar 

  • Falk C., Hanrahan L., and Anderson H.A., et al. Body burden levels of dioxin, furans, and PCBs among frequent consumers of Great Lakes sport fish. The Great Lakes Consortium. Environ Res 1999: 80: S19–S25.

    Article  CAS  Google Scholar 

  • Farland W.H., Schaum J., and Winters D., et al. USEPA's risk characterization of dioxin and related compounds. Organohalogen Compds 2000: 48: 248–251.

    Google Scholar 

  • Garrison P.M., Tullis K., Aarts J., Brouwer A., Giesy J.P., and Denison M.S. Species-specific recombinant cell lines as bioassay systems for the detection of 2,3,7,8-tetrachlorodibenzo-p-dioxin-like chemicals. Fundam Appl Toxicol 1996: 30: 194–203.

    Article  CAS  Google Scholar 

  • Gonzalez C.A., Kogevinas M., and Huici A., et al. Blood levels of polychlorinated dibenzodioxins, polychlorinated dibenzofurans and polychlorinated biphenyls in the general population of a Spanish Mediterranean city. Chemosphere 1998: 36 (3): 419–426.

    Article  CAS  Google Scholar 

  • Hornung R., and Reed L. Estimation of average concentration in the presence of non-detectable values. Appl Occup Environ Hyg 1990: 5: 48–51.

    Article  Google Scholar 

  • IARC. Polychlorinated Dibenzo-para-Dioxins and Polychlorinated Dibenzofurans, Vol. 69. International Agency for Research on Cancer. Lyon, France, 1997.

  • Kayama F., Hamamatsu A., Sagisaka K., Brown D., Clark G., and Suzuki T. CALUX assay is applicable to human epidemiological studies. Organohalogens Compds 2001: 54: 48–50.

    CAS  Google Scholar 

  • Kayama F., Horiguchi H., and Hoguma E., et al. Regional differences of blood dioxin and organochlorine pesticides concentrations of Japanese female farmers — application of Calux assay for epidemiological study. Organohalogen Compds 2002: 55: 275–278.

    CAS  Google Scholar 

  • Kiviranta H., Vartiainen T., and Tuomisto J. Polychlorinated dibenzo-p-dioxins, dibenzofurans, and biphenyls in fishermen in Finland. Environ Health Perspect 2002: 110: 355–361.

    Article  CAS  Google Scholar 

  • Koppen G., Covaci A., and Van Cleuvenbergen R., et al. Comparison of CALUX-TEQ values with PCB and PCDD/F measurements in human serum of the Flanders Environmental and Health Study (FLEHS). Toxicol Lett 2001: 123: 59–67.

    Article  CAS  Google Scholar 

  • Koppen G., Covaci A., and Van Cleuvenbergen R., et al. Persistent organochlorine pollutants in human serum of 50–65 years old women in the Flanders Environmental and Health Study (FLEHS). Part 1: Concentrations and regional differences. Chemosphere 2002: 48: 811–825.

    Article  CAS  Google Scholar 

  • Laier P., Cederberg T., Larsen J.C., and Vinggaard A.M. Applicability of the CALUX bioassay for screening of dioxin levels in human milk samples. Food Addit Contamin 2003: 20: 583–595.

    Article  CAS  Google Scholar 

  • Landi M.T., Consonni D., and Patterson Jr. D.G., et al. 2,3,7,8-Tetrachlorodibenzo-p-dioxin plasma levels in Seveso 20 years after the accident. Environ Health Perspect 1998: 106: 273–277.

    Article  CAS  Google Scholar 

  • Longnecker M.P., Ryan J.J., Gladen B.C., and Schecter A.J. Correlations among human plasma levels of dioxin-like compounds and polychlorinated biphenyls (PCBs) and implications for epidemiologic studies. Arch Environ Health 2000: 55: 195–200.

    Article  CAS  Google Scholar 

  • Mocarelli P., Marocchi A., and Brambilla P., et al. Human data derived from the Seveso accident-relevance for human risk assessment. Toxic Subst J 1992: 12: 51–73.

    Google Scholar 

  • Mocarelli P., Pocchiari F., and Nelson N. Preliminary report: 2,3,7,8-tetrachlorodibenzo-p-dioxin exposure to humans — Seveso, Italy. MMWR 1988, 733–736.

  • Nawrot T.S., Staessen J.A., and Den Hond E.M., et al. Host and environmental determinants of polychlorinated aromatic hydrocarbons in serum of adolescents. Environ Health Perspect 2002: 110: 583–589.

    Article  CAS  Google Scholar 

  • NIST. Certificate of Analysis Standard Reference Material 1589a. National Institute of Standards and Technology, Gaithersburg, MD, 2000.

  • Papke O. PCDD/PCDF: human background data for Germany, a 10-year experience. Environ Health Perspect 1998: 106 (Suppl 2): 723–731.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Patterson D., Hampton L., and Lapeza C., et al. High-resolution gas chromatographic/high-resolution mass spectrometric analysis of human serum on a whole-weight and lipid basis for 2,3,7,8-tetrachlorodibenzo-p-dioxin. Anal Chem 1987: 59: 2000–2005.

    Article  CAS  Google Scholar 

  • Patterson D.G., Todd G.D., and Turner W.E., et al. Levels of non-ortho-substituted (coplanar), mono- and di-ortho-substituted polychlorinated biphenyls, dibenzo-p-dioxins, and dibenzofurans in human serum and adipose tissue. Environ Health Perspect 1994: 102 (Suppl 1): 195–204.

    Article  CAS  Google Scholar 

  • Pauwels A., Cenijn P., Schepens P., and Brouwer A. Comparison of chemical-activated luciferase gene expression bioassay and gas chromatography for PCB determination in human serum and follicular fluid. Environ Health Perspect 2000: 108: 553–557.

    Article  CAS  Google Scholar 

  • Pauwels A., Schepens P.J.C., and Hooghe T.D., et al. The risk of endometriosis and exposure to dioxins and polychlorinated biphenyls: a case–control study of infertile women. Hum Reprod 2001: 16: 2050–2055.

    Article  CAS  Google Scholar 

  • Schecter A., Sheu S., Birnbaum L.S., DeVito M.J., Denison M.S., and Papke O. A comparison and discussion of two differing methods of measuring dioxin-like compounds: gas chromatography-mass spectrometry and the calux bioassay- implications for health studies. Organohalogens Compds 1999: 40: 247–253.

    CAS  Google Scholar 

  • Stata C Stata Statistical Software: Release 7.0. Stata Press, College Station, TX, 2001.

    Google Scholar 

  • Svensson B.G., Nilsson A., Hansson M., Rappe C., Akesson B., and Skerfving S. Exposure to dioxins and dibenzofurans through the consumption of fish. N Engl J Med 1991: 324: 8–12.

    Article  CAS  Google Scholar 

  • USEPA. Report on the comparison of cell-based assays and mass spectrometry methods for the analysis of PCDDs, PCDFs, and PCBs in biosolids, pp. 1–20. US Environmental Protection Agency, Office of Water, Office of Science and Technology, Engineering and Analysis Division. Washington, DC, 2002.

  • Van den Berg M., Birnbaum L., and Bosveld A.T.C. . et al. Toxic equivalency factors (TEFs) for PCBs, PCDDs, PCDFs for humans and wildlife. Environ Health Perspect 1998: 106: 775–792.

    Article  CAS  Google Scholar 

  • Van Den Heuvel R.L., Koppen G., and Staessen J.A., et al. Immunologic biomarkers in relation to exposure markers of PCBs and dioxins in Flemish adolescents (Belgium). Environ Health Perspect 2002: 110: 595–600.

    Article  CAS  Google Scholar 

  • Van Overmeire I., Clark G., and Brown D., et al. Trace contamination with dioxin-like chemicals: evaluation of bioassay-based TEQ determination for hazard assessment and regulatory responses. Environ Sci Policy 2001: 4: 345–357.

    Article  CAS  Google Scholar 

  • Van Overmeire I., Goeyens L., and Beernaert H., et al. A comparative study of GC-HRMS and CALUX TEQ determinations in food samples by the Belgiam federal ministries of public health and agriculture. Organohalogens Compds 2000: 45: 196–199.

    CAS  Google Scholar 

  • Van Wouwe N., Eppe G., and Xhrouet C., et al. Analysis of PCDD/Fs in human blood plasma using CALUX bioassay and GC-HRMS: A comparison. Organohalogen Compds 2003: 60: 211–214.

    CAS  Google Scholar 

  • Whitlock J. Mechanistic aspects of dioxin action. Chem Res Toxicol 1993: 6: 754–763.

    Article  CAS  Google Scholar 

  • Zook D, and Rappe C Environmental sources, distribution, and fate. In: Schecter A. (Ed.). Dioxins and Health. Plenum Press, New York, 1994, pp. 79–113.

    Chapter  Google Scholar 

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Acknowledgements

We gratefully acknowledge Stefania Casalini, PhD for coordinating data collection at Hospital of Desio. We would especially like to thank the women who participated in this study. This study was supported by Grant Numbers R01 ES07171 and F06 TW02075-01 from the National Institutes of Health, R82471 from the US Environmental Protection Agency, EA-M1977 from the Endometriosis Association, 2P30-ESO01896-17 from the National Institute of Environmental Health Sciences, and #2896 from Regione Lombardia and Fondazione Lombardia Ambiente, Milan, Italy.

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Correspondence to Marcella Warner.

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Warner, M., Eskenazi, B., Patterson, D. et al. Dioxin-Like TEQ of women from the Seveso, Italy area by ID-HRGC/HRMS and CALUX. J Expo Sci Environ Epidemiol 15, 310–318 (2005). https://doi.org/10.1038/sj.jea.7500407

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