Quantification of 1,N-6-etheno-2 '-deoxyadenosine in human urine by column-switching LC/APCI-MS/MS

Peter René Hillestrøm, A. M. Hoberg, A. Weimann, H. E. Poulsen

Research output: Contribution to journalJournal articleResearchpeer-review


1,N-6-Etheno-2'-deoxyadenosine (epsilondA) is one of several promutagenic DNA modifications arising from cellular oxidative metabolism. It is believed that these background DNA lesions may contribute to various diseases, such as cancer. Therefore, human biomonitoring of epsilondA in urine could be a potential marker for oxidative stress-related DNA damage. Existing methods for quantifying urinary edA use P-32 postlabeling. We have developed a nonradioactive, fast, and easier method based on column-switching liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry (LC/APCI-MS/MS) in the positive mode. Differences in column temperatures were used to influence analyte retention and sample focusing. With multiple reaction monitoring (MRM) mode the afforded limit of detection was about 0.7 pM when starting with 3 ml of urine. The urinary excretion rates of edA from 28 nonsmoking and 5 smoking men were 10.0-99.6 pmol/24 h, and did not correlate with body weight, age, or plasma vitamin C concentration. The 5 smokers excreted 30.5 +/- 8.5 and the 28 nonsmokers excreted 38.6 +/- 2.4 pmol epsilondA per 24 h, p = .37 (mean SEM). The demonstrated level of performance suggests the future applicability of this method to studies of cancer and other diseases related to oxidative stress in humans. (C) 2004 Elsevier Inc. All rights reserved.
Original languageEnglish
JournalFree Radical Biology and Medicine
Issue number11
Pages (from-to)1383-1392
Publication statusPublished - 2004
Externally publishedYes


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