Skip to main content
SpringerLink
Log in

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

  • Research Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

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

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Sugimura T, Wakabayashi K, Nakagama H, Nagao M (2004) Heterocyclic amines: mutagens/carcinogens produced during cooking of meat and fish. Cancer Sci 95:290–299

    Article  CAS  Google Scholar 

  2. Knize MG, Felton JS (2005) Formation and human risk of carcinogenic heterocyclic amines formed from natural precursors in meat. Nutr Rev 63:158–165

    Article  Google Scholar 

  3. Turesky RJ (2007) Formation and biochemistry of carcinogenic heterocyclic aromatic amines in cooked meats. Toxicol Lett 168:219–227

    Article  CAS  Google Scholar 

  4. Alaejos MS, González V, Afonso AM (2008) Exposure to heterocyclic aromatic amines from the consumption of cooked red meat and its effect on human cancer risk: a review. Food Addit Contam Part A: Chem Anal Control Expo Risk Assess 25:2–24

    Article  CAS  Google Scholar 

  5. Felton JS, Knize MG, Wu RW, Colvin ME, Hatch FT, Malfatti MA (2007) Mutagenic potency of food-derived heterocyclic amines. Mutat Res 616:90–94

    Article  CAS  Google Scholar 

  6. Cheng KW, Chen F, Wang M (2006) Heterocyclic amines: chemistry and health. Mol Nutr Food Res 50:1150–1170

    Article  CAS  Google Scholar 

  7. Murkovic M (2004) Formation of heterocyclic aromatic amines in model systems. J Chromatogr B Anal Technol Biomed Life Sci 802:3–10

    Article  CAS  Google Scholar 

  8. Alaejos MS, Ayala JH, González V, Afonso AM (2008) Analytical methods applied to the determination of heterocyclic aromatic amines in foods. J Chromatogr B Anal Technol Biomed Life Sci 862:15–42

    Article  Google Scholar 

  9. Sinha R (2002) An epidemiologic approach to studying heterocyclic amines. Mutat Res 506–507:197–204

    Article  Google Scholar 

  10. Felton JS, Knize MG, Salmon CP, Malfatti MA, Kulp KS (2002) Human exposure to heterocyclic amine food mutagens/carcinogens: relevance to breast cancer. Environ Mol Mutagen 39:112–118

    Article  CAS  Google Scholar 

  11. Tasevska N, Sinha R, Kipnis V, Subar AF, Leitzmann MF, Hollenbeck AR, Caporaso NE, Schatzkin A, Cross AJ (2009) A prospective study of meat, cooking methods, meat mutagens, heme iron, and lung cancer risks. Am J Clin Nutr 89:1884–1894

    Article  CAS  Google Scholar 

  12. Turesky RJ, Le Marchand L (2011) Metabolism and biomarkers of heterocyclic aromatic amines in molecular epidemiology studies: lessons learned from aromatic amines. Chem Res Toxicol 24:1169–1214

    Article  CAS  Google Scholar 

  13. Alexander J, Reistad R, Hegstad S, Frandsen H, Ingebrigtsen K, Paulsen JE, Becher G (2002) Biomarkers of exposure to heterocyclic amines: approaches to improve the exposure assessment. Food Chem Toxicol 40:1131–1137

    Article  CAS  Google Scholar 

  14. Murray S, Lake BG, Gray S, Edwards AJ, Springall C, Bowey EA, Williamson G, Boobis AR, Gooderham NJ (2001) Effect of cruciferous vegetable consumption on heterocyclic aromatic amine metabolism in man. Carcinogenesis 22:1413–1420

    Article  CAS  Google Scholar 

  15. Kapiszewska M (2006) A vegetable to meat consumption ratio as a relevant factor determining cancer preventive diet. The Mediterranean versus other European countries. Forum Nutr 59:130–153

    Article  Google Scholar 

  16. Platt KL, Edenharder R, Aderhold S, Muckel E, Glatt H (2010) Fruits and vegetables protect against the genotoxicity of heterocyclic aromatic amines activated by human xenobiotic-metabolizing enzymes expressed in immortal mammalian cells. Mutat Res 703:90–98

    Article  CAS  Google Scholar 

  17. Abdull Razis AF, Noor NM (2013) Cruciferous vegetables: dietary phytochemicals for cancer prevention. Asian Pac J Cancer Prev 14:1565–1570

    Article  Google Scholar 

  18. Appenzeller BM, Tsatsakis AM (2012) Hair analysis for biomonitoring of environmental and occupational exposure to organic pollutants: state of the art, critical review and future needs. Toxicol Lett 210:119–140

    Article  CAS  Google Scholar 

  19. Kataoka H, Inoue T, Saito K, Kato H, Masuda K (2013) Analysis of heterocyclic amines in hair by on-line in-tube solid-phase microextraction coupled with liquid chromatography-tandem mass spectrometry. Anal Chim Acta 786:54–60

    Article  CAS  Google Scholar 

  20. Strickland PT, Oian Z, Friesen MD, Rothman N, Sinha R (2002) Metabolites of 2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine (PhIP) in human urine after consumption of charbroiled or fried beef. Mutat Res 506–507:163–173

    Article  Google Scholar 

  21. Shah FU, Barri T, Jönsson JA, Skog K (2004) Determination of heterocyclic aromatic amines in human urine by using hollow-fibre supported liquid membrane extraction and liquid chromatography-ultraviolet detection system. J Chromatogr B Anal Technol Biomed Life Sci 870:203–208

    Article  Google Scholar 

  22. Lezamiz J, Barri T, Jönsson JA, Skog K (2008) A simplified hollow-fibre supported liquid membrane extraction method for quantification of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) in urine and plasma samples. Anal Bioanal Chem 390:689–696

    Article  CAS  Google Scholar 

  23. De Andrés F, Zougagh M, Castañeda G, Sánchez-Rojas JL, Ríos A (2011) Screening of non-polar heterocyclic amines in urine by microextraction in packed sorbent-fluorimetric detection and confirmation by capillary liquid chromatography. Talanta 83:1562–1567

    Article  Google Scholar 

  24. De Andrés F, Zougagh M, Castañeda G, Ríos A (2010) Determination of heterocyclic amines in urine samples by capillary liquid chromatography with evaporated light-scattering detection. Anal Bioanal Chem 397:223–231

    Article  Google Scholar 

  25. Holland RD, Taylor J, Schoenbachler L, Jones RC, Freeman JP, Miller DW, Lake BG, Gooderham NJ, Turesky RJ (2004) Rapid biomonitoring of heterocyclic aromatic amines in human urine by tandem solvent solid phase extraction liquid chromatography electrospray ionization mass spectrometry. Chem Res Toxicol 17:1121–1136

    Article  CAS  Google Scholar 

  26. Holland RD, Gehring T, Taylor J, Lake BG, Gooderham NJ, Turesky RJ (2005) Formation of a mutagenic heterocyclic aromatic amine from creatinine in urine of meat eaters and vegetarians. Chem Res Toxicol 18:579–590

    Article  CAS  Google Scholar 

  27. Frandsen H (2008) Biomonitoring of urinary metabolites of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) following human consumption of cooked chicken. Food Chem Toxicol 46:3200–3205

    Article  CAS  Google Scholar 

  28. Busquets R, Jönsson JA, Frandsen H, Puignou L, Galceran MT, Skog K (2009) Hollow fibre-supported liquid membrane extraction and LC-MS/MS detection for the analysis of heterocyclic amines in urine samples. Mol Nutr Food Res 53:1496–1504

    Article  CAS  Google Scholar 

  29. Fede J-M, Thakur AP, Gooderham NJ, Turesky RJ (2009) Biomonitoring of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and its carcinogenic metabolites in urine. Chem Res Toxicol 22:1096–1105

    Article  CAS  Google Scholar 

  30. Gu D, Raymundo MM, Kadlubar FF, Turesky RJ (2011) Ultraperformance liquid chromatography-tandem mass spectrometry method for biomonitoring cooked meat carcinogens and their metabolites in human urine. Anal Chem 83:1093–1101

    Article  CAS  Google Scholar 

  31. Busquets R, Frandsen H, Jönsson JÅ, Puignou L, Galceran MT, Skog K (2013) Biomonitoring of dietary heterocyclic amines and metabolites in urine by liquid phase microextraction: 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), a possible biomarker of exposure to dietary PhIP. Chem Res Toxicol 26:233–240

    Article  CAS  Google Scholar 

  32. Viberg P, Wahlund KG, Skog K (2006) On-line capillary based quantitative analysis of a heterocyclic amine in human urine. J Chromatogr A 1133:347–352

    Article  CAS  Google Scholar 

  33. Sentellas S, Moyano E, Puignou L, Galceran MT (2004) Optimization of a clean-up procedure for the determination of heterocyclic aromatic amines in urine by field-amplified sample injection-capillary electrophoresis-mass spectrometry. J Chromatogr A 1032:193–201

    Article  CAS  Google Scholar 

  34. Kataoka H, Saito K (2012) Recent advances in column switching sample preparation in bioanalysis. Bioanalysis 4:809–832

    Article  CAS  Google Scholar 

  35. Kataoka H, Lord HL, Pawliszyn J (2000) Simple and rapid determination of amphetamine, methamphetamine, and their methylenedioxy derivatives in urine by automated in-tube solid-phase microextraction coupled with liquid chromatography-electrospray ionization mass spectrometry. J Anal Toxicol 24:257–265

    Article  CAS  Google Scholar 

  36. Kataoka H, Inoue R, Yagi K, Saito K (2009) Determination of nicotine, cotinine, and related alkaloids in human urine and saliva by automated in-tube solid-phase microextraction coupled with liquid chromatography-mass spectrometry. J Pharm Biomed Anal 49:108–114

    Article  CAS  Google Scholar 

  37. Saito K, Yagi K, Ishizaki A, Kataoka H (2010) Determination of anabolic steroids in human urine by automated in-tube solid-phase microextraction coupled with liquid chromatography-mass spectrometry. J Pharm Biomed Anal 52:727–733

    Article  CAS  Google Scholar 

  38. Kataoka H (2002) Automated sample preparation using in-tube solid-phase microextraction and its application—a review. Anal Bioanal Chem 373:31–45

    Article  CAS  Google Scholar 

  39. Kataoka H, Ishizaki A, Nonaka Y, Saito K (2009) Developments and applications of capillary microextraction techniques: a review. Anal Chim Acta 655:8–29

    Article  CAS  Google Scholar 

  40. Kataoka H (2010) Recent developments and applications of microextraction techniques in drug analysis. Anal Bioanal Chem 396:339–364

    Article  CAS  Google Scholar 

  41. Kataoka H, Saito K (2011) Recent advances in SPME techniques in biomedical analysis. J Pharm Biomed Anal 54:926–950

    Article  CAS  Google Scholar 

Download references

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).

Author information

Authors and Affiliations

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

    Hiroyuki Kataoka, Tsutomu Inoue, Natsuki Ikekita & Keita Saito

Authors
  1. Hiroyuki Kataoka
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tsutomu Inoue
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Natsuki Ikekita
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Keita Saito
    View author publications

    You can also search for this author in PubMed Google Scholar

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.

Electronic Supplementary Material

Below is the link to the electronic supplementary material.

ESM 1

(PDF 1114 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

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

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

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

Keywords

Search

Navigation