High-throughput and sensitive analysis of urinary heterocyclic aromatic amines using isotope-dilution liquid chromatography–tandem mass spectrometry and robotic sample preparation system

Abstract

Heterocyclic aromatic amines (HCAA) are listed by the US Food and Drug Administration (FDA) as harmful or potentially harmful constituents of tobacco smoke. However, quantifying HCAA exposure is challenging. In this study, we developed a sensitive, precise and accurate isotope dilution, liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to quantify urinary HCAAs in smokers and nonsmokers. The high throughput robotic sample preparation system could handle a throughput of over 300 samples per day, while maintaining intra-day and inter-day imprecision and bias ≤10%. The limits of detection of carcinogenic HCAAs ranged from 0.31-0.83 pg/mL. The validated method was applied to measure HCAAs in urine collected from smokers and non-smokers. This sensitive and efficient analytical method is ideal to support large-scale biomonitoring studies of HCAA exposure.

Metabolic Evidence Rather Than Amounts of Red or Processed Meat as a Risk on Korean Colorectal Cancer

“…The cases were newly diagnosed with CRC; thus, most were at early stages of the disease, and most locations of CRC were distal ( Table S1 ). We also studied some confounders for the exposure biomarkers and metabolites, e.g., tobacco smoking and fish intake, which may induce HCAs [ 27 ] and ω-3 fatty acids [ 28 ], respectively. These two factors were not different between the two groups ( Table 1 ).…”

Evaluation of Tobacco Smoke and Diet as Sources of Exposure to Two Heterocyclic Aromatic Amines for the U.S. Population: NHANES 2013–2014

“…Spot urine samples were stored at À70 C before assay. The total urinary concentrations of AaC and MeAaC (free and conjugated forms) were analyzed using isotope-dilution LC/MS/MS and robotic sample preparation (46). Briefly, an aliquot of 0.5 mL of urine samples was spiked with internal standards that are stable isotope analogs of targeted analytes.…”

“…After hydrolysis, the total urinary AaC and MeAaC were extracted from urine matrix by using diatomaceous earth and mixed mode of cation exchange solid phase extraction plates, respectively. The throughput and precision of sample preparation was improved through automated solid phase extraction using an integrated robotic system (46). Analytes were chromatographically resolved from urinary interferences using a reversed phase column (Agilent Zorbax Eclipse Plus C18, 2.1 Â 100 mm 3.5 mm).…”

Determination of 14 heterocyclic aromatic amines in meat products using solid‐phase extraction and supercritical fluid chromatography coupled to triple quadrupole mass spectrometry

“…Although lacking evidence to support the hypothesis that excessive uptake of HAs could directly cause cancer, the International Agency for Research on Cancer has already decided a limitation of HAs . HAs have been discovered in different matrices . Most of the researchers have isolated and identified them in food samples and related substances.…”

“…For instance, barbecue, roast fish, cooking residues (such as pan residues and cooking smoke), alcoholic beverages, and many other compounds have been reported to contain trace of HAs . Besides, in biological samples (such as serum, hair, bile, human milk, urine, and so on), HAs have been detected . Even in many environmental samples (such as rain, water, air, cigarette smoke, and so on), HAs residues have also been reported .…”