A gas chromatography-isotope dilution high-resolution mass spectrometry method for quantification of isomeric benzo[a]pyrene diol epoxide hemoglobin adducts in humans.
We developed a gas chromatography-isotope dilution high-resolution mass spectrometry (GC-ID-HRMS) method for quantifying isomers of benzo[a]pyrene (BaP) tetrol metabolites resulting from hydrolysis of benzo[a]pyrene-diol-epoxide hemoglobin (BaPDE-Hb) adducts. Acid hydrolysis of BPDE-Hb adducts extracted from human blood samples yielded isomers of benzo[a]pyrene-tetrahydrotetrols, (+/-)-BaP-r-7,t-8,t-9,c-10-tetrol (BPTI-1), (+/-)-BaP-r-7,t-8,t-9,t-10-tetrol (BPTI-2), (+/-)-BaP-r-7,t-8,c-9,t-10-tetrol (BPTII-1), and (+/-)-BaP-r-7,t-8,c-9,c-10-tetrol (BPTII-2). The isomeric BaP tetrols were isolated from the matrix by liquid-liquid extraction, and then further purified by solid-phase extraction. Following silylation with N-methyl-N-(trimethylsilyl)-trifluoroacetamide, the analytes were measured by GC-HRMS, using electron ionization. We have found detectable concentrations in the low fmol range for BPTII-1 and BPTI-1 in all donors tested. The mean BaP adduct levels for smoking donors (n = 9) were 0.022 fmol/mg hemoglobin for BPTII-1 and 0.070 fmol/mg hemoglobin for BPTI-1. The mean BaP adduct levels with hemoglobin for non smoking donors (n = 6) was 0.021 fmol/mg hemoglobin for BPTII-1 and 0.105 fmol/mg hemoglobin for BPTI-1.
Ragin
Angela D
AD
National Center for Environmental Health, Centers for Disease Control and Prevention, 4770 Buford Hwy, NE, Atlanta, Georgia 30341, USA. aragin@cdc.gov
Crawford
Kenroy E
KE
Etheredge
Alisha A
AA
Grainger
James
J
Patterson
Donald G
DG
D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / Q000378:metabolism*; D002138:Calibration; D002849:Chromatography, Gas; D015331:Cohort Studies; D004912:Erythrocytes / Q000737:chemistry; D006454:Hemoglobins / Q000032:analysis*; D006801:Humans; D006868:Hydrolysis; D007202:Indicators and Reagents; D013058:Mass Spectrometry; D011786:Quality Control; D011865:Radioisotope Dilution Technique; D012015:Reference Standards; D012907:Smoking / Q000097:blood
D006454:Hemoglobins; D007202:Indicators and Reagents; D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide