Sl. Abidi et al., REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY OF PHOSPHOLIPIDS WITH FLUORESCENCE DETECTION, Journal of chromatography, 639(2), 1993, pp. 175-184
Reversed-phase high-performance liquid chromatographic (HPLC) behavior
of fluorescent labeled phospholipids (PLs) was studied. Molecular spe
cies of phosphatidylethanolamine (PE) derivatized with fluorescein-, t
hiocarbamoyl-, pyrenesulfonyl-, and dimethylaminonaphthalenesulfonyl (
dansyl)-fluorophores were separated on octadecylsilica with a mobile p
hase of acetonitrile-methanol-water in the presence of tetraalkylammon
ium phosphates (TAAPs). Under similar HPLC conditions, dansylated phos
phatidylserines and PE plasmalogens (ether-linked PLs) were also resol
ved. Incorporation of the fluorescein moiety to the parent PE appeared
to facilitate further resolution of its subcomponents. Subcomponents
of dansylated PE derived from egg phosphatidylcholines were quantifiab
le. Effects of the type and concentration of TAAP on capacity factors
of PL solutes were indicative of an ion-pair separation processes. HPL
C with high-molecular-mass TAAPs favored the separation of the compone
nts that remained unresolved with mobile phases containing low-molecul
ar-mass TAAPs. The HPLC-fluorescence detection method provided a usefu
l approach to quantitative analyses of various PL structures. Composit
ions of PL subcomponents were determined directly from peak areas.