A fully validated bioanalytical method using an UHPLC-MS/MS system for quantification of DNA and RNA oxidative stress biomarkers.
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 ([15N]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. Graphical abstract UHPLC-MS/MS analysis of DNA and RNA oxidative stress biomarkers.
Department of Analytical Chemistry, Faculty of Pharmacy, Charles University, Akademika Heyrovskeho 1203, 50005, Hradec Kralove, Czech Republic.|3rd Internal Gerontometabolic Clinic, University Hospital, Sokolska 581, 50005, Hradec Kralove, Czech Republic.
Barbora
B
Cervinkova
Department of Analytical Chemistry, Faculty of Pharmacy, Charles University, Akademika Heyrovskeho 1203, 50005, Hradec Kralove, Czech Republic. LenkaKrcmova@seznam.cz.|3rd Internal Gerontometabolic Clinic, University Hospital, Sokolska 581, 50005, Hradec Kralove, Czech Republic. LenkaKrcmova@seznam.cz.
Lenka Kujovska
LK
Krcmova
Department of Analytical Chemistry, Faculty of Pharmacy, Charles University, Akademika Heyrovskeho 1203, 50005, Hradec Kralove, Czech Republic.
Veronika
V
Sestakova
3rd Internal Gerontometabolic Clinic, University Hospital, Sokolska 581, 50005, Hradec Kralove, Czech Republic.
Dagmar
D
Solichova
Department of Analytical Chemistry, Faculty of Pharmacy, Charles University, Akademika Heyrovskeho 1203, 50005, Hradec Kralove, Czech Republic.
Petr
P
Solich
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000328:Adult; D015415:Biomarkers / Q000652:urine; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003401:Creatine / Q000652:urine*; D004247:DNA / Q000652:urine; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000652:urine; D005260:Female; D006151:Guanosine / Q000031:analogs & derivatives* / Q000652:urine; D006801:Humans; D057230:Limit of Detection; D008297:Male; D009369:Neoplasms / Q000652:urine; D019636:Neurodegenerative Diseases / Q000652:urine; D018384:Oxidative Stress; D012313:RNA / Q000652:urine; D052616:Solid Phase Extraction / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*; D055815:Young Adult
D015415:Biomarkers; D006151:Guanosine; C046215:8-hydroxyguanosine; D012313:RNA; D000080242:8-Hydroxy-2'-Deoxyguanosine; D004247:DNA; D003849:Deoxyguanosine; D003401:Creatine
8-Hydroxy-2-deoxyguanosine; 8-Hydroxyguanosine; Cancer; Creatinine; Human urine; UHPLC–MS/MS