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Assay of 3-hydroxybutyrate in the picomolar range
R Ramirez1, A Sener, W J Malaisse
1Laboratory of Experimental Medicine, Brussels Free University, Belgium.
Biochemical Medicine and Metabolic Biology
|April 1, 1991
Abstract:
A radioisotopic procedure for the assay of 3-hydroxybutyrate is presented. It is based on the measurement of NADH, generated in the 3-hydroxybutyrate dehydrogenase reaction, through the conversion of 2-[U-14C]ketoglutarate to 14C-labeled L-glutamate in the presence of beef liver glutamate dehydrogenase. The assay is linear in the range of 2.5 to 20.0 pmole/sample and about 100-times more sensitive than previous methods. The procedure proved useful for the measurement of 3-hydroxybutyrate in liver samples not exceeding 25 micrograms wet weight.