Heterocyclic aromatic amines (HAA) are listed by the US Food and Drug Administration (FDA) as harmful or potentially harmful constituents (HPHCs) in tobacco smoke. In the present study, we evaluated HAA exposure levels in the general US population based on the 2013-2014 National Health and Nutrition Examination Survey (NHANES). Measurement of HAAs in urine is challenging because of low HAA concentrations and labor-intensive sample preparation. To cope with these challenges, we developed high-throughput and sensitive analytical method for urinary HAA using isotope-dilution liquid chromatography–tandem mass spectrometry and robotic sample preparation. The high-throughput robotic sample preparation system could handle over 300 samples per day, while maintaining intra-day and inter-day imprecision and bias ≤10 %. The validated method was successfully applied to measure HAAs in urine samples collected from &gt;3,000 participants of NHANES 2013-2014. Of the 10 HAA measured, urinary concentrations of both 2-amino-9H-pyrido[2,3-b]-indole (AαC) and 2-amino-3-methyl-9H-pyrido[2,3-b]-indole (MeAαC) in smokers were significantly higher than those in non-smokers. Conversely, the urinary levels of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) were similar between smokers and non-smokers, likely because PhIP exposure is primarily from consumption of well-done cooked meat. In summary, the sensitivity, precision, accuracy, and throughput of our assay are adequate for large-scale biomonitoring studies to provide population-scale data about the magnitude and prevalence of exposure to HAAs. (Acknowledge Center for Tobacco Products, FDA for providing funds for this project.)

Determination of exposure to heterocyclic aromatic amines in the general US population with sensitive tandem mass spectrometry and high-throughput robotic sample preparation

254th National Meeting (2017)

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