title: Detection of 7-(2'-carboxyethyl)guanine but not 7-carboxymethylguanine in human liver DNA.
abstract: 7-Carboxymethylguanine (7-CMGua) and 7-(2'-carboxyethyl)guanine (7-CEGua) are DNA adducts that potentially could be formed upon the metabolism of the carcinogenic nitrosamines N-nitrososarcosine (NSAR) and 3-(methylnitrosamino)propionic acid (MNPA), respectively, or from other sources such as nitrosation of glycine (7-CMGua) or reaction of DNA with acrylic acid (7-CEGua). Since both NSAR and MNPA have been detected in human urine and there are plausible sources of exposure to other precursors to these adducts, we analyzed human liver DNA for 7-CMGua and 7-CEGua, using liquid chromatography-electrospray ionization-tandem mass spectrometry-selected reaction monitoring (LC-ESI-MS/MS-SRM). Human hepatic DNA was mixed with [15N5]7-CMGua and [15N5]7-CEGua as internal standards and enzymatically hydrolyzed. The hydrolysate was partially purified by solid-phase extraction, and the resulting fraction was treated with acetyl chloride in methanol to convert 7-CMGua and 7-CEGua to their methyl esters. After a second solid-phase extraction, LC-ESI-MS/MS-SRM analysis was carried out using the transitions m/z 224 [M + H](+) --> m/z 164 [(M + H)-HCOOCH3]+ and m/z 238 [M + H]+ --> m/z 152 [BH]+ for the methyl esters of 7-CMGua and 7-CEGua, respectively. The method was sensitive, accurate, precise, and apparently free from artifact formation. 7-CEGua, as its methyl ester, was detected in all 24 human liver samples analyzed, mean +/- SD, 373 +/- 320 fmol/mumol Gua (74.6 adducts per 10(9) nucleotides), range 17-1189 fmol/mumol Gua, but the methyl ester of 7-CMGua was not detected in any sample. These results demonstrate the ubiquitous presence of 7-CEGua in human liver DNA. Acrylic acid may be a likely endogenous precursor to 7-CEGua.
authors.affiliation: Masonic Cancer Center, University of Minnesota, MMC 806, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA.
authors.forename: Guang
authors.initials: G
authors.lastname: Cheng
authors.forename: Mingyao
authors.initials: M
authors.lastname: Wang
authors.forename: Peter W
authors.initials: PW
authors.lastname: Villalta
authors.forename: Stephen S
authors.initials: SS
authors.lastname: Hecht
mesh_terms: D002853:Chromatography, Liquid; D004247:DNA / Q000737:chemistry*; D018736:DNA Adducts / Q000032:analysis* / Q000737:chemistry; D006147:Guanine / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry; D006801:Humans; D008099:Liver / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D018736:DNA Adducts; D006147:Guanine; C034100:7-carboxymethylguanine; D004247:DNA