Micellar electrokinetic capillary chromatography of 8-hydroxydeoxyguanosine and other oxidized derivatives of DNA.
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.
Department of Biochemistry G. Moruzzi, University of Bologna, Italy.
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C
Guarnieri
C
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Muscari
C
C
Stefanelli
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A
Giaccari
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M
Zini
S
S
Di Biase
D000080242:8-Hydroxy-2'-Deoxyguanosine; D002845:Chromatography; D018736:DNA Adducts / Q000737:chemistry / Q000302:isolation & purification*; D004277:DNA, Single-Stranded / Q000737:chemistry; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000302:isolation & purification; D004563:Electrochemistry; D004586:Electrophoresis; D008823:Micelles; D009691:Nucleic Acid Denaturation; D010084:Oxidation-Reduction; D013056:Spectrophotometry, Ultraviolet
D018736:DNA Adducts; D004277:DNA, Single-Stranded; D008823:Micelles; D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine