title: Ultra-performance liquid chromatography-tandem mass spectrometry for rapid and highly sensitive analysis of stereoisomers of benzo[a]pyrene diol epoxide-DNA adducts.
abstract: An ultra-performance liquid chromatography tandem mass spectrometry with multiple reaction monitoring method (UPLC-MRM/MS) is developed for fast and sensitive analysis of four genotoxic stereoisomers of anti-benzo[a]pyrene diol epoxide (BPDE)-N(2)dG adducts (trans-(+), trans-(-), cis-(+) and cis-(-)), which result from environmental exposure to ubiquitous pollutant benzo[a]pyrene (B[a]P). The developed method displays a low limit of detection of <0.7 fmol (S/N=3) for the four stereoisomers of anti-BPDE-N(2)dG, a dynamic range of 2 orders of magnitude (2.3-630 fmol, R(2)> or =0.997), and one separation of 2-4 min. The developed method enables us to use the stereoisomers of anti-BPDE-N(2)dG as a biomarker and to study the stereoselectivity of metabolic activation of B[a]P in human lung A549 cells. The UPLC-MRM/MS analysis of cellular DNA exposed to B[a]P show that activation of B[a]P in A549 cells predominantly induces trans-(+)-anti-BPDE-N(2)dG with cis-(+)-anti-BPDE-N(2)dG and one syn-BPDE-N(2)dG as two minorities, while trans-(-)-anti-BPDE-N(2)dG and cis-(-)-anti-BPDE-N(2)dG are absent. The observed preferential formation of trans-(+)-anti-BPDE-N(2)dG in B[a]P treated A549 cells may result from combined stereoselectivity of the metabolic activation of B[a]P and the reaction of anti-BPDE with dsDNA. The results also suggest that a number of key optical intermediates are formed during activation of B[a]P in A549 cells, including trans-(+)-B[a]P-7,8-dihydrodiol and trans-(-)-B[a]P-7,8-dihydrodiol and their corresponding downstream metabolites (+)-anti-BPDE and (+)-syn-BPDE.
authors.affiliation: State Key laboratory of Environmental Chemistry and Eco-toxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Feng
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Wang
authors.forename: Hancheng
authors.initials: H
authors.lastname: Yuan
authors.forename: Hailin
authors.initials: H
authors.lastname: Wang
mesh_terms: D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / Q000031:analogs & derivatives / Q000737:chemistry* / Q000494:pharmacology; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D018736:DNA Adducts / Q000737:chemistry*; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000494:pharmacology; D006801:Humans; D009153:Mutagens / Q000494:pharmacology; D012680:Sensitivity and Specificity; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D018736:DNA Adducts; D009153:Mutagens; C043931:benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide-DNA; D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; C040060:7,8-dihydroxy-9,10-epoxide-7,8,9,10-tetrahydrobenzo(a)pyrene-10-deoxyguanosine; D003849:Deoxyguanosine