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Preparation of Nucleotide Advanced Glycation Endproducts—Imidazopurinone Adducts Formed by Glycation of Deoxyguanosine with Glyoxal and Methylglyoxal
Author(s) -
Fleming Thomas,
Rabbani Naila,
Thornalley Paul J.
Publication year - 2008
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1196/annals.1433.037
Subject(s) - methylglyoxal , chemistry , deoxyguanosine , glycation , glyoxal , nucleotide , biochemistry , isotope dilution , adduct , dna , chromatography , tandem mass spectrometry , nuclease , mass spectrometry , organic chemistry , enzyme , receptor , gene
An analytical procedure was developed for nucleotide advanced glycation endproducts formed by the reaction of glyoxal and methylglyoxal with deoxyguanosine under physiological conditions. For this, the imidazopurinone derivatives, 3‐(2′‐deoxyribosyl)‐6,7‐dihydro‐6,7‐dihydroxyimidazo[2,3‐b]purin‐9(8)one (dG‐G) and 3‐(2′‐deoxyribosyl)‐6,7‐dihydro‐6,7‐dihydroxy‐6‐methylimidazo‐[2,3‐b]purine‐9(8)one (dG‐MG), were prepared. Authentic standard and stable isotope‐substituted standard adducts were prepared and an isotopic dilution analysis assay methodology was developed using liquid chromatography with tandem mass spectrometry and optimized DNA extraction and nuclease digestion procedures. Analysis of dG‐G, dG‐MG, and the oxidative marker 8‐hydroxydeoxyguanosine in the DNA of cultured human cells and mononuclear leukocytes showed that nucleotide advanced glycation endproducts are major markers of DNA damage in human cells.