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Simultaneous determination of reduced and oxidized glutathione in tissues by a novel liquid chromatography-mass spectrometry method: application in an inhalation study of Cd nanoparticles
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SYSNO ASEP 0434302 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Simultaneous determination of reduced and oxidized glutathione in tissues by a novel liquid chromatography-mass spectrometry method: application in an inhalation study of Cd nanoparticles Author(s) Bláhová, L. (CZ)
Kohoutek, J. (CZ)
Lebedová, J. (CZ)
Bláha, L. (CZ)
Večeřa, Zbyněk (UIACH-O) RID
Buchtová, Marcela (UZFG-Y) RID, ORCID
Míšek, Ivan (UZFG-Y)
Hilscherová, K. (CZ)Number of authors 8 Source Title Analytical and Bioanalytical Chemistry. - : Springer - ISSN 1618-2642
Roč. 406, č. 24 (2014), s. 5867-5876Number of pages 10 s. Publication form Print - P Language eng - English Country DE - Germany Keywords LC MS/MS ; nanoparticle ; cadmium oxide Subject RIV CB - Analytical Chemistry, Separation Subject RIV - cooperation Institute of Animal Physiology and Genetics R&D Projects GAP503/11/2315 GA ČR - Czech Science Foundation (CSF) Institutional support UIACH-O - RVO:68081715 ; UZFG-Y - RVO:67985904 UT WOS 000341834500013 DOI 10.1007/s00216-014-8033-z Annotation The paper presents the development of an advanced extraction and fast analytical LC MS/MS method for simultaneous analyses of reduced and oxidized glutathione (GSH and GSSG, respectively) in different animal tissues. The simultaneous determination of GSH and GSSG is crucial because the amount and ratio of both GSH and GSSG may be altered in response to oxidative stress, an important mechanism of toxicity. The method uses the derivatization of free thiol groups in GSH. Its performance was demonstrated for less explored tissues (lung, brain, and liver) in mouse. The combined extraction and analytical method has very low variability and good reproducibility, maximum coefficients of variance for within-run and between-run analyses under 8 %, and low limits of quantification; for GSH and GSSG, these were 0.2 nM (0.06 ng/mL) and 10 nM (6 ng/mL), respectively. The performance of the method was further demonstrated in a model experiment addressing changes in GSH and GSSG concentrations in lung of mice exposed to CdO nanoparticles during acute 72 h and chronic 13-week exposures. Inhalation exposure led to increased GSH concentrations in lung. GSSG levels were in general not affected; nonsignificant suppression occurred only after the longer 13-week period of exposure. The developed method for the sensitive detection of both GSH and GSSG in very low tissue mass enables these parameters to be studied in cases where only a little sample is available, i.e. in small organisms or in small amounts of tissue. Workplace Institute of Analytical Chemistry Contact Iveta Drobníková, drobnikova@iach.cz, Tel.: 532 290 234 Year of Publishing 2015
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