Structure elucidation of retinoids in biological samples using postsource decay laser desorption/ionization mass spectrometry after high-performance liquid chromatography separation.
Retinoids [retinol (vitamin A) and its metabolites] function in the visual cycle, embryonic development, cellular differentiation, and tissue homeostasis. Notwithstanding pivotal roles of retinoids in mammals, the limited number of commercially available retinoid standards is a major roadblock to identifying and studying retinoids in biological samples. Therefore, a need exists for improved methods to identify retinoid metabolites. We analyzed polar and nonpolar retinoids, including retinoic acid, retinol, retinyl acetate, and other retinyl esters, using postsource decay laser desorption/ionization mass spectrometry (PSD-LDI MS). PSD analysis was employed to examine the PSD fragmentation patterns of retinoids, as these patterns can be used for the characterization of retinoids from biological samples without the need for matching retention time with a commercially available or synthetic retinoid. Mechanisms for the formation of these PSD fragment ions are proposed. The feasibility of employing PSD after HPLC separation was demonstrated by characterizing the endogenous retinoids in canine kidney epithelial cell extracts and in mouse lung. We show that the PSD-LDI MS approach described here can facilitate the identification and characterization of retinoids from mammalian cells and tissues.
Suh
Moo-Jin
MJ
Department of Pharmacology, Weill Medical College of Cornell University, 1300 York Avenue, New York, New York 10021, USA.
Tang
Xiao-Han
XH
Gudas
Lorraine J
LJ
D000818:Animals; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D004285:Dogs; D008297:Male; D051379:Mice; D012176:Retinoids / Q000737:chemistry* / Q000302:isolation & purification; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D012176:Retinoids