通过液体闪烁计数法测定14C标记的酮体。
Determination of 14C-labelled ketone bodies by liquid-scintillation counting.
作者信息
Mayes P A, Felts J M
出版信息
Biochem J. 1967 Jan;102(1):230-5. doi: 10.1042/bj1020230.
Abstract
- A method of assaying (14)C in ketone bodies present in blood by using liquid-scintillation counting is described. 2. d(-)-beta-Hydroxy[(14)C]butyrate is converted quantitatively into [(14)C]acetoacetate by means of a coupled oxidoreduction reaction involving NAD(+), d(-)-beta-hydroxybutyrate dehydrogenase and malic dehydrogenase in the presence of a high concentration of oxaloacetate. 3. [(14)C]Acetoacetate is decarboxylated to acetone and carbon dioxide which are trapped separately in a double-well flask and counted subsequently. 4. The method permits the determination of (14)C activity in the individual ketone bodies and allows the activity in the carboxyl carbon atoms of acetoacetate or of d(-)-beta-hydroxybutyrate to be assayed separately from the activity in the remainder of the molecule. 5. Recoveries of (14)C-labelled ketone bodies added to blood approach 100% with good reproducibility in replicate analyses.
摘要
- 描述了一种通过液体闪烁计数法测定血液中酮体中¹⁴C的方法。2. 在高浓度草酰乙酸存在下，借助涉及NAD⁺、d(-)-β-羟基丁酸脱氢酶和苹果酸脱氢酶的偶联氧化还原反应，d(-)-β-羟基[¹⁴C]丁酸定量转化为[¹⁴C]乙酰乙酸。3. [¹⁴C]乙酰乙酸脱羧生成丙酮和二氧化碳，它们分别被捕集在双阱烧瓶中，随后进行计数。4. 该方法允许测定各个酮体中的¹⁴C活性，并允许分别测定乙酰乙酸或d(-)-β-羟基丁酸羧基碳原子中的活性与分子其余部分的活性。5. 添加到血液中的¹⁴C标记酮体的回收率接近100%，重复分析具有良好的重现性。
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