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A fully validated bioanalytical method using an UHPLC–MS/MS system for quantification of DNA and RNA oxidative stress biomarkers

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Abstract

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.

UHPLC-MS/MS analysis of DNA and RNA oxidative stress biomarkers

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Acknowledgements

This work was supported by projects SVV 260 412, MH CZ - DRO (UHHK, 00179906) and Progress Q42.

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Authors and Affiliations

  1. Department of Analytical Chemistry, Faculty of Pharmacy, Charles University, Akademika Heyrovskeho 1203, 50005, Hradec Kralove, Czech Republic

    Barbora Cervinkova, Lenka Kujovska Krcmova, Veronika Sestakova & Petr Solich

  2. 3rd Internal Gerontometabolic Clinic, University Hospital, Sokolska 581, 50005, Hradec Kralove, Czech Republic

    Barbora Cervinkova, Lenka Kujovska Krcmova & Dagmar Solichova

Authors
  1. Barbora Cervinkova
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  2. Lenka Kujovska Krcmova
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  3. Veronika Sestakova
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  4. Dagmar Solichova
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  5. Petr Solich
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Corresponding author

Correspondence to Lenka Kujovska Krcmova.

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This study has been approved by the University Hospital Hradec Kralove ethics committee 201606 S21P and has been performed in accordance with the ethical standards.

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The authors declare that they have no conflict of interest.

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Cervinkova, B., Krcmova, L.K., Sestakova, V. et al. A fully validated bioanalytical method using an UHPLC–MS/MS system for quantification of DNA and RNA oxidative stress biomarkers. Anal Bioanal Chem 409, 3611–3621 (2017). https://doi.org/10.1007/s00216-017-0301-2

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  • DOI: https://doi.org/10.1007/s00216-017-0301-2

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