Determination of 8-OHGua by LC-MS/MS after Acid Hydrolysis of Oxidative Damaged DNA
Küçük Resim Yok
Tarih
2023
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Iranian Chemical Soc
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
A variety of influences can cause DNA damage to the genome. The hydroxyl radical attacks the C-8 atom of guanine, forming 8-hydroxyguanine (8-OHGua) or 8-hydroxy-2'-deoxyguanosine (8-OH-2'dG) which are important indicators of oxidative damage on DNA. Determining these damaged base products can be accomplished through measurement by LC-MS/MS after enzymatic hydrolysis or measurement by GC-MS/MS after chemical hydrolysis. In this study, it was aimed to hydrolyze DNA using various strong acids and to measure 8-hydroxyguanine by LC-MS/MS. In the first stage of the study, the nucleoside 8- hydroxy-2'-deoxyguanosine was treated with HCl (2 M and 6 M), TCA (10%), TFA (10%), o-phosphoric acid (2 M), methanesulfonic acid (2 M), and formic acid (80%). The amounts of 8-hydroxyguanine were determined by LC-MS/MS. It has been identified that formic acid with the highest yield (70%) hydrolyzes the beta-glycosidic linkage between the base and the sugar. Subsequently, oxidative damage was induced on calf thymus DNA by producing hydroxyl radicals via the Fenton reaction. The resulting oxidative damaged DNA was hydrolyzed using formic acid. The amount of 8-hydroxyguanine was then determined using LC-MS/MS. Based on the results obtained, it was observed that the acidic hydrolysis applied was effective in breaking the N-glycosidic bond, but not effective in breaking the phosphodiester bond of oxidatively damaged DNA.
Açıklama
Anahtar Kelimeler
8-OHG, 8-OH-2'dg, LC-MS/MS, Acidic Hydrolysis, Thymus DNA, N-Glycosidic Bond, Mass-Spectrometry, Repair, Base, 8-Oxo-7,8-Dihydroguanine, 8-Hydroxydeoxyguanosine, 8-Oxoguanine, Biomarkers, Protection, Cells
Kaynak
Analytical And Bioanalytical Chemistry Research
WoS Q Değeri
N/A
Scopus Q Değeri
Q3
Cilt
10
Sayı
4
