EVALUATION OF DENSITOMETRIC QUANTITATION OF ETHER PHOSPHOLIPIDS BY HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHY

This paper reports on the use of uni-dimensional high performance thin layer chromatography on silica gel for the separation and detection of ether phospholipids involving in situ reflectance densitometry. Dipping reagents for charring (cupric sulfate-phosphoric acid and manganese chloride-sulfuric acid) or fluorescence enhancement (1-anilino-8-naphthalene sulfonate) were evaluated. Two lipophilic fluorescence reagents (2,5-bis-5′-tert-butylbenzoxazolyl-(2′))thiophene and 4-(N,N-dihexadecyl)amino-7-nitrobenz-2-oxa-1,3-diazole) which move in the mobile phase were tested. The latter reagent gave the more sensitive and stable measurements: nanogram amounts of ether phospholipids could be detected in a reproducible way. Since alkyl lysophospholipids are highly polar molecules, high performance thin layer chromatography is a challenge. Characteristics of sorbents and influence of mobile phases on detection and separation are discussed for the NBD-dihexadecylamine fluorescence detection method. © 1990 Friedr. Vieweg & Sohn Verlagsgesellschaft mbH.