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

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.