Guarnieri C, Muscari C, Stefanelli C, Giaccari A, Zini M, Di Biase S
Department of Biochemistry G. Moruzzi, University of Bologna, Italy.
J Chromatogr B Biomed Appl. 1994 Jun 3;656(1):209-13. doi: 10.1016/0378-4347(94)00044-1.
8-Hydroxydeoxyguanosine (8-OH-dG) is widely recognized as a marker of DNA oxidation. Until now, 8-OH-dG has been measured by high-performance liquid chromatography or by gas chromatography-mass spectrometry. A method is reported that detects oxidative derivatives of deoxynucleosides by micellar electrokinetic capillary chromatography. Single-stranded DNA was incubated in the presence of 50 mM hydrogen peroxide-10 mM ascorbic acid and hydrolysed by enzymatic digestion. The order of electrophoretic mobilities of deoxynucleosides was dC > dA > T > dG > 8-OH-dG. 8-OH-dG was determined by introducing a laboratory-prepared internal standard. Two additional major oxidative derivatives were identified by comparing the electropherogram of the oxidized DNA with that of the oxidized standard deoxyguanosine.
8-羟基脱氧鸟苷(8-OH-dG)被广泛认为是DNA氧化的标志物。到目前为止,8-OH-dG已通过高效液相色谱法或气相色谱-质谱法进行测定。本文报道了一种通过胶束电动毛细管色谱法检测脱氧核苷氧化衍生物的方法。将单链DNA在50 mM过氧化氢-10 mM抗坏血酸存在下孵育,并通过酶消化进行水解。脱氧核苷的电泳迁移率顺序为dC > dA > T > dG > 8-OH-dG。通过引入实验室制备的内标物来测定8-OH-dG。通过将氧化DNA的电泳图与氧化标准脱氧鸟苷的电泳图进行比较,鉴定出另外两种主要的氧化衍生物。