J. Lipid Res.
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Journal of Lipid Research, Vol. 13, 284-287, March 1972
Copyright © 1972 by Lipid Research, Inc.

Assay of nanogram levels of triglyceride lipase with a radioactive substrate

Walton H. Marsh and Patrick J. Fitzgerald

Department of Pathology, State University of New York, Downstate Medical Center, Brooklyn, New York 11203

A simple, sensitive procedure for the determination of triglyceride lipase activity has been developed. Nanogram amounts of oleic acid hydrolyzed from commercially available [14C]triolein were readily determined by the counting of the radioactivity of substrate and product after their rapid chromatographic separation on copper hydroxide-impregnated ion-exchange paper. Comparison of the relative amounts of radioactivity of the separated substrate and product gave an estimate of the percentage of hydrolysis of substrate. Comparison of results with a standard of pure lipase enables one to express the amount of hydrolysis in terms of the standard lipase. The results show that measured activity is a linear function of time up to 1 hr of incubation and of amounts of enzyme up to 125 ng. Reproducibility of the test is good.

Supplementary key words pancreatic enzymes • chromatography with copper-impregnated ion-exchange paper

Submitted on July 14, 1971
Accepted on October 4, 1971


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