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目的对利用高效液相色谱-电化学法(HPLC-ECD)测定HepG2细胞内8-羟基脱氧鸟苷(8-OH-dG)的方法进行研究。方法 HepG2细胞经前处理后,利用美国Varian Prostar高效液相色谱仪电化学检测器检测8-OH-dG水平。结果 8-OH-dG峰与杂质峰分离良好,8-OH-dG浓度在1～25 ng/ml之间呈良好线性,流动相最佳pH值为5.2,最佳工作电压为+0.60V,最低检测浓度为0.39 ng/ml,在线性范围内的3种质量浓度20、5、1 ng/ml下进行的测定,8-OH-dG回收率分别为(86.1+1.2)%、(85.3+2.3)%、(84.1+1.8)%。精密度试验的批内RSD在2.8%～6.7%,精密度试验的批间RSD在6.5%～8.4%。结论该法检测HepG2细胞内8-OH-dG灵敏、准确、重复性好,操作简便,值得推广。
Objective To study the method for the determination of 8-OH-dG in HepG2 cells by high performance liquid chromatography-electrochemical method (HPLC-ECD). Methods HepG2 cells were pre-treated and the level of 8-OH-dG was detected by a Varian Prostar high performance liquid chromatography electrochemical detector. Results The 8-OH-dG peak was well separated from the impurity peak. The linearity of 8-OH-dG ranged from 1 ng / mL to 25 ng / mL. The optimum pH of mobile phase was 5.2 and the optimal working voltage was + 0.60 V. The minimum detectable concentration was 0.39 ng / ml. The linear range of the three concentrations of 20, 5, 1 ng / ml were determined. The recoveries of 8-OH-dG were (86.1 + 1.2) 2.3)%, (84.1 + 1.8)%. The intra-assay RSD of the precision test ranged from 2.8% to 6.7% and the inter-assay RSD of the precision test ranged from 6.5% to 8.4%. Conclusion The method of detecting HepG2 cells 8-OH-dG sensitive, accurate, reproducible, easy to operate, it is worth promoting.