Previously, it has been shown that
phospholipids,
cholesterol, and
glycolipids could be quantitated using the same high performance thin-layer chromatography (HPTLC) method. Here we examined that method in terms of linearity of standards in the nanogram range, recovery of nonacidic and acidic
lipids after
Sephadex column chromatography, and quantitation of
lipids in mouse synaptic plasma membranes (SPM) where
lipid content is low. Nonacidic and acidic fractions were separated by
Sephadex column chromatography, applied to plates using contact
spotting, chromatographed, visualized with
cupric acetate, and quantitated using in situ densitometry. Recovery of nonacidic and acidic fractions off the columns was determined with radiolabeled
phospholipids. Standards for each
lipid class were linear in the nanogram range. Quantitation of SPM
lipid classes could be made with as little as 1.5 micrograms of total
lipid. Recovery of the nonacidic fraction after
Sephadex column chromatography was approximately 100% whereas the acidic fraction was approximately 91%.
Phospholipids,
cholesterol, and
glycolipids could be determined in nanogram amounts using the same method. This method is an efficient method for examining different
lipid classes and in samples where
lipid content is low.