Shotgun Lipidomics Analysis of 4-Hydroxyalkenal Species Directly

Apr 15, 2012 - sacrificed by asphyxiation with carbon dioxide or cervical dislocation. Heart, kidney, liver, and skeletal muscle tissues were dissecte...
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Shotgun Lipidomics Analysis of 4-Hydroxyalkenal Species Directly from Lipid Extracts after One-Step in Situ Derivatization Miao Wang, Huafeng Fang, and Xianlin Han* Diabetes and Obesity Research Center, Sanford-Burnham Medical Research Institute, 6400 Sanger Road, Orlando, Florida 32827, United States S Supporting Information *

ABSTRACT: 4-Hydroxyalkenal species are a class of peroxidative products of polyunsaturated fatty acids, which serve as “toxic second messengers” in cellular systems. Investigation of their cellular role is hindered due to the lack of sensitive, reliable, robust method for identification and quantification of these metastable metabolites. Herein, we explored the facile Michael adduct of carnosine with 4hydroxyalkenal species and developed a sensitive, facile, shotgun lipidomics-based method for quantification of these compounds directly from organic solvent lipid extracts of biological samples. In the study, we extensively examined the factors that may affect the accurate quantification of 4hydroxyalkenal species and found that this method possessed high reproducibility (