Assay for triacylglycerol lipase by a rapid thin-layer chromatographic technique.
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.
Hills
M J
MJ
Federal Center for Lipid Research, H. P. Kaufmann-Institute, Münster, Federal Republic of Germany.
Mukherjee
K D
KD
D002384:Catalysis; D002855:Chromatography, Thin Layer; D005227:Fatty Acids / Q000302:isolation & purification; D005989:Glycerides / Q000302:isolation & purification; D008049:Lipase / Q000032:analysis*; D014280:Triglycerides / Q000302:isolation & purification
D005227:Fatty Acids; D005989:Glycerides; D014280:Triglycerides; D008049:Lipase