Assay of 3-hydroxybutyrate in the picomolar range.
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.
Ramirez
R
R
Laboratory of Experimental Medicine, Brussels Free University, Belgium.
Sener
A
A
Malaisse
W J
WJ
D020155:3-Hydroxybutyric Acid; D000818:Animals; D002250:Carbon Radioisotopes; D002417:Cattle; D003921:Diabetes Mellitus, Experimental / Q000201:enzymology; D003981:Diazoxide / Q000494:pharmacology; D005969:Glutamate Dehydrogenase / Q000378:metabolism*; D006884:Hydroxybutyrate Dehydrogenase / Q000378:metabolism*; D006885:Hydroxybutyrates / Q000032:analysis*; D007656:Ketoglutaric Acids; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008832:Microchemistry; D011865:Radioisotope Dilution Technique; D051381:Rats; D012242:Rhodobacter sphaeroides / Q000201:enzymology
D002250:Carbon Radioisotopes; D006885:Hydroxybutyrates; D007656:Ketoglutaric Acids; D006884:Hydroxybutyrate Dehydrogenase; D005969:Glutamate Dehydrogenase; D003981:Diazoxide; D020155:3-Hydroxybutyric Acid