Carcinogenesis, Vol. 20, No. 6, 1055-1061,
June 1999
© 1999 Oxford University Press
Carcinogenesis |
Quantitative analysis of 4-aminobiphenyl-C8-deoxyguanosyl DNA adducts produced in vitro and in vivo using HPLC–ES-MS
National Center for Toxicological Research, Jefferson, AR 72079, USA and
1 Instituto Superior Tecnico, P-1049-001 Lisboa, Portugal
Electrospray mass spectrometry (ES-MS) is a powerful tool for analysis of carcinogen-adducted DNA. In this study, we developed a quantitative isotope dilution method for analysis of N-(deoxyguanosine-8-yl)-4-aminobiphenyl (dG-C8-4-ABP), the principal nucleoside adduct derived from enzymatic hydrolysis of 4-aminobiphenyl (4-ABP)-modified DNA. The method used column switching valves to perform on-line sample concentration and cleanup, which permitted direct analysis of enzymatic DNA hydrolysates using narrow-bore liquid chromatography (LC). ES-MS detection was performed using a single quadrupole instrument by monitoring M+H+ and two fragment ions characteristic for dG-C8-4-ABP, along with M+H+ and a fragment ion for the deuterated internal standard. The detection limit for dG-C8-4-ABP in DNA hydrolysates was ~10 pg on-column, equivalent to 0.7 dG-C8-4-ABP adducts in 107 normal nucleotides for a sample containing 100 µg DNA. The method was applied to the analysis of calf thymus DNA modified in vitro through reaction with N-hydroxy-4-ABP and of hepatic DNA isolated from mice treated in vivo with two dose levels of 4-ABP.
Abbreviations: 4-ABP, 4-aminobiphenyl; ACN, acetonitrile; DELFIA, dissociation-enhanced lanthanide fluoroimmunoassay; dG, 2'-deoxyguanosine; dG-C8-4-ABP, N-(deoxyguanosine-8-yl)-4-aminobiphenyl; ELISA, enzyme-linked immunosorbent assay; ES, electrospray; ethenoGua, N-2,3-ethenoguanine; Gua, guanine; IAC, immunoaffinity chromatography; M1G, 3-(2-deoxy-ß-D-erythro-pentofuranosyl)-pyrimido[1,2-
]purin-10(3H)-one; MS, mass spectrometry; NICI, negative ion chemical ionization; RIA, radioimmunoassay; SIR, selected ion recording.
2 To whom correspondence should be addressed Email: ddoerge{at}nctr.fda.gov
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  B. Zayas, S. W. Stillwell, J. S. Wishnok, L. J. Trudel, P. Skipper, M. C. Yu, S. R. Tannenbaum, and G. N. Wogan Detection and quantification of 4-ABP adducts in DNA from bladder cancer patients Carcinogenesis, February 1, 2007; 28(2): 342 - 349. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Singh and P. B. Farmer Liquid chromatography-electrospray ionization-mass spectrometry: the future of DNA adduct detection Carcinogenesis, February 1, 2006; 27(2): 178 - 196. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. C. Chang, M. I. Churchwell, K. B. Delclos, R. R. Newbold, and D. R. Doerge Mass Spectrometric Determination of Genistein Tissue Distribution in Diet-Exposed Sprague-Dawley Rats J. Nutr., August 1, 2000; 130(8): 1963 - 1970. [Abstract] [Full Text]
|
|