Detection of 8-Hydroxyguanine in the DNA of the H_2O_2-Treated HL-60 Cells by GC/FID
气相色谱/氢火焰检测器检测HL-60细胞DNA中氧化损伤产物8-羟基鸟嘌呤的研究

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.