Quantification of trans-4-hydroxy-2-nonenal enantiomers and metabolites by LC-ESI-MS/MS.
Lipid peroxidation is a causal factor in multiple diseases including Alzheimer's disease, atherosclerosis, and alcoholic liver disease. One of the most studied products of lipid peroxidation, trans-4-hydroxy-2-nonenal (HNE), has multiple cell signaling and cytotoxic effects. In this work, we developed an LC-MS/MS method for the quantitation of HNE enantiomers, the metabolite trans-4-hydroxy-2-nonenoic acid, and HNE-glutathione adducts in a single chromatographic run. In this method, (R)-HNE and (S)-HNE are derivatized by (S)-carbidopa to form diastereomers that are separated by a reversed-phase column. This method was successfully validated and tested using respiring rat brain mitochondria that enantioselectively metabolize HNE. Metabolic profiles of HNE biotransformation, including the enantiomeric disposition of HNE, will provide useful biomarker data regarding lipid peroxidation in disease states.
Department of Pharmacology, Physiology, and Therapeutics, University of North Dakota, Grand Forks, ND 58203, USA.
Ales
A
Honzatko
Jiri
J
Brichac
Matthew J
MJ
Picklo
D000447:Aldehydes / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D000544:Alzheimer Disease / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001921:Brain / Q000378:metabolism*; D002230:Carbidopa / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005978:Glutathione / Q000378:metabolism*; D015227:Lipid Peroxidation; D008297:Male; D008928:Mitochondria / Q000378:metabolism*; D015394:Molecular Structure; D010084:Oxidation-Reduction; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D013056:Spectrophotometry, Ultraviolet; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry / Q000379:methods
D000447:Aldehydes; D015415:Biomarkers; D005978:Glutathione; C027576:4-hydroxy-2-nonenal; D002230:Carbidopa