Simultaneous quantification of retinol, retinal, and retinoic acid isomers by high-performance liquid chromatography with a simple gradiation.
A new method of high-performance liquid chromatography (HPLC) analysis to quantify isomers of retinol, retinal and retinoic acid simultaneously was established. The HPLC system consisted of a silica gel absorption column and a linear gradient with two kinds of solvents containing n-Hexane, 2-propanol, and glacial acetic acid in different ratios. It separated six retinoic acid isomers (13-cis, 9-cis, all-trans, all-trans-4-oxo, 9-cis-4-oxo, 13-cis-4-oxo), three retinal isomers (13-cis-, 9-cis-, and all-trans) and two retinol isomers (13-cis- and all-trans). Human serum samples were subjected to this HPLC analysis and at least, all-trans retinol, 13-cis retinol, and all-trans retinoic acid were detectable. This HPLC system is useful for evaluating retinoic acid formation from retinol via a two-step oxidation pathway. Moreover, it could be applied to monitoring the concentrations of various retinoids, including all-trans retinoic acid in human sera.
Miyagi
M
M
Department of Internal Medicine, School of Medicine, Keio University, Tokyo, Japan.
Yokoyama
H
H
Shiraishi
H
H
Matsumoto
M
M
Ishii
H
H
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D007536:Isomerism; D012172:Retinaldehyde / Q000032:analysis*; D013056:Spectrophotometry, Ultraviolet; D014212:Tretinoin / Q000032:analysis*; D014801:Vitamin A / Q000032:analysis*
D014801:Vitamin A; D014212:Tretinoin; D012172:Retinaldehyde