Assessment of oxidative damage to proteins and DNA in urine of newborn infants by a validated UPLC-MS/MS approach

Julia Kuligowski; Isabel Torres-Cuevas; Guillermo Quintás; Denise Rook; Johannes B van Goudoever; Elena Cubells; Miguel Asensi; Isabel Lliso; Antonio Nuñez; Máximo Vento; Javier Escobar

doi:10.1371/journal.pone.0093703

Assessment of oxidative damage to proteins and DNA in urine of newborn infants by a validated UPLC-MS/MS approach

PLoS One. 2014 Apr 2;9(4):e93703. doi: 10.1371/journal.pone.0093703. eCollection 2014.

Affiliations

¹ Neonatal Research Group, Health Research Institute Hospital La Fe, Valencia, Spain.
² Leitat Technological Center, BioInVitro Division, Valencia, Spain.
³ Department of Pediatrics, Division of Neonatology, Erasmus Medical Centre-Sophia Children's Hospital, Rotterdam, The Netherlands.
⁴ Department of Pediatrics, Division of Neonatology, Erasmus Medical Centre-Sophia Children's Hospital, Rotterdam, The Netherlands; Department of Pediatrics, Free University Medical Centre Amsterdam, Amsterdam, The Netherlands; Department of Pediatrics, Emma Children's Hospital, Academic Medical Center, Amsterdam, The Netherlands.
⁵ Department of Physiology, University of Valencia, Burjassot, Valencia, Spain.
⁶ Neonatal Research Group, Health Research Institute Hospital La Fe, Valencia, Spain; Division of Neonatology, University & Polytechnic Hospital La Fe, Valencia, Spain.

Abstract

The assessment of oxidative stress is highly relevant in clinical Perinatology as it is associated to adverse outcomes in newborn infants. This study summarizes results from the validation of an Ultra Performance Liquid Chromatography-tandem Mass Spectrometry (UPLC-MS/MS) method for the simultaneous quantification of the urinary concentrations of a set of endogenous biomarkers, capable to provide a valid snapshot of the oxidative stress status applicable in human clinical trials, especially in the field of Perinatology. The set of analytes included are phenylalanine (Phe), para-tyrosine (p-Tyr), ortho-tyrosine (o-Tyr), meta-tyrosine (m-Tyr), 3-NO2-tyrosine (3NO2-Tyr), 3-Cl-tyrosine (3Cl-Tyr), 2'-deoxyguanosine (2dG) and 8-hydroxy-2'-deoxyguanosine (8OHdG). Following the FDA-based guidelines, appropriate levels of accuracy and precision, as well as adequate levels of sensitivity with limits of detection (LODs) in the low nanomolar (nmol/L) range were confirmed after method validation. The validity of the proposed UPLC-MS/MS method was assessed by analysing urine samples from a clinical trial in extremely low birth weight (ELBW) infants randomized to be resuscitated with two different initial inspiratory fractions of oxygen.

Publication types

Research Support, Non-U.S. Gov't
Validation Study

MeSH terms

DNA / urine*
Humans
Infant, Newborn
Oxidative Stress*
Tandem Mass Spectrometry / methods*

Substances

DNA

Grants and funding

Javier Escobar acknowledges his personal grant Sara Borrell CD11/00154 from the Instituto Carlos III (Ministry of Economy and Competitiveness); Julia Kuligowski acknowledges her personal grant Sara Borrell CD12/00667 from the Instituto Carlos III (Ministry of Economy and Competitiveness); Isabel Torres-Cuevas acknowledges her personal grant PFIS FI12/00109 from the Instituto Carlos III (Ministry of Economy and Competitiveness); Máximo Vento acknowledges FIS PI11/0313 grant from the Instituto Carlos III (Ministry of Economy and Competitiveness) and EC11-244 from the Spanish Ministry of Health, Social Services and Equality; and Guillermo Quintás acknowledges financial support from the Ministerio de Economia y Competitividad(SAF2012-39948). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.