Department of Pediatrics, USDA/Agricultural Research Service Children's Nutrition Research Center, Baylor College of Medicine, Houston, TX, USA.
J Nutr. 2012 Nov;142(11):2057-62. doi: 10.3945/jn.112.167957. Epub 2012 Sep 26.
Stable hydrogen isotope methodology is used in nutrition studies to measure growth, breast milk intake, and energy requirement. Isotope ratio MS is the best instrumentation to measure the stable hydrogen isotope ratios in physiological fluids. Conventional methods to convert physiological fluids to hydrogen gas (H(2)) for mass spectrometric analysis are labor intensive, require special reagent, and involve memory effect and potential isotope fractionation. The objective of this study was to determine the accuracy and precision of a platinum catalyzed H(2)-water equilibration method for stable hydrogen isotope ratio measurements. Time to reach isotopic equilibrium, day-to-day and week-to-week reproducibility, accuracy, and precision of stable hydrogen isotope ratio measurements by the H(2)-water equilibration method were assessed using a Thermo DELTA V Advantage continuous-flow isotope ratio mass spectrometer. It took 3 h to reach isotopic equilibrium. The day-to-day and week-to-week measurements on water and urine samples with natural abundance and enriched levels of deuterium were highly reproducible. The method was accurate to within 2.8 (o)/oo and reproducible to within 4.0 (o)/oo based on analysis of international references. All the outcome variables, whether in urine samples collected in 10 doubly labeled water studies or plasma samples collected in 26 body water studies, did not differ from those obtained using the reference zinc reduction method. The method produced highly accurate estimation on ad libitum energy intakes, body composition, and water turnover rates. The method greatly reduces the analytical cost and could easily be adopted by laboratories equipped with a continuous-flow isotope ratio mass spectrometer.
稳定氢同位素方法学被用于营养研究中,以测量生长、母乳摄入量和能量需求。同位素比质谱是测量生理流体中稳定氢同位素比值的最佳仪器。将生理流体转化为氢气(H2)进行质谱分析的传统方法劳动强度大,需要特殊试剂,并且涉及记忆效应和潜在的同位素分馏。本研究的目的是确定铂催化 H2-水平衡方法用于稳定氢同位素比测量的准确性和精密度。使用 Thermo DELTA V Advantage 连续流动同位素比质谱仪评估了达到同位素平衡的时间、日常和每周重复性、稳定氢同位素比测量的准确性和精密度。达到同位素平衡需要 3 小时。对具有天然丰度和氘富集水平的水和尿液样品进行的日常和每周测量具有高度重现性。该方法在分析国际参考物质时的准确性在 2.8(o)/oo 以内,重现性在 4.0(o)/oo 以内。无论是在 10 项双标记水研究中收集的尿液样本还是在 26 项体液研究中收集的血浆样本中的所有结果变量,与使用参考锌还原法获得的结果无差异。该方法对随意能量摄入、身体成分和水周转率的估计非常准确。该方法大大降低了分析成本,并且配备连续流动同位素比质谱仪的实验室很容易采用。