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Within-person variability in urinary phthalate metabolite concentrations: measurements from specimens after long-term frozen storage

Journal of Exposure Science & Environmental Epidemiology volume 20, pages 169–175 (2010)Cite this article

Abstract

Laboratory studies show that exposure to phthalates during development can cause adverse effects, especially for males. Studies in humans would be facilitated by collection of urine during pregnancy, long-term storage, and measurement of phthalate metabolites at the time that offspring health is assessed. Our aims were to measure urinary phthalate metabolites after long-term freezer storage, to use those measurements to evaluate within-woman variability over 2- and 4-week intervals, and to determine whether the phases of the menstrual cycle affect metabolite levels. Samples were selected from daily first-morning urine specimens collected by 60 women and stored frozen since 1983–1985. Three specimens per woman were selected at approximately 2-week intervals to include both follicular and luteal phase samples. Seven metabolites of five phthalates were measured by mass spectrometry. Statistical analyses were conducted with correlation, mixed model regression, and the Wilcoxon signed rank test. Creatinine-corrected urinary phthalate metabolite concentrations measured in samples after long-term storage tended to have a similar right-skewed distribution, though with somewhat higher concentrations than those reported for recently collected US samples. The concentrations of three metabolites of di(2-ethylhexyl)phthalate in the same specimen were very highly correlated (Pearson r=0.85–0.97). Reproducibility over a 4-week interval was moderate for the metabolites of diethyl phthalate and benzylbutyl phthalate (intraclass correlation coefficients, ICCs, 0.48 and 0.53, respectively), whereas five other metabolites had lower ICCs (0.21–0.37). Menstrual phase was not related to metabolite concentrations. Although the same samples have not been measured both before and after long-term storage, results suggest that the measurement of phthalate metabolites after long-term sample storage yield generally similar distributions and temporal reliability as those reported for recently collected specimens. These findings support the use of stored urine specimens collected during the relevant stage of human pregnancy to investigate the influence of phthalate exposures on later outcomes.

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References

  • Adibi J.J., Whyatt R.M., Williams P.L., Calafat A.M., Camann D., Herrick R., Nelson H., Bhat H.K., Perera F.P., Silva M.J., and Hauser R. Characterization of phthalate exposure among pregnant women assessed by repeat air and urine samples. Environ Health Perspect 2008: 116: 467–473.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Anderson W.A., Castle L., Scotter M.M., Massey R.C., and Springall C. A biomarker approach to measuring human dietary exposure to certain phthalate diesters. Food Addit Contam 2001: 18: 1068–1074.

    Article  CAS  PubMed  Google Scholar 

  • Baird D.D., Weinberg C.R., Wilcox A.J., McConnaughey D.R., and Musey P.I. Using the ratio of urinary estrogen and progesterone metabolites to estimate day of ovulation. Statist Med 1991: 10: 255–266.

    Article  CAS  Google Scholar 

  • Barr D.B., Wang R.Y., and Needham L.L. Biologic monitoring of exposure to environmental chemicals throughout the life stages: requirements and issues for consideration for the National Children's Study. Environ Health Perspect 2005: 113: 1083–1091.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bizzari S., Oppenberg B., and Iskikawa Y. Plasticizers. Chemical Economics Handbook. SRI International, Palo Alto, CA, USA, 2000.

    Google Scholar 

  • CDC. National Report on Human Exposure to Environmental Chemicals, Third Report. Centers for Disease Control and Prevention, Atlanta, GA, 2005.

  • Foster P.M.D. Disruption of reproductive development in male rate offspring following in utero exposure to phthalate esters. Int J Androl 2006: 29: 140–147.

    Article  CAS  PubMed  Google Scholar 

  • Frederiksen H., Skakkebaik N.E., and Andersson A.-M. Metabolism of phthalates in humans. Mol Nutr Food Res 2007: 57: 899–911.

    Article  Google Scholar 

  • Fromme H., Bolte G., Koch H.M., Angerer J., Boehmer S., Drexter H., Mayer R., and Liebl B. Occurrence and daily variation of phthalate metabolites in the urine of an adult population. Int J Hyg Environ Health 2007: 210: 21–33.

    Article  CAS  PubMed  Google Scholar 

  • Gray Jr L.E., Ostby J., Furr J., Price M., Veeramachaneni D.N.R., and Parks L. Perinatal exposure to the phthalates DEHP BBP, and DINP, but not DEP DMP, or DOTP, alters sexual differentiation of the male rat. Toxicol Sci 2000: 58: 350–365.

    Article  CAS  PubMed  Google Scholar 

  • Hauser R., and Calafat A.M. Phthalates and human health. Occup Environ Med 2005: 62: 806–818.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hauser R., Meeker J.D., Park S., Silva M.J., and Calafat A.M. Temporal variability of urinary phthalate metabolite levels in men of reproductive age. Environ Health Perspect 2004: 112: 1734–1740.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Helm D. Correlation between production amounts of DEHP and daily intake. Sci Total Environ 2007: 388: 389–391.

    Article  CAS  PubMed  Google Scholar 

  • Heudorf U., Mersch-Sundermann V., and Angerer J. Phthalates: toxicology and exposure. Int J Hyg Environ Health 2007: 210: 623–634.

    Article  CAS  PubMed  Google Scholar 

  • Hoppin J.A., Brock J.W., Davis B.J., and Baird D.D. Reproducibility of urinary phthalate metabolites in first morning urine samples. Environ Health Perspect 2002: 110: 515–518.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hoppin J.A., Ulmer R., Calafat A.M., Barr D.B., Baker S.V., Meltzer H.M., and Ronningen K.S. Impact of urine preservation methods and duration of storage on measured levels of environmental contaminants. J Expo Sci Environ Epidemiol 2005: 16: 39–48.

    Article  Google Scholar 

  • Hormung R.W., and Reed L.D. Estimation of average concentration in the presence of nondetectable values. Appl Occup Environ Hyg 1990: 5: 46–51.

    Article  Google Scholar 

  • Howdeshell K.L., Wilson V.S., Furr J., Lambright C.R., Rider C.V., Blystone C.R., Hotchkiss A.K., and Gray Jr L.E. A mixture of five phthalate esters inhibits fetal testicular testosterone production in the Sprague Dawley rat in a cumulative, dose additive manner. Toxicol Sci 2008: 105: 153–165.

    Article  CAS  PubMed  Google Scholar 

  • Huang P.-C., Kuo P.-L., Guo Y.-L., Liao P.-C., and Lee C.-C. Associations between urinary phthalate monoesters and thyroid hormones in pregnant women. Human Reprod 2007: 22: 2715–2722.

    Article  CAS  Google Scholar 

  • Koch H.M., Bolt H.M., Preuss R., and Angerer J. New metabolites of di(2-ethylhexyl)phthalate (DEHP) in human urine and serum after single oral doses of deuterium-labelled DEHP. Arch Toxicol 2005: 79: 367–376.

    Article  CAS  PubMed  Google Scholar 

  • Kolarik B., Naydenov K., Larsson M., Bornehag C.-G., and Sundell J. The association between phthalate in dust and allergic diseases among Bulgarian children. Environ Health Perspect 2008: 116: 98–103.

    Article  CAS  PubMed  Google Scholar 

  • Koo H.J., and Lee B.M. Estimated exposure to phthalates in cosmetics and risk assessment. J Toxicol Environ Health, Part A 2004: 67 (23–24): 1901–1914.

    Article  CAS  Google Scholar 

  • Main K.M. Phthalate monoesters and infant reproductive health. Gesundheitswesen 2008: 70 (Suppl 1): S46–S48.

    Article  PubMed  Google Scholar 

  • Marsee K., Woodruff T.J., Axelrad D.A., Calafat A.M., and Swan S.H. Estimated daily phthalate exposures in a population of mothers of male infants exhibiting reduced anogenital distance. Environ Health Perspect 2006: 114: 805–809.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Matsumoto M., Hirata-Koizumi H., and Ema M. Potential adverse effects of phthalic acid esters on human health: a review of recent studies on reproduction. Reg Toxicol Pharmacol 2008: 50: 37–49.

    Article  CAS  Google Scholar 

  • NTP-CERHR Expert Panel Report. Butyl Benzyl phthalate. Center for the Evaluation of Risks to Human Reproduction. National Toxicology Program. NTP-CERHR-BBP-00. 2000.

  • Schettler T. Human exposure to phthalates via consumer products. Int J Androl 2006: 29: 134–139.

    Article  CAS  PubMed  Google Scholar 

  • Shrout P., and Fleiss J. Intraclass correlations: uses in assessing rater reliability. Psychol Bull 1979: 86: 420–428.

    Article  CAS  PubMed  Google Scholar 

  • Silva M.J., Barr D.B., Reidy J.A., Malek N.A., Hodge C.C., Caudill S.P., Brock J.W., Needham L.L., and Calafat A.M. Urinary levels of seven phthalate metabolites in the U.S. population from the National Health and Nutrition Examination Survey (NHANES) 1999–2000. Environ Health Perspect 2004: 112: 331–338.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Taussky H.H. A microcolorimetric determination of creatine in urine by the Jaffe reaction. J Biol Chem 1954: 208: 853–861.

    CAS  PubMed  Google Scholar 

  • Teitelbaum S.L., Britton J.A., Calafat A.M., Ye X., Silva M.J., Reidy J.A., Galvez M.P., Brenner B.L., and Wolf M.S. Temporal variability in urinary concentrations of phthalate metabolites, phytoestrogens and phenols among minority children in the United States. Environ Res 2008: 107: 257–269.

    Article  Google Scholar 

  • Wilcox A.J., Weinberg C.R., O′Connor J.F., Baird D.D., Schlatterer J.P., Canfield R.E., Armstrong E.G., and Nisula B.C. Incidence of early loss of pregnancy. New Engl J Med 1988: 319: 189–194.

    Article  CAS  PubMed  Google Scholar 

  • Wittassek M., Wiesmuller G.A., Koch H.M., Eckard R., Dobler L., Muller J., Angerer J., and Schluter C. Internal phthalate exposure over the last two decades—a retrospective human biomonitoring study. Int J Hyg Environ Health 2007: 210: 319–333.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

The field manager of the North Carolina Early Pregnancy Study was Joy Pierce, and D. Robert McConnaughey manages the study data files. This research was funded by the intramural program at the National Institute of Environmental Health Sciences, National Institutes of Health, HHS. We thank AXYS Analytical Services Ltd from Sidney, BC Canada, for their assistance with this project including their detailed description of the analytic methods. An earlier version of the manuscript was reviewed by Drs. Freya Kamel and Yang Cao.

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Authors and Affiliations

  1. Epidemiology Branch, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina, USA

    Donna Day Baird, Jane A Hoppin, Matthew P Longnecker & Allen J Wilcox

  2. Social & Scientific Systems, Inc., Durham, North Carolina, USA

    Tina M Saldana

  3. Faculty of Health Sciences, Simon Fraser University, BC, Canada

    Pablo A Nepomnaschy

  4. Biostatistics Branch, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina, USA

    Clarice R Weinberg

Authors
  1. Donna Day Baird
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  2. Tina M Saldana
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  3. Pablo A Nepomnaschy
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  4. Jane A Hoppin
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  5. Matthew P Longnecker
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  6. Clarice R Weinberg
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  7. Allen J Wilcox
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Corresponding author

Correspondence to Donna Day Baird.

Appendix: Urinary Phthalate Measurement Information

Appendix: Urinary Phthalate Measurement Information

Table 1

Table a1 Analytes, ions, and quantification references.
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Table 2

Table a2 LC-MS/MS operating conditions for analysis.
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Baird, D., Saldana, T., Nepomnaschy, P. et al. Within-person variability in urinary phthalate metabolite concentrations: measurements from specimens after long-term frozen storage. J Expo Sci Environ Epidemiol 20, 169–175 (2010). https://doi.org/10.1038/jes.2009.17

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