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BMC Pediatrics
Published in: BMC Pediatrics 1/2017

Open Access 01-12-2017 | Research article

Reduced glutathione and glutathione disulfide in the blood of glucose-6-phosphate dehydrogenase-deficient newborns

Authors: Zhen-hua Gong, Guo-li Tian, Qi-wei Huang, Yan-min Wang, Hong-ping Xu

Published in: BMC Pediatrics | Issue 1/2017

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Abstract

Background

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is commonly detected during mass screening for neonatal disease. We developed a method to measure reduced glutathione (GSH) and glutathione disulfide (GSSG) using tandem mass spectrometry (MS/MS) for detecting G6PD deficiency.

Methods

The concentration of GSH and the GSH/GSSG ratio in newborn dry-blood-spot (DBS) screening and in blood plus sodium citrate for test confirmation were examined by MS/MS using labeled glycine as an internal standard.

Results

G6PD-deficient newborns had a lower GSH content (242.9 ± 15.9 μmol/L)and GSH/GSSG ratio (14.9 ± 7.2) than neonatal controls (370.0 ± 53.2 μmol/L and 46.7 ± 19.6, respectively). Although the results showed a significance of P < 0.001 for DBS samples plus sodium citrate that were examined the first day after preparation, there were no significant differences in the mean GSH concentration and GSH/GSSG ratio between the G6PD deficiency-positive and negative groups when examined three days after sample preparation.

Conclusion

The concentration of GSH and the ratio of GSH/GSSG in blood measured using MS/MS on the first day of sample preparation are consistent with G6PD activity and are helpful for diagnosing G6PD deficiency.
Literature
1.
Cappellini MD, Fiorelli G. Glucose-6-phosphate dehydrogenase deficiency. Lancet. 2008;371(9606):64–74.CrossRefPubMed
2.
Arese P, Gallo V, Pantaleo A, Turrini F. Life and death of glucose-6-phosphate dehydrogenase (G6PD) deficient erythrocytes - role of redox stress and band 3 modifications. Transfus Med Hemother. 2012;39(5):328–34.CrossRefPubMedPubMedCentral
3.
van Zwieten R, Verhoeven AJ, Roos D. Inborn defects in the antioxidant systems of human red blood cells. Free Radic Biol Med. 2014;67:377–86.
4.
Mason PJ, Bautista JM, Gilsanz F. G6PD deficiency: The genotype-phenotype association. Blood rev. 2007;21(5):267–83.
5.
Shimo H, Nishino T, Tomita M. Predicting the kinetic properties associated with redox imbalance after oxidative crisis in G6PD-deficient erythrocytes: a simulation study. Adv Hematol. 2011;2011:398945.CrossRefPubMedPubMedCentral
6.
Winterbourn CC. Reconciling the chemistry and biology of reactive oxygen species. Nat Chem Biol. 2008;4(5):278–86.CrossRefPubMed
7.
Garibotto G, Sofia A, Saffioti S, et al. Interorgan exchange of aminothiols in humans. Am J Physiol Endocrinol Metab. 2003;284(4):E757–63.CrossRefPubMed
8.
Dass PD, Bermes EW Jr, Holmes EW. Renal and hepatic output of glutathione in plasma and whole blood. Biochim Biophys Acta. 1992;1156(1):99–102.CrossRefPubMed
9.
Giustarini D, Milzani A, Dalle-Donne I, Rossi R. Red blood cells as a physiological source of glutathione for extracellular fluids. Blood Cells Mol Dis. 2008;40(2):174–9.CrossRefPubMed
10.
Kalinina EV, Chernov NN, Novichkova MD. Role of glutathione, glutathione transferase, and glutaredoxin in regulation of redox-dependent processes. Biochemistry (Mosc). 2014;79(13):1562–83.CrossRef
11.
Rossi R, Milzani A, Dalle-Donne I, et al. Different metabolizing ability of thiol reactants in human and rat blood: biochemical and pharmacological implications. J Biol Chem. 2001;276(10):7004–10.CrossRefPubMed
12.
Cakir Y, Yildiz D. Efflux of glutathione and glutathione complexes from human erythrocytes in response to vanadate. Blood Cells Mol Dis. 2013;50(1):1–7.CrossRefPubMed
13.
Ellison I, Richie JP Jr. Mechanisms of glutathione disulfide efflux from erythrocytes. Biochem Pharmacol. 2012;83(1):164–9.CrossRefPubMed
14.
Sakhi AK, Blomhoff R, Gundersen TE. Simultaneous and trace determination of reduced and oxidizedglutathione in minute plasma samples using dual mode fluorescence detection and column switching high performance liquid chromatography. J Chromatogr A. 2007;1142(2):1.CrossRef
15.
Rellán-Alvarez R, Hernández LE, Abadía J, Alvarez-Fernández A. Direct and simultaneous determination of reduced and oxidized glutathione and homoglutathione by liquid chromatography-electrospray/mass spectrometry in plant tissue extracts. Anal Biochem. 2006;356(2):254–64.CrossRefPubMed
16.
Zhu P, Oe T, Blair IA. Determination of cellular redox status by stable isotope dilution liquid chromatography/mass spectrometry analysis of glutathione and glutathione disulfide. Rapid Commun Mass Spectrom. 2008;22(4):432–40.CrossRefPubMed
17.
Harwood DT, Kettle AJ, Brennan S, Winterbourn CC. Simultaneousdetermination of reduced glutathione, glutathione disulfide and glutathionesulphonamide in cells and physiological fluids by isotope dilution liquid chromatography-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci. 2009;877(28):3393–9.CrossRefPubMed
18.
Yan JB, Xu HP, Xiong C, et al. Rapid and reliable detection of glucose-6-phosphate dehydrogenase (G6PD) gene mutations in Han Chinese using high-resolution melting analysis. J Mol Diagn. 2010;12(3):305–11.CrossRefPubMedPubMedCentral
19.
Piraud M, Vianey-Saban C, Petritis K, et al. ESI-MS/MS analysis of underivatised amino acids: a new tool for the diagnosis of inherited disorders of amino acid metabolism. Fragmentation study of 79 molecules of biological interest in positive and negative ionisation mode. Rapid Commun Mass Spectrom. 2003;17(12):1297–311.
20.
Gong ZH, Tian GL, Huang QW, et al. Quantification of total hexose on dry blood spot by tandem mass spectrometry. Clin Biochem. 2012;45(18):1673–7.CrossRefPubMed
21.
Giustarini D, Dalle-Donne I, Lorenzini S, et al. Age-related influence on thiol, disulfide, and protein-mixed disulfide levels in human plasma. J Gerontol A Biol Sci Med Sci. 2006;61(10):1030–8.CrossRefPubMed
22.
Moriarty-Craige SE, Jones DP. Extracellular thiols and thiol/disulfide redox in metabolism. Annu Rev Nutr. 2004;24:481–509.CrossRefPubMed
23.
Johnson JM, Strobel FH, Reed M, et al. A rapid LC-FTMS method for the analysis of cysteine, cystine and cysteine/cystine steady-state redox potential in human plasma. Clin Chim Acta. 2008;396(1–2):43–8.CrossRefPubMedPubMedCentral
24.
Squellerio I, Caruso D, Porro B, et al. Direct glutathione quantification in human blood by LC-MS/MS: comparison with HPLC with electrochemical detection. J Pharm Biomed Anal. 2012;71:111–8.CrossRefPubMed
25.
Steghens JP, Flourié F, Arab K, Collombel C. Fast liquid chromatography-mass spectrometry glutathione measurement in whole blood: micromolar GSSG is a sample preparation artifact. J Chromatogr B Analyt Technol Biomed Life Sci. 2003;798(2):343–9.CrossRefPubMed
26.
Feng CH, Huang HY, Lu CY. Quantitation of the glutathione in human peripheral blood by matrix-assisted laser desorption ionization time-of-flight mass spectrometry coupled with micro-scale derivatization. Anal Chim Acta. 2011;690(2):209–14.CrossRefPubMed
27.
Monostori P, Wittmann G, Karg E, Túri S. Determination of glutathione and glutathione disulfide in biological samples: an in-depth review. J Chromatogr B Analyt Technol Biomed Life Sci. 2009;877(28):3331–46.CrossRefPubMed
28.
Rossi R, Giustarini D, Milzani A, Dalle-Donne I. Membrane skeletal protein S-glutathionylation and hemolysis in human red blood cells. Blood Cells Mol Dis. 2006;37(3):180–7.CrossRefPubMed
Metadata
Title
Reduced glutathione and glutathione disulfide in the blood of glucose-6-phosphate dehydrogenase-deficient newborns
Authors
Zhen-hua Gong
Guo-li Tian
Qi-wei Huang
Yan-min Wang
Hong-ping Xu
Publication date
01-12-2017
Publisher
BioMed Central
Published in
BMC Pediatrics / Issue 1/2017
Electronic ISSN: 1471-2431
DOI
https://doi.org/10.1186/s12887-017-0920-y

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