REVERSED-PHASE RETENTION BEHAVIOR OF FLUORESCENCE-LABELED PHOSPHOLIPIDS IN AMMONIUM ACETATE BUFFERS

Subcomponents of fluorescent derivatives of phosphatidylethanolamine ( PE) and phosphatidylserine (PS) were resolved by reversed-phase high-p erformance liquid chromatography (HPLC) with mobile phases containing acetonitrile, methanol, water, and ammonium acetate. fluorescence labe led phospholipids (PL) include N-(rhodamine B sulfonyl)-PE, N-(7-nitro benz-2-oxa-1,3-diazol-4-yl)-PE, N-(5-fluoresceinthiocarbamoyl) (FL)-PE , N-(1-pyrenesulfonyl) -PE, and N-(5-dimethylaminonaphthalene-1-sulfon yl)-PE. Among the compounds studied, FL-PE exhibited the highest degre e of selectivity for component resolution. The HPLC behavior of the fi ve PE derivatives was examined under variable concentrations of ammoni um acetate. Capacity factors of the PL subcomponents increased with in creasing concentrations of the acetate buffers. Incorporation of triet hylamine into the mobile phase alleviated peak