Development of a 32 P-postlabeling method for the detection of 1,N 2 - propanodeoxyguanosine adducts of 2-hexenal in vivo

D. Schuler, Budiawan, E. Eder

Research output: Contribution to journalArticlepeer-review

19 Citations (Scopus)

Abstract

2-Hexenal is an α,β-unsaturated carbonyl compound which forms cyclic 1,N 2 -propano adducts in vitro. The adduct formation in vivo was not reported by others to date. Because this type of adduct is considered promutagenic (2- hexenal is actually mutagenic and genotoxic) and humans are permanently exposed to this compound via vegetarian food, 2-hexenal may play a role in carcinogenicity. To improve the cancer risk assessment, we developed a new 32 P-postlabeling technique for this compound and optimized the different steps of the postlabeling procedure. The results of the postlabeling methods are shown. A labeling efficiency of 35%, a recovery of 10% for the synthesized standards, and a detection limit of three 2-hexenal adducts per 10 8 nucleotides was achieved. After gavage of 500 mg/kg of body weight to male Fischer 344 rats, the respective DNA adducts were detected in rat liver DNA. With this study, we demonstrate in vivo adduct formation of 2-hexenal for the first time. Highest adduct levels were found 2 days after gavage, and after 4 days, the level was even higher than after 1 day. No adducts were detected 8 h after gavage. The respective adducts could not be found as a background in tissues of untreated rats or in calf thymus DNA at the limit of detection.

Original languageEnglish
Pages (from-to)335-340
Number of pages6
JournalChemical Research in Toxicology
Volume12
Issue number4
DOIs
Publication statusPublished - 1 Apr 1999

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