超高效液相色谱串联质谱法测定人尿中的8-羟基脱氧鸟苷

Determination of 8-OHdG in human urine by ultra-high performance liquid chromatography tandem mass spectrometry

目的建立一种超高效液相色谱-串联质谱法(UPLC-MS/MS)简单、快速的分析人尿液中8-羟基脱氧鸟苷(8-OHd G)含量的检测方法。方法人尿样品经离心后,通过HLB固相萃取柱进行净化处理,采用5%的甲醇-水(含0.1%甲酸)作为色谱的分离条件,在MRM模式下采集8-OHd G(m/z=284.2>168.2及m/z=284.2>140.2)的信号,用外标法定量。结果本文报道的检测方法具有较高的灵敏度,8-OHd G在线性范围0.1 ng/ml^2.0 ng/ml内线性关系良好,相关系数为0.9995,定量下限为0.02 ng/ml,低、中、高3个加标水平的回收率为90.2%~103.7%,相对标准偏差为1.1%~6.8%。结论该方法操作简便,与前人报道的检测结果一致,也可以应用于评价各种氧化因素引起的DNA氧化损伤。

Objective This paper established a simple,rapid method for the analysis of 8-OHd G in human urine with liquid chromatography-tandem mass spectrometry（ UPLC-MS / MS）. Methods After centrifugation of human urine,HLB solid phase extraction column was used for purification. Then 5% methanol-water（ containing 0. 1% formic acid） as chromatographic separation conditions and in the MRM mode 8-OHd G（ m / z = 284. 2〉 168. 2 and m / z = 284. 2 140. 2） signals were gained The quantification was accomplished by external standard method. Results The detection method has high sensitivity,8-OHd G has a good linear relationship in the linear range（ 0. 1 ng / ml - 2. 0 ng / ml）,the correlation coefficient was 0. 9995,the lower limit of quantification of 0. 02 ng / ml,low,medium and high levels spiked recoveries of 90. 2% - 103. 7%,and the relative standard deviation was 1. 1% - 6. 8%. Conclusion The method is simple and the test results are consistent with the previous reports,which can also be applied to the evaluation of oxidative DNA damage caused by various oxidation factors.