赵腾雯, 李海斌, 张思雨, 周晓滢, 王馨蒙, 崔师伟. 超高效液相色谱-串联质谱测定尿中8-羟基脱氧鸟苷[J]. 职业卫生与应急救援, 2025, 43(2): 259-264. DOI: 10.16369/j.oher.issn.1007-1326.2025.240429
引用本文: 赵腾雯, 李海斌, 张思雨, 周晓滢, 王馨蒙, 崔师伟. 超高效液相色谱-串联质谱测定尿中8-羟基脱氧鸟苷[J]. 职业卫生与应急救援, 2025, 43(2): 259-264. DOI: 10.16369/j.oher.issn.1007-1326.2025.240429
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ZHAO Tengwen, LI Haibin, ZHANG Siyu, ZHOU Xiaoying, WANG Xinmeng, CUI Shiwei. Determination of 8-hydroxy-2'-deoxyguanosine in urine by UHPLC-MS/MS[J]. Occupational Health and Emergency Rescue, 2025, 43(2): 259-264. DOI: 10.16369/j.oher.issn.1007-1326.2025.240429
Citation: ZHAO Tengwen, LI Haibin, ZHANG Siyu, ZHOU Xiaoying, WANG Xinmeng, CUI Shiwei. Determination of 8-hydroxy-2'-deoxyguanosine in urine by UHPLC-MS/MS[J]. Occupational Health and Emergency Rescue, 2025, 43(2): 259-264. DOI: 10.16369/j.oher.issn.1007-1326.2025.240429
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超高效液相色谱-串联质谱测定尿中8-羟基脱氧鸟苷

Determination of 8-hydroxy-2'-deoxyguanosine in urine by UHPLC-MS/MS

    摘要
  • 摘要:
    目的 建立尿中8-羟基脱氧鸟苷的超高效液相色谱-串联质谱检测方法，为DNA氧化损伤的定量评估提供方法支持。
    方法 尿液样品经离心后除去沉淀物，所得上清液利用甲酸-甲酸铵缓冲液稀释，使用亲水亲油平衡（HLB）固相萃取柱对其净化，随后经BEH C18色谱柱分离，三重四极杆质谱进行检测；使用多反应监测模式及同位素内标法进行定量分析。
    结果 尿液中8-羟基脱氧鸟苷在0.5~20 μg/L范围内线性关系良好，该方法的检出限和定量限分别为0.06 μg/L和0.20 μg/L，加标回收率为98.1%~98.4%，批内精密度、批间精密度（n=6）分别为1.00%~2.35%、1.23%~3.93%。
    结论 该方法具有良好的灵敏度和准确度，适用于尿液样品中8-羟基脱氧鸟苷的测定。

     

    Abstract:
    Objective To establish a ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/ MS) method for the detection of 8-hydroxy-2'-deoxyguanosine in urine, providing methodological support for the quantitative assessment of DNA oxidative damage.
    Methods Urine samples were centrifuged to remove precipitates, and the supernatant was diluted with formic acid-ammonium formate buffer. The samples were purified using hydrophilic-lipophilic balance (HLB) solid-phase extraction columns, separated via a BEH C18 chromatography column, and detected by triple quadrupole mass spectrometry. Quantitative analysis was conducted using multiple reaction monitoring (MRM) mode combined with an isotope internal standard method.
    Results The detection method demonstrated good linearity in the range of 0.5 to 20 μg/L for 8-hydroxy-2'-deoxyguanosine in urine. The limits of detection (LOD) and quantitation (LOQ) were 0.06 μg/L and 0.20 μg/L, respectively. The spiked recovery rates ranged from 98.1% to 98.4%. Intra-batch precision and inter-batch precision (n = 6) were 1.00% to 2.35% and 1.23% to 3.93%, respectively.
    Conclusions This method showed high sensitivity and accuracy, making it suitable for the determination of 8-hydroxy-2'-deoxyguanosine in urine samples.

     

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