A novel radiochemical method for the microdetermination of total and individual ketone bodies in biological materials

✍ Scribed by Masahiko Ogihara; Yukiko Tokumitsu; Michio Ui

• Publisher: Elsevier Science
• Year: 1978
• Tongue: English
• Weight: 786 KB
• Volume: 85
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(78)90234-8

✦ Synopsis

A novel radiochemical method has been developed for uhramicrodetermination of acetone based on the principle that 1231-labeled iodoform is produced by iodination of acetone with '29CI. ['251]Iodoform is readily counted as a measure of acetone after separation from unreacted iodide ions. Quantitative conversion of 3-hydroxybutyrate to acetoacetate takes place when NAD-dependent oxidation of 3-hydroxybutyrate by 3-hydroxybutyrate dehydrogenase is coupled with NADH-dependent reduction of pyruvate (or 2-oxoglutarate) by lactic dehydrogenase (or glutamic dehydrogenase). Acetoacetate thus formed produces acetone spontaneously when the acidified (deproteinized) reaction mixture is maintained at 50°C for 2 hr. Thus, total and individual ketone bodies in plasma are determined conveniently by combining the radiochemical determination of acetone with these conversion procedures.