title: Comprehensive analysis of the lipophilic reactive carbonyls present in biological specimens by LC/ESI-MS/MS.
abstract: A new analytical method has been developed for profiling lipophilic reactive carbonyls (RCs) such as aldehydes and ketones in biological samples using liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) with selected reaction monitoring (SRM). The method consists of several phases, including (1) extraction of lipophilic RCs with a chloroform/methanol mixture; (2) derivatization of the extracted RCs with dansyl hydrazine (DH); and (3) SRM detection of the characteristic product ion of the 5-dimethylaminonaphthalene-1-sulfonyl moiety (m/z 236.1). The analytical results were expressed as RC maps, which allowed for the occurrence and levels of different lipophilic RCs to be visualized. We also developed a highly reproducible and accurate method to extract, purify and derivatize RCs in small volumes of biological specimens. This method was applied to the detection of free RCs in mice plasma samples, and resulted in the detection of more than 400 RCs in samples obtained from C57BL/6J mice. Thirty-four of these RCs were identified by comparison with authentic RCs. This method could be used to investigate the levels of RCs in biological and environmental samples, as well as studying the role of lipid peroxidation in oxidative stress related-disorders and discovering new biomarkers for the early diagnosis of these diseases.
authors.affiliation: Laboratory of Longevity Biochemistry, Graduate School of Integrated Pharmaceutical and Nutritional Sciences, Graduate Program in Food and Nutritional Sciences, University of Shizuoka, Shizuoka 422-8526, Japan. Electronic address: gp1535@u-shizuoka-ken.ac.jp.
authors.forename: Susumu
authors.initials: S
authors.lastname: Tomono
authors.affiliation: Laboratory of Longevity Biochemistry, Graduate School of Integrated Pharmaceutical and Nutritional Sciences, Graduate Program in Food and Nutritional Sciences, University of Shizuoka, Shizuoka 422-8526, Japan.
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Miyoshi
authors.affiliation: Laboratory of Longevity Biochemistry, Graduate School of Integrated Pharmaceutical and Nutritional Sciences, Graduate Program in Food and Nutritional Sciences, University of Shizuoka, Shizuoka 422-8526, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ohshima
mesh_terms: D000447:Aldehydes / Q000032:analysis*; D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D003619:Dansyl Compounds / Q000737:chemistry; D006834:Hydrazines / Q000737:chemistry; D057927:Hydrophobic and Hydrophilic Interactions; D007659:Ketones / Q000032:analysis*; D057230:Limit of Detection; D016014:Linear Models; D015227:Lipid Peroxidation; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000447:Aldehydes; D003619:Dansyl Compounds; D006834:Hydrazines; D007659:Ketones; C027913:dansyl hydrazine
keywords: Aldehyde; Dansyl hydrazine; LC/ESI-MS/MS; Lipid peroxidation; Reactive carbonyl; SRM