Journal
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 409, Issue 14, Pages 3611-3621Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-017-0301-2
Keywords
8-Hydroxy-2-deoxyguanosine; 8-Hydroxyguanosine; Creatinine; Cancer; UHPLC-MS/MS; Human urine
Funding
- MH CZ - DRO (UHHK) [00179906]
- Progress Q42
- [SVV 260 412]
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A new, rapid and effective ultra-high-performance liquid chromatography method with mass spectrometry detection is described for the separation and quantification of 8-hydroxy-2-deoxyguanosine, 8-hydroxyguanosine and creatinine in human urine. The present study uses an isotope-labelled internal standard ([N-15](5)-8-hydroxy-2-deoxyguanosine), a BIO core-shell stationary phase and an isocratic elution of methanol and water. Sample preparation of human urine was performed by solid-phase extraction (SPE) on Oasis HLB cartridges with methanol/water 50: 50 (v/v) elution. Extraction recoveries ranged from 98.1% to 109.2%. Biological extracts showed high short-term stability. Several aspects of this procedure make it suitable for both clinical and research purposes: a short elution time of less than 3.2 min, an intra-day precision of 2.5-8.9%, an inter-day precision of 3.4-8.7% and low limits of quantification (27.7 nM for 8-hydroxyguanosine, 6.0 nM for 8-hydroxy-2-deoxyguanosine). Finally, simultaneous analysis of DNA and RNA oxidative stress biomarkers is a useful tool for monitoring disease progression in neurodegenerative disorders and cancer.
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