[Detection of 8-hydroxyguanine in the DNA of the H2O2-treated HL-60 cells by GC/FID].
This paper describes the DNA damage by hydrogen peroxide in HL-60 cells and the detection of 8-hydroxyguanine produced. The number of cells was at least about 2 x 10(7)/sample. Treatment of HL-60 cells with 0.4 mmol/L hydrogen peroxide for 24 h would result in DNA damage. The DNA samples were isolated, then qualitatively and quantitatively analyzed before hydrolysis. The results of experiments showed that 100-300 microL 98% formic acid could hydrolyze about 0.12 mg DNA samples completely in 150 degrees C for 1 h. After lyophilizing the DNA samples for 6-8 h, one DNA sample was trimethylsilylated completely with 150 microL BSTFA in 150 degrees C for 1 h. The recovery of trimethylsilylated 8-hydroxyguanine was 81.70%. The trimethylsilylated 8-hydroxyguanine was identified by GC/FID and CGC/MS-SIM parallely.
Zhang
H
H
Institute of Medical Biochemistry, Guangdong Medical College, Zhanjiang, 524023.
Zhu
Q
Q
Mo
L
L
Zhuang
H
H
Cai
C
C
D002849:Chromatography, Gas; D004247:DNA / Q000032:analysis / Q000737:chemistry*; D006147:Guanine / Q000031:analogs & derivatives* / Q000032:analysis; D018922:HL-60 Cells; D006801:Humans; D006859:Hydrogen; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D015203:Reproducibility of Results
C024829:8-hydroxyguanine; D006147:Guanine; D006859:Hydrogen; D004247:DNA; D006861:Hydrogen Peroxide