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

Detection of 8-Hydroxyguanine in the DNA of the H_2O_2-Treated HL-60 Cells by GC/FID

用0．4mmol／LH2O2处理HL－60细胞株24h，采用气相色谱／氢火焰检测器（GC／FID）检测DNA氧化损伤产物8-羟基鸟嘌呤，并用气相色谱质谱仪选择性离子检测（CGC／MS－SIM）对其进一步鉴定。所用方法的平均回收率为81．7％，RSD小于5％。

This paper describes the DNA damage by hydrogen peroxide in HL-60 cells and the detection of 8hydroxyguanine produced. The number of cells was at least about 2× 107/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 μL 98% formic acid could hydrolyze about 0. 12 mg DNA samples completely in 150℃ for 1h. After lyophilizing the DNA samples for 6-8 h,one DNA sample was trimethylsilylated completely with 150μL BSTFA in 150℃ 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.