Journal of Lipid Research, Vol 29, 1397-1399, Copyright © 1988 by Lipid Research, Inc.
Assay for triacylglycerol lipase by a rapid thin-layer chromatographic technique
MJ Hills and KD Mukherjee
Federal Center for Lipid Research, H. P. Kaufmann-Institute, Munster, Federal Republic of Germany.
A rapid and accurate assay for lipase-catalyzed hydrolysis of radioactively
labeled triacylglycerols has been developed. Aliquots of reaction mixtures
are applied directly, i.e., without extraction of the lipolysis products,
to thin-layer chromatography plates coated with Silica Gel H containing 5%
Na2CO3 (w/w), heated for 10 sec, and developed with diethyl ether-methanol
97:3 (v/v) to a height of 4-5 cm. About 98.5% of the fatty acids are
immobilized as sodium salts at the origin of the chromatogram, whereas
tri-, di-, and monoacylglycerols migrate close to the solvent front.
Adsorbent at the origin and that at the remaining part of the chromatogram
are then assayed for radioactivity without prior staining.
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