title: Analysis of 4-aminobiphenyl hemoglobin adducts in smokers and nonsmokers by pseudo capillary on-column gas chromatography- tandem mass spectrometry.
abstract: We describe here a hemoglobin adduct assay applied to an analysis of samples from smokers and nonsmokers. The assay includes a sensitive method for quantification of orthotoluidine 2-aminonaphthylene, and 3- and 4-aminobiphenyl hemoglobin adducts in human blood using capillary gas chromatography-tandem mass spectrometry. Basic hydrolysis and derivatization with pentafluoropropionic acid anhydride are followed by programmable temperature vaporization and pseudo on-column capillary gas chromatography with positive electron ionization tandem mass spectrometry analysis. Standard deviation of calibration curves (n = 6) shows that the limits of detection for o-toluidine, 2-aminonaphthylene, and 3- and 4-aminobiphenyl were 0.23, 0.39, 0.30, and 0.24 pg total on-column, respectively. The effective working limit of detection is estimated at approximately 5.22 pg/g Hb and 18.73 pg/g Hb for 4-aminobiphenyl and 2-aminonaphthylene, respectively. In a group that was predominately male and African-American, the level of 4-aminobiphenyl Hb adducts was significantly different between smokers and nonsmokers. Among 93 nonsmokers with serum cotinine concentrations less than 10 ng/mL, the geometric mean (95% CI) concentration of 4-aminobiphenyl was 29.9 pg/g hemoglobin (Hb; 29.4 to 30.4). Conversely, in 100 smokers the 4-aminobiphenyl adducts geometric mean concentration was significantly greater at 73.0 pg/g Hb (72.6 to 73.4). 4-Aminobiphenyl hemoglobin adduct and serum cotinine concentrations were correlated (r = 0.496; p < 0.0001; n = 193). In 15% of smokers, 3-aminobiphenyl was detected at low concentration. Adduct levels of 2-aminonaphthylene and ortho-toluidine were not significantly different between the smoker and nonsmoker participants. Our study shows that 4-aminobiphenyl Hb adducts remain the preferred biomarker for identifying people exposed to aromatic amines from tobacco smoke.
authors.affiliation: Emergency Response and Air Toxicants Branch, Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, Georgia 30341, USA. tvh2@cdc.gov
authors.forename: Tiffany H
authors.initials: TH
authors.lastname: Seyler
authors.forename: Levi-Rose
authors.initials: LR
authors.lastname: Reyes
authors.forename: John T
authors.initials: JT
authors.lastname: Bernert
mesh_terms: D000611:Aminobiphenyl Compounds / Q000097:blood* / Q000737:chemistry; D001681:Biological Assay; D002273:Carcinogens / Q000737:chemistry / Q000378:metabolism*; D002849:Chromatography, Gas / Q000379:methods*; D005260:Female; D006454:Hemoglobins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D019570:Inhalation Exposure / Q000032:analysis; D008297:Male; D012907:Smoking / Q000097:blood*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014028:Tobacco Smoke Pollution
chemical_list: D000611:Aminobiphenyl Compounds; D002273:Carcinogens; D006454:Hemoglobins; D014028:Tobacco Smoke Pollution; C006757:4-biphenylamine