Screening for DNA adducts in ovarian follicles exposed to benzo[a]pyrene and cigarette smoke condensate using liquid chromatography-tandem mass spectrometry.
A rapid mass spectrometric method was applied to non-targeted screening of DNA adducts in follicular cells (granulosa cells and theca cells) from isolated ovarian follicles that were exposed in-vitro to benzo[a]pyrene (B[a]P) and cigarette smoke condensate (CSC) for 13days of culture. The method employed a constant neutral loss (CNL) scan to identify chromatographic peaks associated to a neutral loss of deoxyribose moiety of DNA nucleosides. These peaks were subsequently analyzed by a product ion scan in tandem mass spectrometry to elucidate structures of DNA adducts. The identification was further confirmed through synthesis of proposed DNA adducts where possible. Three DNA adducts, benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide-dG (BPDE-dG), phenanthrene 1,2-quinone-dG (PheQ-dG) and B[a]P-7,8-quinone-dG (BPQ-dG) were identified in the follicular cells from isolated ovarian follicles exposed to B[a]P. Along with these three, an additional DNA adduct, 4-aminobiphenyl-dG, was identified in the follicular cells from isolated ovarian follicles exposed to CSC. The amounts of the identified DNA adducts in follicular cells increased in a dose-dependent manner for both B[a]P (0, 1.5, 5, 15 and 45ng/mL) and CSC (0, 30, 60, 90 and 130μg/mL). The results revealed that B[a]P-related DNA adducts were the major adducts in the ovarian follicular cells exposed to CSC. The results also revealed that two oxidative biomarkers, 8-hydroxy-2-deoxy guanosine (8-OH-dG) and 8-isoprostane (8-IsoP), in both B[a]P-exposed and CSC-exposed ovarian follicles had strong correlations with the three DNA adducts, BPDE-dG, BPQ-dG and PheQ-dG. A pathway to describe formation of DNA adducts was proposed based on the DNA adducts observed.
Exposure and Biomonitoring Division, Environmental Health Science and Research Bureau, Healthy Environments and Consumer Safety Branch, Health Canada, 50 Colombine Driveway, AL: 0800C, Ottawa, Ontario K1A 0K9, Canada.
Chunhe
C
Yao
Department of Obstetrics & Gynecology, Michael G. DeGroote School of Medicine, McMaster University, Hamilton, Ontario, Canada.
Warren G
WG
Foster
Department of Obstetrics & Gynecology, Michael G. DeGroote School of Medicine, McMaster University, Hamilton, Ontario, Canada.
Jean C
JC
Sadeu
Exposure and Biomonitoring Division, Environmental Health Science and Research Bureau, Healthy Environments and Consumer Safety Branch, Health Canada, 50 Colombine Driveway, AL: 0800C, Ottawa, Ontario K1A 0K9, Canada.
Shabana
S
Siddique
Exposure and Biomonitoring Division, Environmental Health Science and Research Bureau, Healthy Environments and Consumer Safety Branch, Health Canada, 50 Colombine Driveway, AL: 0800C, Ottawa, Ontario K1A 0K9, Canada.
Jiping
J
Zhu
Exposure and Biomonitoring Division, Environmental Health Science and Research Bureau, Healthy Environments and Consumer Safety Branch, Health Canada, 50 Colombine Driveway, AL: 0800C, Ottawa, Ontario K1A 0K9, Canada. Electronic address: yong-lai.feng@canada.ca.
Yong-Lai
YL
Feng
D001564:Benzo(a)pyrene / Q000633:toxicity*; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D000073865:Cigarette Smoking; D018736:DNA Adducts / Q000032:analysis*; D005260:Female; D006801:Humans; D006080:Ovarian Follicle / Q000166:cytology / Q000187:drug effects*; D012906:Smoke / Q000009:adverse effects*; D053719:Tandem Mass Spectrometry
D018736:DNA Adducts; D012906:Smoke; D001564:Benzo(a)pyrene
Benzo[a]pyrene; Cigarette smoke condensate; DNA adducts; Non-targeted screening analysis; Tandem mass spectrometry