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The determination of glutathione-4-hydroxynonenal (GSHNE), E-4-hydroxynonenal (HNE), and E-1-hydroxynon-2-en-4-one (HNO) in mouse liver tissue by LC-ESI-MS

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Abstract

Glutathione (GSH) conjugation of 4-hydroxy-2(E)-nonenal (HNE) is an efficient means of cellular detoxification. HNE is a byproduct of lipid peroxidation which has shown toxicity but also signaling roles. E-1-hydroxynon-2-en-4-one (HNO) is another byproduct of lipid peroxidation which has the same molecular weight as HNE. This study presents the LC-MS detection of GS-HNE, HNE, and HNO in tissue samples without derivatization and with minimal sample preparation. Tissue samples were taken from wild-type mice and knock-out mice, which have been bred without the RLIP76 transfer protein. Extraction procedures were developed to determine GS-HNE and HNE levels in the mouse liver tissue. A gradient elution LC–MS method was developed for GS-HNE analysis using electrospray ionization and selected ion monitoring (SIM). The HNE/HNO method involves isocratic elution due to instability issues. Higher levels of GSHNE, HNE, and HNO were found in the knock-out animals, due to the absence of the RLIP76 transport mechanism.

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, University of Texas at Arlington, Arlington, TX, 76019, USA

    Molly M. Warnke, Eranda Wanigasekara & Daniel W. Armstrong

  2. Department of Molecular Biology & Immunology, University of North Texas Health Science Center, Fort Worth, TX, 76107, USA

    Sharad S. Singhal, Jyotsana Singhal & Sanjay Awasthi

Authors
  1. Molly M. Warnke
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  2. Eranda Wanigasekara
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  3. Sharad S. Singhal
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  4. Jyotsana Singhal
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  5. Sanjay Awasthi
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  6. Daniel W. Armstrong
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Correspondence to Daniel W. Armstrong.

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Warnke, M.M., Wanigasekara, E., Singhal, S.S. et al. The determination of glutathione-4-hydroxynonenal (GSHNE), E-4-hydroxynonenal (HNE), and E-1-hydroxynon-2-en-4-one (HNO) in mouse liver tissue by LC-ESI-MS. Anal Bioanal Chem 392, 1325–1333 (2008). https://doi.org/10.1007/s00216-008-2383-3

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  • DOI: https://doi.org/10.1007/s00216-008-2383-3

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