Barrie A, Coward W A
Biomed Mass Spectrom. 1985 Sep;12(9):535-41. doi: 10.1002/bms.1200120916.
The doubly labelled water method involves the administration of water enriched in 2H and 18O followed by determination of the turnover rates of these isotopes. Since 18O is eliminated from the body as both CO2 and water, while 2H leaves only as water, the difference between the two turnover rates provides a measure of CO2 production and hence energy expenditure. Isotopic analysis by conventional stable isotope ratio analysis (SIRA) is labour intensive and time consuming, as it requires off-line conversion of water samples to gases (H2 and CO2) followed by sequential analysis for each of the two isotopes using the mass spectrometer. Lack of suitable automated instrumentation with the ability to process large numbers of samples has prevented routine application of the method. We describe here an automated technique in which body water samples (urine, saliva, breath water or milk) are analysed simultaneously for 2H and 18O. The single bench system comprises two mass spectrometer analysers, one for measuring 2H from H2 gas, the other for measuring 18O from the water vapour (masses 18, 20). Both analysers share a common heated inlet system into which microlitre quantities of the body fluids are injected from an autosampler (102 samples). The water vapour flows both directly to one analyser for 18O measurement and into a uranium reduction furnace for conversion to H2, prior to 2H measurement by the second analyser. Both analysers also share vacuum and electronic components, enabling savings in both space and cost. In this paper we present results illustrating performance characteristics and procedures for routine application to human subjects.(ABSTRACT TRUNCATED AT 250 WORDS)
双标记水法包括给予富含氘(2H)和氧-18(18O)的水,然后测定这些同位素的转换率。由于18O以二氧化碳和水的形式从体内排出,而2H仅以水的形式排出,两种转换率之间的差异提供了二氧化碳产生量的度量,从而也反映了能量消耗。通过传统的稳定同位素比率分析(SIRA)进行同位素分析既费力又耗时,因为它需要将水样离线转化为气体(氢气和二氧化碳),然后使用质谱仪对两种同位素分别进行顺序分析。缺乏能够处理大量样品的合适自动化仪器阻碍了该方法的常规应用。我们在此描述一种自动化技术,可同时对人体水样(尿液、唾液、呼出水或乳汁)中的2H和18O进行分析。单台系统包括两个质谱仪分析仪,一个用于测量氢气中的2H,另一个用于测量水蒸气中的18O(质量数18、20)。两个分析仪共用一个加热进样系统,从自动进样器(102个样品)中注入微升量的体液。水蒸气既直接流向一个分析仪进行18O测量,又流入铀还原炉转化为氢气,然后由第二个分析仪进行2H测量。两个分析仪还共用真空和电子组件,从而节省了空间和成本。在本文中,我们展示了该技术的性能特征以及对人类受试者进行常规应用的程序。(摘要截选至250字)