HepG2细胞DNA损伤中8-OH-dG含量测定的高效液相色谱-电化学法  

Determination of 8-OH-dG Level in HepG2 Cell with DNA Damage by HPLC-ECD

作　　者：鲁文红[1] 来瑞平[1] 张裕曾[1] 

机构地区：[1]华中科技大学同济医学院公共卫生学院环境与健康教育部重点实验室,湖北省武汉市430030

出　　处：《职业与健康》2011年第19期2197-2199,共3页Occupation and Health

基　　金：国家自然科学基金项目(项目编号:30471500)

摘　　要：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 investigate the methodology of high-performance liquid chromatography with electrochemical detector（HPLC-ECD）in damage DNA 8-hydroxydeoxyguanosine（8-OH-dG） quantification of HepG2 cell. [Methods]After pre-processing of HepG2 cell,quantification of damage DNA 8-OH-dG was acquired with HPLC-ECD of American Varian Prostar. [Results]It was a large separation between the 8-OH-dG peak and foreign matter peak and the calibration curves of 8-OH-dG were linear range from 1 to 25 ng/ml.The best PH of the mobile phase and working voltage were 5.2 and ＋0.60 v respectively.0.39 ng/ml was the lowest concentration to be measured.The method recoveries were（86.1＋1.2）%,（85.3＋2.3）%,（84.1＋1.8）% at the concentration of 20,5,1ng/ml in linear range.The within-batch RSD of precision test was 2.8%~6.7%.and the between-batch RSD was 6.5%~8.4%. [Conclusion]The method was found to be sensitive and accurate for the determination of 8-OH-dG in damage DNA of HepG2 cell and was deserve to promote.

关 键 词：8-羟基脱氧鸟苷 高效液相色谱-电化学 

分 类 号：R972[医药卫生—药品]