Accurate and precise quantification of Cu,Zn-SOD in human red blood cells using species-specific double and triple IDMS

Julia Gleitzmann, Andrea Raab, Dirk Schulze, Hermann Waetzig, Jorg Feldmann, Claudia Swart

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Abstract

Cu,Zn-superoxide dismutase (SOD1) is a protein involved in the antioxidant defense system of the body responsible for the dismutation of the superoxide anion. It contains two Cu and two Zn ions per molecule. As this protein is also involved in several diseases it is used in clinical diagnostics as biomarker, which requires the accurate and reliable determination of SOD1. Therefore, a candidate reference measurement procedure for the quantification of this protein in human erythrocytes was developed using species-specific isotope dilution mass spectrometry (IDMS), a method giving results traceable to the International System of Units (SI). The measurement procedure was validated with regard to a metrological point of view. Commercially available SOD1 was thoroughly characterized to be used as pure protein calibration standard in the IDMS approaches. Furthermore, 65Cu and 67Zn labeled SOD was produced to be used as spike material required for species-specific IDMS. Finally, SOD1 was quantified in human erythrocytes using both double and triple IDMS and a complete uncertainty budget for both approaches was estimated according to the Guide for the Expression of Uncertainty in Measurement (GUM). A calculated mass fraction of SOD1 with its associated expanded uncertainty of (63.94 ± 0.93) μg/g (n = 30) for double and (64.02 ± 0.96) μg/g (n = 30) for triple IDMS was obtained.
Original languageEnglish
Pages (from-to)1922-1928
Number of pages7
JournalJournal of Analytical Atomic Spectrometry
Volume31
Issue number9
Early online date17 Mar 2016
DOIs
Publication statusPublished - 1 Sep 2016

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Isotopes
Dilution
Mass spectrometry
Blood
Cells
Proteins
Metric system
Biomarkers
Superoxides
Antioxidants
Calibration
Ions
Molecules
Uncertainty

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Accurate and precise quantification of Cu,Zn-SOD in human red blood cells using species-specific double and triple IDMS. / Gleitzmann, Julia; Raab, Andrea; Schulze, Dirk; Waetzig, Hermann; Feldmann, Jorg; Swart, Claudia.

In: Journal of Analytical Atomic Spectrometry, Vol. 31, No. 9, 01.09.2016, p. 1922-1928.

Research output: Contribution to journalArticle

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