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Development of exposure assessment method based on the analysis of urinary heterocyclic amines as biomarkers by on-line in-tube solid-phase microextraction coupled with liquid chromatography–tandem mass spectrometry

Analytical and Bioanalytical Chemistry volume 406, pages 2171–2178 (2014)Cite this article

Abstract

Heterocyclic amines (HCAs) formed in cooked meats and fish are mutagens and carcinogens in rodents and nonprimates. Exposure to HCAs may also be a risk factor for human tumors, but the association between dietary intake and human cancer risk has not been determined. To assess recent exposure to HCAs, we developed a simple and sensitive method for measuring HCAs in urine by automated on-line in-tube solid-phase microextraction (SPME) using a Supel-Q PLOT capillary column as an extraction device, in combination with liquid chromatography–tandem mass spectrometry (LC–MS/MS). Thirteen HCAs were separated within 15 min using a ZORBAX Eclipse XDB-C8 column and detected selectively by multiple reaction monitoring using MS/MS. This method can be applied easily to the analysis of small amounts of urine samples without any other pretreatment except for alkaline hydrolysis of bound forms of HCAs. The quantification limits of HCAs in 0.2 mL of urine samples were about 1.7–4.1 pg/mL (S/N = 10). Using this method, we evaluated the exposure to HCAs in persons who consumed well-done pan-fried beef and the suitability of using urinary HCAs as exposure biomarkers. We also analyzed the ability of vegetable consumption to prevent carcinogenic risks from exposure to HCAs by measuring free and bound forms of HCAs in urine.

Mutagenic and carcinogenic heterocyclic amines are ingested from cooked foods and cigarette smoke, formed metabolites and adducts in target tissue, and excreted in urine and feces

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Acknowledgments

This work was supported by the National Cancer Center Research and Development Fund and the Science Research Promotion Fund, a Grant-in-Aid for Basic Scientific Research (C no. 22590048).

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Affiliations

  1. School of Pharmacy, Shujitsu University, Nishigawara, Okayama, 703-8516, Japan

    Hiroyuki Kataoka, Tsutomu Inoue, Natsuki Ikekita & Keita Saito

Authors
  1. Hiroyuki Kataoka
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  2. Tsutomu Inoue
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  3. Natsuki Ikekita
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  4. Keita Saito
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Corresponding author

Correspondence to Hiroyuki Kataoka.

Additional information

Published in the topical collection Microextraction Techniques with guest editors Miguel Valcárcel Cases, Soledad Cárdenas Aranzana and Rafael Lucena Rodríguez.

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Kataoka, H., Inoue, T., Ikekita, N. et al. Development of exposure assessment method based on the analysis of urinary heterocyclic amines as biomarkers by on-line in-tube solid-phase microextraction coupled with liquid chromatography–tandem mass spectrometry. Anal Bioanal Chem 406, 2171–2178 (2014). https://doi.org/10.1007/s00216-013-7420-1

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  • DOI: https://doi.org/10.1007/s00216-013-7420-1

Keywords

  • Heterocyclic amines
  • Urine
  • Exposure biomarkers
  • Burnt foods
  • In-tube solid-phase microextraction
  • Liquid chromatography–tandem mass spectrometry