Exploratory Biomarkers, Atherosclerosis, Merck Research Laboratories, 126 E. Lincoln Avenue, Rahway, NJ 07065, USA.
Anal Biochem. 2010 Sep 15;404(2):235-7. doi: 10.1016/j.ab.2010.05.010. Epub 2010 May 19.
Measurements of a 2H-labeling of water in biological fluids are required for determining the rates of biochemical flux and for estimating body composition. We have been using the method which relies on the base-catalyzed exchange of hydrogen (deuterium) between water and acetone. 2H-labeling of acetone is then determined using GCMS. Although not noted in the original paper, when chloroform is used to extract the acetone there is slow but substantial back exchange between [2H]acetone and solvent (unpublished observations). We report herein on a refinement of the assay that utilizes headspace analysis, which minimizes the number of transfers and decreases sample preparation time and instrument run time.
测量生物体液中 2H 标记水的含量，对于确定生化通量的速率和估计身体成分是必需的。我们一直在使用依赖于水和丙酮之间的碱基催化交换的氢（氘）的方法。然后使用 GCMS 来确定丙酮的 2H 标记。尽管在原始论文中没有指出，但当使用氯仿提取丙酮时，[2H]丙酮和溶剂之间会发生缓慢但大量的反向交换（未发表的观察结果）。我们在此报告一种改进的测定方法，该方法利用顶空分析，最大限度地减少转移次数，缩短样品制备时间和仪器运行时间。
