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Clinical Chemistry, Vol 19, 476-482, Copyright © 1973 by the American Association for Clinical Chemistry
1 From the Biochemical Research Laboratories, Calbiochem,
10933 North Torrey Pines Road, Calif. 92037.
We describe a novel method for determining serum triglycerides, in which an enzymatic hydrolysis replaces the more commonly used saponification procedure. Under the conditions of the assay, the enzymatic hydrolysis can be completed in less than 10 min by the combined action of a microbial lipase and a protease. We have been able to demonstrate complete hydrolysis of triglycerides by thin-layer chromatography of the reaction products, by recovery of glycerol from sera of known triglycerides content, and by comparison of triglyceride assays on a number of sera assayed by our method vs. the AutoAnalyzer procedure. The hydrolysis is directly coupled to the enzymatic determination of glycerol, and is followed through absorbance changes at 340 nm. The assay is simple, rapid, and requires only 50 µl or less of sample. Because the enzymes used do not release glycerol from other compounds in serum, the hydrolysis can be considered specific for triglycerides.
-chymotrysin determination of glycerol by coupled enzymatic
reaction thin-layer chromatography AutoAnalyzer
Submitted on May 23, 1972
Accepted on February 13, 1973
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