Measurement of gluconeogenesis using glucose fragments and mass spectrometry after ingestion of deuterium oxide
Measurement of gluconeogenesis using glucose fragments and mass spectrometry after ingestion of deuterium oxide
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DOI：
10.1152/japplphysiol.00752.2007
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发表时间：
2008-04-01
影响因子：
3.3
通讯作者：
Haymond, Morey W.
中科院分区：
文献类型：
--
作者：
Chacko, Shaji K.;Sunehag, Agneta L.;Haymond, Morey W.
报道了一种用气相色谱-质谱法和正化学电离法测定葡萄糖异生作用形成的葡萄糖组分的新方法。受试者摄取了氧化氢后，糖异生作用产生的葡萄糖被用重氢标记。我们对糖异生作用的计算是基于对葡萄糖碳1、3、4、5和6上的氢的平均富集量和人体水中的重氢的富集量的测量。在一名成人志愿者的样本中，用Landau等人的“平均氚富集法”和C-5六亚甲基四胺(HMT)法测定的葡萄糖异生分数分别为48.3+/-0.5%(平均值+/-SD)和C-5-六亚甲基四胺(HMT)法。(Landau BR，Wahren J，Chandramouli V，Schumann WC，Ekberg K，Kalhan SC；J Clin Invest 98：378-385,1996)为46.9+/-5.4%。用新方法对10次重复分析的变异系数为1.0%，而C-5HMT方法的变异系数为11.5%。在接受完全肠外营养的婴儿的样本中，使用新方法的糖异生分数为13.3+/-0.3%，使用C-5 HMT方法的糖异生分数为13.7+/-0.8%。6名成人志愿者连续禁食66h后，用新方法测得的糖异生分数为83.7+/-2.3%，用C-5-HMT法测得的糖异生分数为84.2+/-5.0%。总而言之，平均的氚富集法简单、重复性高、成本效益高。此外，它只需要少量的血液样本。使用额外的示踪剂，也可以在相同的分析过程中测量葡萄糖的出现率。因此，新方法使大量研究人员在所有受试者群体的低糖异生和高糖异生(类似于10到类似于90)的条件下可以进行糖异生测量，并且负担得起。
We report a new method to measure the fraction of glucose derived from gluconeogenesis using gas chromatography-mass spectrometry and positive chemical ionization. After ingestion of deuterium oxide by subjects, glucose derived from gluconeogenesis is labeled with deuterium. Our calculations of gluconeogenesis are based on measurements of the average enrichment of deuterium on carbon 1, 3, 4, 5, and 6 of glucose and the deuterium enrichment in body water. In a sample from an adult volunteer after ingestion of deuterium oxide, fractional gluconeogenesis using the "average deuterium enrichment method" was 48.3 +/- 0.5% (mean +/- SD) and that with the C-5 hexamethylenetetramine (HMT) method by Landau et al. (Landau BR, Wahren J, Chandramouli V, Schumann WC, Ekberg K, Kalhan SC; J Clin Invest 98: 378-385, 1996) was 46.9 +/- 5.4%. The coefficient of variation of 10 replicate analyses using the new method was 1.0% compared with 11.5% for the C-5 HMT method. In samples derived from an infant receiving total parenteral nutrition, fractional gluconeogenesis was 13.3 +/- 0.3% using the new method and 13.7 +/- 0.8% using the C-5 HMT method. Fractional gluconeogenesis measured in six adult volunteers after 66 h of continuous fasting was 83.7 +/- 2.3% using the new method and 84.2 +/- 5.0% using the C-5 HMT method. In conclusion, the average deuterium enrichment method is simple, highly reproducible, and cost effective. Furthermore, it requires only small blood sample volumes. With the use of an additional tracer, glucose rate of appearance can also be measured during the same analysis. Thus the new method makes measurements of gluconeogenesis available and affordable to large numbers of investigators under conditions of low and high fractional gluconeogenesis (similar to 10 to similar to 90) in all subject populations.