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Title: A novel 14C-postlabeling assay using accelerator mass spectrometry for the detection of O6-methyldeoxy-guanosine adducts
Authors: Tompkins, Elaine M.
Farmer, Peter B.
Lamb, John H.
Jukes, Rebekah
Dingley, Karen
Ubick, Esther
Turteltaub, Kenneth W.
Martin, Elizabeth A.
Brown, Karen
First Published: 8-Feb-2006
Publisher: John Wiley & Sons
Citation: Rapid Communications in Mass Spectrometry, 2006, 20 (5), pp. 883-891.
Abstract: Accelerator mass spectrometry (AMS) is currently one of the most sensitive methods available for the trace detection of DNA adducts and is particularly valuable for measuring adducts in humans or animal models. However, the standard approach requires administration of a radiolabeled compound. As an alternative, we have developed a preliminary 14C-postlabeling assay for detection of the highly mutagenic O6-methyldeoxyguanosine (O6-MedG), by AMS. Procedures were developed for derivatising O6-MedG using unlabeled acetic anhydride. Using conventional liquid chromatography/mass spectrometry (LC/MS) analysis, the limit of detection (LOD) for the major product, triacetylated O6-MedG, was 10 fmol. On reaction of O6-MedG with 14C-acetic anhydride, using a specially designed enclosed system, the predominant product was 14C-di-acetyl O6-MedG. This change in reaction profile was due to a modification of the reaction procedure, introduced as a necessary safety precaution. The LOD for 14C-di-acetyl O6-MedG by AMS was determined as 79 amol, ∼18 000-fold lower than that achievable by liquid scintillation counting (LSC). Although the assay has so far only been carried out with labeled standards, the degree of sensitivity obtained illustrates the potential of this assay for measuring O6-MedG levels in humans.
ISSN: 0951-4198
Type: Article
Description: This paper was published as Rapid Communications in Mass Spectrometry, 2006, 20 (5), pp. 883-891. It is available from DOI: 10.1002/rcm.2370
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Appears in Collections:Published Articles, Dept. of Cancer Studies and Molecular Medicine

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