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.
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.
Warnke
Molly M
MM
Department of Chemistry and Biochemistry, University of Texas at Arlington, Arlington, TX 76019, USA.
Wanigasekara
Eranda
E
Singhal
Sharad S
SS
Singhal
Jyotsana
J
Awasthi
Sanjay
S
Armstrong
Daniel W
DW
D000447:Aldehydes / Q000032:analysis*; D000818:Animals; D002853:Chromatography, Liquid; D020690:GTPase-Activating Proteins / Q000235:genetics / Q000378:metabolism; D005978:Glutathione / Q000032:analysis*; D015227:Lipid Peroxidation; D008099:Liver / Q000737:chemistry*; D051379:Mice; D018345:Mice, Knockout; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D000447:Aldehydes; D020690:GTPase-Activating Proteins; C490620:Ralbp1 protein, mouse; D005978:Glutathione; C027576:4-hydroxy-2-nonenal