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

• Published in: Analytical and bioanalytical chemistry Vol. 406; no. 8; pp. 2171 - 2178
• Main Authors: Kataoka, Hiroyuki, Inoue, Tsutomu, Ikekita, Natsuki, Saito, Keita
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.03.2014; Springer; Springer Nature B.V
• Subjects: Amines; Amines - isolation & purification; Amines - urine; Analytical Chemistry; Animals; Automation; Biochemistry; Biomarkers; Biomarkers - urine; Cancer; Carcinogens; Cattle; Characterization and Evaluation of Materials; Chemical properties; Chemistry; Chemistry and Materials Science; Chromatography; Chromatography, High Pressure Liquid - methods; Composition; Cooking; Extraction (Chemistry); Food; Food Contamination - analysis; Food Science; Heterocyclic amines; Heterocyclic compounds; Heterocyclic Compounds - analysis; Heterocyclic Compounds - isolation & purification; Humans; Identification and classification; Laboratory Medicine; Liquid chromatography; Liquids; Mass spectrometry; Meat; Meat - adverse effects; Meat - analysis; Metabolites; Methods; Microextraction Techniques; Monitoring/Environmental Analysis; Research Paper; Risk; Scientific imaging; Solid Phase Microextraction - instrumentation; Solid Phase Microextraction - methods; Spectrometry, Mass, Electrospray Ionization - methods; Tandem Mass Spectrometry - methods; Tumors; Urine; Urine; Heterocyclic amines; Burnt foods; Exposure biomarkers; Liquid chromatography–tandem mass spectrometry; In-tube solid-phase microextraction
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-013-7420-1

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. Figure 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