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Journal of Lipid Research, Vol 24, 1243-1250, Copyright © 1983 by Lipid Research, Inc.
LJ Macala, RK Yu and S Ando
We have devised a high performance thin-layer chromatography (HPTLC)
densitometry method to resolve the major lipid classes of brain tissue. We
used DEAE-Sephadex column chromatography to separate the total lipid into
neutral and acidic lipid fractions. The lipid fractions were then spotted
on separate HPTLC plates and chromatographed in one dimension using two
solvent systems. Quantitation was by in situ densitometry with absolute
amounts of the lipid classes determined from co- chromatographed standards.
An internal standard was also used to improve the precision. The individual
lipid classes of rat whole brain, human brain gray and white matter, rat
and bovine myelin, and bovine oligodendroglia were quantitated. Human brain
phosphatidylethanolamine plasmalogen was also quantitated. Sensitivity was
increased by using the cupric acetate charring reagent, which we found to
be more sensitive than the conventional sulfuric acid-dichromate reagent.
Total lipid (less than 400 micrograms) was quantitated from 5 mg of tissue
wet weight. The limit of detection, on HPTLC, for the individual lipid
classes was below 20 ng.
ARTICLES
Analysis of brain lipids by high performance thin-layer chromatography and densitometry
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