Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Body composition, energy expenditure and physical activity

Maximizing precision and accuracy of the doubly labeled water method via optimal sampling protocol, calculation choices, and incorporation of 17O measurements

European Journal of Clinical Nutrition volume 74, pages 454–464 (2020)Cite this article

Subjects

Abstract

Background/Objectives

The doubly labeled water (DLW) method is the gold standard methodology for determination of free-living, total daily energy expenditure (TEE). However, there is no single accepted approach for either the sampling protocols (daily vs. two-point, in which samples are collected after dosing and at the end of the measurement period) or the calculations used in the determination of the rate of carbon dioxide production (rCO2) and TEE. Moreover, fluctuations in natural background abundances introduce error in the calculation of rCO2 and TEE. The advent of new technologies makes feasible the possibility of including additional isotope measures (17O) to account for background variation, which may improve accuracy.

Subjects/Methods

Sixteen subjects were studied for 7 consecutive days in a whole-room indirect calorimeter (IC) with concurrent measurement of TEE by DLW. Daily urine samples were obtained and isotope ratios were determined using off-axis integrated cavity output spectroscopy (OA-ICOS).

Results

We determined the best combination of approaches for estimating dilution spaces and elimination rates and calculated average daily volume of carbon dioxide production (VCO2) using six different published equations. Using this best combination, multi-point fitting of isotope elimination rates using the daily urine samples substantially improved the average precision (4.5% vs. 6.0%) and accuracy (−0.5% vs. −3.0%) compared with the two-point method. This improvement may partly reflect the less variable day-to-day chamber measurements of energy expenditure. Utilizing 17O measurements to correct for errors due to background isotope fluctuations provided additional but minor improvements in precision (4.2% vs. 4.5%) and accuracy (0.2% vs. 0.5%).

Conclusions

This work shows that optimizing sampling and calculation protocols can improve the accuracy and precision of DLW measurements.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Lifson N, McClintock R. Theory of the use of the turnover rates of body water for measuring energy expenditure. J Theor Biol 1966;12:46–74.

    Article  CAS  Google Scholar 

  2. Speakman JR. Doubly labelled water: theory and practice. London: Chapman and Hall; 1997.

    Google Scholar 

  3. Weir JB. New methods for calculating metabolic rate with special reference to protein metabolism. J Physiol 1949;109:1–9.

    Article  Google Scholar 

  4. Schoeller DA, Ravussin E, Schutz Y, Acheson KJ, Baertschi P, Jequier E. Energy expenditure by doubly labeled water: validation in humans and proposed calculation. Am J Physiol 1986;250(5 Pt 2):R823–30.

    CAS  PubMed  Google Scholar 

  5. Speakman JR, Nair KS, Goran MI. Revised equations for calculating CO2 production from doubly labeled water in humans. Am J Physiol 1993;264(6 Pt 1):E912–7.

    CAS  PubMed  Google Scholar 

  6. Pontzer H. Method and rationale for recalculating dilution spaces to a single, common time point in doubly labeled water studies. Eur J Clin Nutr 2018;72:1620–4.

    Article  CAS  Google Scholar 

  7. Roberts SB, Dietz W, Sharp T, Dallal GE, Hill JO. Multiple laboratory comparison of the doubly labeled water technique. Obes Res 1995;3(Suppl 1):3–13.

    Article  Google Scholar 

  8. Klein PD, James WP, Wong WW, Irving CS, Murgatroyd PR, Cabrera M, et al. Calorimetric validation of the doubly-labelled water method for determination of energy expenditure in man. Hum Nutr Clin Nutr 1984;38:95–106.

    CAS  PubMed  Google Scholar 

  9. Speakman JR, Racey PA. Measurement of CO2 production by the doubly labeled water technique. J Appl Physiol 1986;61:1200–2.

    Article  CAS  Google Scholar 

  10. Speakman JR, Racey PA. Doubly labelled water: errors in the evaluation of oxygen isotope turnover due to temporal variation in CO2 production do not always covary with dilution space estimate. J Theor Biol 1989;141:547–56.

    Article  Google Scholar 

  11. Speakman JR, Racey PA. The equilibrium concentration of oxygen-18 in body water: Implications for the accuracy of the doubly-labelled water technique and a potential new method of measuring RQ in free-living animals. J Theor Biol 1987;127:79–95.

    Article  Google Scholar 

  12. Horvitz MA, Schoeller DA. Natural abundance deuterium and 18-oxygen effects on the precision of the doubly labeled water method. Am J Physiol Endocrinol Metab 2001;280:E965–72.

    Article  CAS  Google Scholar 

  13. Berman ES, Melanson EL, Swibas T, Snaith SP, Speakman JR. Inter- and intraindividual correlations of background abundances of (2)H, (18)O and (17)O in human urine and implications for DLW measurements. Eur J Clin Nutr 2015;69:1091–8.

    Article  CAS  Google Scholar 

  14. Speakman JR. The role of technology in the past and future development of the doubly labelled water method. Isot Environ Health Stud 2005;41:335–43.

    Article  CAS  Google Scholar 

  15. Speakman JR, Krol E. Comparison of different approaches for the calculation of energy expenditure using doubly labeled water in a small mammal. Physiol Biochem Zool 2005;78:650–67.

    Article  Google Scholar 

  16. Berman ES, Fortson SL, Snaith SP, Gupta M, Baer DS, Chery I, et al. Direct analysis of delta2H and delta18O in natural and enriched human urine using laser-based, off-axis integrated cavity output spectroscopy. Anal Chem 2012;84:9768–73.

    Article  CAS  Google Scholar 

  17. Berman ES, Levin NE, Landais A, Li S, Owano T. Measurement of delta18O, delta17O, and 17O-excess in water by off-axis integrated cavity output spectroscopy and isotope ratio mass spectrometry. Anal Chem 2013;85:10392–8.

  18. Melanson EL, Swibas T, Kohrt WM, Catenacci VA, Creasy SA, Plasqui G, et al. Validation of the doubly labeled water method using off-axis integrated cavity output spectroscopy and isotope ratio mass spectrometry. Am J Physiol Endocrinol Metab 2018;314:E124–30.

  19. Coward WA, Prentice AM. Isotope method for the measurement of carbon dioxide production rate in man. Am J Clin Nutr 1985;41:659–63.

    Article  CAS  Google Scholar 

  20. Schoeller DA. Measurement of energy expenditure in free-living humans by using doubly labeled water. J Nutr 1988;118:1278–89.

    Article  CAS  Google Scholar 

  21. Racette SB, Schoeller DA, Luke AH, Shay K, Hnilicka J, Kushner RF. Relative dilution spaces of 2H- and 18O-labeled water in humans. Am J Physiol 1994;267(4 Pt 1):E585–90.

    CAS  PubMed  Google Scholar 

  22. Sagayama H, Yamada Y, Racine NM, Shriver TC, Schoeller DA, Group DLWS. Dilution space ratio of 2H and 18O of doubly labeled water method in humans. J Appl Physiol 2016;120:1349–54.

    Article  CAS  Google Scholar 

  23. Speakman JR. Calculation of CO2 production in doubly-labelled water studies. J Theor Biol 1987;126:101–4.

    Article  CAS  Google Scholar 

  24. Speakman JR. How should we caclulate CO2 production in doubly labeled water studies of animals. Func Ecol 1993;7:746–50.

    Google Scholar 

  25. Krol E, Speakman JR. Isotope dilution spaces of mice injected simultaneously with deuterium, tritium and oxygen-18. J Exp Biol 1999;202(Pt 20):2839–49.

    CAS  PubMed  Google Scholar 

  26. Ritz P, Johnson PG, Coward WA. Measurements of 2H and 18O in body water: analytical considerations and physiological implications. Br J Nutr 1994;72:3–12.

    Article  CAS  Google Scholar 

Download references

Funding

This work was supported by an NIH Small Business Innovation (SBIR) Research Grant (R44 DK093362), as well as support from the Colorado Nutrition and Obesity Research Center (P30 DK048520) and the Colorado Clinical and Translational Science Institute (UL1 RR025780). ELM is also supported by resources from the Geriatric Research, Education, and Clinical Center at the Denver VA Medical Center. JRS was supported by a 1000 Talents award from the Chinese government and a Wolfson merit award from the UK royal society.

Author information

Authors and Affiliations

  1. Kairos Aerospace, Mountain View, CA, USA

    Elena S. F. Berman

  2. Division of Geriatric Medicine, Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, CO, USA

    Tracy Swibas, Wendy M. Kohrt & Edward L. Melanson

  3. Geriatric Research, Education, and Clinical Center, VA Eastern Colorado Health Care System, Denver, CO, USA

    Wendy M. Kohrt & Edward L. Melanson

  4. Division of Endocrinology, Metabolism, and Diabetes, Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, CO, USA

    Vicki A. Catenacci, Seth A. Creasy & Edward L. Melanson

  5. Institute of Biological and Environmental Sciences, Aberdeen University, Aberdeen, UK

    John R. Speakman

  6. State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China

    John R. Speakman

Authors
  1. Elena S. F. Berman
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tracy Swibas
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wendy M. Kohrt
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Vicki A. Catenacci
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Seth A. Creasy
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Edward L. Melanson
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. John R. Speakman
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to John R. Speakman.

Ethics declarations

Conflict of interest

At the time the research was conducted, ESFB was employed by ABB/Los Gatos Research, the company that manufactures the OA-ICOS analyzer used for the analyses in this paper. The others declare that they have no conflict of interest.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Berman, E.S.F., Swibas, T., Kohrt, W.M. et al. Maximizing precision and accuracy of the doubly labeled water method via optimal sampling protocol, calculation choices, and incorporation of 17O measurements. Eur J Clin Nutr 74, 454–464 (2020). https://doi.org/10.1038/s41430-019-0492-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41430-019-0492-z

This article is cited by

Search

Advanced search

Quick links