HPLC-accelerator MS measurement of atrazine metabolites in human urine after dermal exposure

Bruce A. Buchholz, Esther Fultz, Kurt W. Haack, John S. Vogel, S. Douglas Gilman, Shirley J. Gee, Bruce D. Hammock, Xiaoying Hui, Ronald C. Wester, Howard I. Maibach

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Abstract

Metabolites of atrazine were measured in human urine after dermal exposure using HPLC to separate and identify metabolites and accelerator mass spectrometry (AMS) to quantify them. Ring-labeled [14C]atrazine was applied for 24 h with a dermal patch to human volunteers at low (0.167 mg, 6.45 μCi) and high (1.98 mg, 24.7 μCi) doses. Urine was collected for 7 days. The urine was centrifuged to remove solids, and the supernatant was measured by liquid scintillation counting prior to injection on the HPLC to ensure that <0.17 Bq (4.5 pCi) was injected on the column. A reversed-phase gradient of 0.1% acetic acid in water and 0.1% acetic acid in acetonitrile became less polar with increasing time and separated the parent compound and major atrazine metabolites over 31 min on an octadecylsilane column. Peaks were identified by coelution with known standards. Elution fractions were collected in 1-min increments; half of each fraction was analyzed by AMS to obtain limits of quantitation of 14 amol. Mercapturate metabolites of atrazine and dealkylated atrazine dominated the early metabolic time points, accounting for ~90% of the 14C in the urine. No parent compound was detected. The excreted atrazine metabolites became more polar with increasing time, and an unidentified polar metabolite that was present in all samples became as prevalent as any of the known ring metabolites several days after the dose was delivered. Knowledge of metabolite dynamics is crucial to developing useful assays for monitoring atrazine exposure in agricultural workers.

Original languageEnglish (US)
Pages (from-to)3519-3525
Number of pages7
JournalAnalytical Chemistry
Volume71
Issue number16
DOIs
StatePublished - Aug 15 1999

ASJC Scopus subject areas

  • Analytical Chemistry

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    Buchholz, B. A., Fultz, E., Haack, K. W., Vogel, J. S., Gilman, S. D., Gee, S. J., Hammock, B. D., Hui, X., Wester, R. C., & Maibach, H. I. (1999). HPLC-accelerator MS measurement of atrazine metabolites in human urine after dermal exposure. Analytical Chemistry, 71(16), 3519-3525. https://doi.org/10.1021/ac990152g