Development of a quality control material for the measurement of 8-oxo-7,8-dihydro-2'-deoxyguanosine, an in vivo marker of oxidative stress, and comparison of results from different laboratories.
The measurement of 8-oxo-7,8-dihydro-2'-deoxyguanosine is an increasingly popular marker of in vivo oxidative damage to DNA. A random-sequence 21-mer oligonucleotide 5'-TCA GXC GTA CGT GAT CTC AGT-3' in which X was 8-oxo-guanine (8-oxo-G) was purified and accurate determination of the oxidised base was confirmed by a 32P-end labelling strategy. The lyophilised material was analysed for its absolute content of 8-oxo-dG by several major laboratories in Europe and one in Japan. Most laboratories using HPLC-ECD underestimated, while GC-MS-SIM overestimated the level of the lesion. HPLC-ECD measured the target value with greatest accuracy. The results also suggest that none of the procedures can accurately quantitate levels of 1 in 10(6) 8-oxo-(d)G in DNA.
Lunec
J
J
MRC Centre for Mechanisms in Human Toxicity, Leicester, UK. jl20@le.ac.uk
Herbert
K E
KE
Jones GDD
Dickinson
L
L
Evans
M
M
Mistry
N
N
Mistry
P
P
Chauhan
D
D
Capper
G
G
Zheng
Q
Q
D015415:Biomarkers / Q000032:analysis*; D002853:Chromatography, Liquid; D019411:Clinical Laboratory Techniques; D003848:Deoxyguanine Nucleotides / Q000032:analysis*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D009841:Oligonucleotides / Q000737:chemistry; D018384:Oxidative Stress; D011786:Quality Control; D017422:Sequence Analysis, DNA; D013056:Spectrophotometry, Ultraviolet
D015415:Biomarkers; D003848:Deoxyguanine Nucleotides; D009841:Oligonucleotides; C078206:8-oxodeoxyguanosine triphosphate