Development of an on-line liquid chromatography-electrospray tandem mass spectrometry assay to quantitatively determine 1,N(2)-etheno-2'-deoxyguanosine in DNA.
A method involving on-line reversed-phase high-performance liquid chromatography with electrospray tandem mass spectrometry detection has been developed for the analysis of 1,N(2)-etheno-2'-deoxyguanosine in DNA. This methodology permits direct quantification of 20 fmol (7.4 adducts/10(8) dGuo) of the etheno adduct from approximately 350 microg of crude DNA hydrolysate. Using the newly developed technique, basal levels of 1,N(2)-etheno-2'-deoxyguanosine were determined in commercial calf thymus DNA (1.70 +/- 0.09 adducts/10(7) dGuo), in cultured mammalian cells (CV1-P) DNA (4.5 +/- 0.4 adducts/10(7) dGuo), and in untreated female rat liver DNA (5.22 +/- 1.37 adduct/10(7) dGuo). The mutagenicity of 1,N(2)-etheno-2'-deoxyguanosine had already been demonstrated by in vitro and in vivo systems. The method described here provides the first evidence of the occurrence of 1,N(2)-etheno-2'-deoxyguanosine as a basal endogenous lesion and may be usefully employed to assess the biological consequences of etheno DNA damage under normal and pathological conditions.
Loureiro
Ana Paula M
AP
Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, Avenida Prof. Lineu Prestes, 748, CEP-05508-900, São Paulo, São Paulo, Brazil.
Marques
Sabrina A
SA
Garcia
Camila C M
CC
Di Mascio
Paolo
P
Medeiros
Marisa H G
MH
D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D018736:DNA Adducts / Q000032:analysis*; D004249:DNA Damage; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000032:analysis*; D005260:Female; D051381:Rats; D017208:Rats, Wistar; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
D018736:DNA Adducts; C054579:1,N(2)-ethenodeoxyguanosine; D003849:Deoxyguanosine