High-performance liquid chromatography and laser desorption/ionization mass spectrometry of retinyl esters.
Laser desorption/ionization mass spectrometry (LDI-MS) at 337 nm laser wavelength was used to analyze retinol and several long-chain fatty acid esters of retinol. Employing this ionization technique helped to overcome the inherent problems resulting from thermal instability of retinyl esters which render this group of compounds rather difficult for standard ionization techniques. Mass spectra were recorded with a linear time-of-flight instrument in positive ion mode. Under these conditions, retinyl esters formed radical molecular ions (M.+) and in addition fragmented by elimination of the fatty acyl chain to uniformly form a peak at m/z = 269 u. The elimination of carbon dioxide was also observed in the spectra. The method is suitable to identify specific retinyl esters in complex mixtures of these compounds. A gradient reversed-phase high-performance liquid chromatography (HPLC) is described for the separation of retinol and 15 related fatty acid esters within 28 min. HPLC was applied to separate retinyl esters from rat liver extracts. The LDI mass spectrum of the collected HPLC fraction of rat liver extract showed the molecular parent ions of retinyl myristate, pentadecanoate, palmitoleate, palmitate, heptadecanoate, linoleate, oleate, stearate, and 3,4-didehydroretinyl palmitate. LDI-MS was found to be more appropriate than matrix-assisted laser desorption/ionization for the described analytical task.
Wingerath
T
T
Institut für Physiologische Chemie I, Heinrich-Heine-Universität, Düsseldorf, Germany.
Kirsch
D
D
Spengler
B
B
Kaufmann
R
R
Stahl
W
W
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004952:Esters / Q000737:chemistry; D008099:Liver / Q000737:chemistry; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D012176:Retinoids / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D004952:Esters; D012176:Retinoids