Schäffer L, Tronier B
Novo Research Institute, Copenhagen, Denmark.
Diabetologia. 1989 Sep;32(9):660-2. doi: 10.1007/BF00274253.
A simple and cheap one-step enzymatic method has been developed for the determination of 1-14C-glucose in plasma. C-1 of glucose is cleaved off as CO2 by treatment with hexokinase, glucose-6-phosphate dehydrogenase, and 6-phosphogluconic dehydrogenase. True 1-14C-glucose activity is then calculated as the difference between total radioactivity and radioactivity remaining after enzyme treatment and evaporation. The reaction is shown to be quantitative and specific, thus eliminating both labelled metabolites and label recycled to other positions in glucose. Two different types of pig experiments show that 1-14C-glucose, when determined by this method, is as irreversible a tracer as the commonly used 3-3H-glucose.
已开发出一种简单且廉价的一步酶法来测定血浆中的1-¹⁴C-葡萄糖。通过己糖激酶、葡萄糖-6-磷酸脱氢酶和6-磷酸葡萄糖酸脱氢酶处理，葡萄糖的C-1以二氧化碳形式被裂解掉。然后将真实的1-¹⁴C-葡萄糖活性计算为总放射性与酶处理和蒸发后剩余放射性之间的差值。该反应具有定量性和特异性，从而消除了标记代谢物以及循环到葡萄糖其他位置的标记。两项不同类型的猪实验表明，用这种方法测定时，1-¹⁴C-葡萄糖与常用的3-³H-葡萄糖一样，是一种不可逆的示踪剂。
