Skip to main content

Main menu

  • Home
  • Current issue
  • ERJ Early View
  • Past issues
  • ERS Guidelines
  • Authors/reviewers
    • Instructions for authors
    • Submit a manuscript
    • Open access
    • Peer reviewer login
    • WoS Reviewer Recognition Service
  • Alerts
  • Subscriptions
  • ERS Publications
    • European Respiratory Journal
    • ERJ Open Research
    • European Respiratory Review
    • Breathe
    • ERS Books
    • ERS publications home

User menu

  • Log in
  • Subscribe
  • Contact Us
  • My Cart

Search

  • Advanced search
  • ERS Publications
    • European Respiratory Journal
    • ERJ Open Research
    • European Respiratory Review
    • Breathe
    • ERS Books
    • ERS publications home

Login

European Respiratory Society

Advanced Search

  • Home
  • Current issue
  • ERJ Early View
  • Past issues
  • ERS Guidelines
  • Authors/reviewers
    • Instructions for authors
    • Submit a manuscript
    • Open access
    • Peer reviewer login
    • WoS Reviewer Recognition Service
  • Alerts
  • Subscriptions

Effect of oxygen on breath markers of oxidative stress

M. Phillips, R.N. Cataneo, J. Greenberg, R. Grodman, R. Gunawardena, A. Naidu
European Respiratory Journal 2003 21: 48-51; DOI: 10.1183/09031936.02.00053402
M. Phillips
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
R.N. Cataneo
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
J. Greenberg
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
R. Grodman
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
R. Gunawardena
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
A. Naidu
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Abstract

Supplemental oxygen is often administered to induce hyperoxia in nonhypoxic patients for indications such as chest pain, despite lack of evidence of clinical benefit. Induced hyperoxia is potentially toxic, since it may increase oxidative stress and peroxidative damage to deoxyribonucleic acid, lipids and proteins.

The aim of this study was to establish whether supplemental oxygen induces oxidative stress in nonhypoxic subjects.

Breath markers of oxidative stress were measured in 31 healthy subjects before and after breathing 28% oxygen at 2.0 L·min−1 via nasal prongs for 30 min while resting. The criterion standard of oxidative stress was the breath methylated alkane contour (BMAC), a three-dimensional plot of the alveolar gradients of C4–C20 alkanes and monomethylated alkanes produced by lipid peroxidation. Volatile organic compounds (VOCs) in breath were assayed by gas chromatography and mass spectroscopy, and the BMACs before and after oxygenation were compared.

Following oxygenation, there was a significant increase in mean volume under the curve of the BMAC and in alveolar gradients of three VOCs: 3‐methyltridecane, 3‐methylundecane and 5-methylnonane.

Breath markers of oxidative stress were significantly increased in normal volunteers breathing supplemental oxygen for 30 min.

  • breath
  • oxidative stress
  • oxygen breathing
  • volatile organic compounds

Oxygen is a toxic drug. Toxic effects of oxygen were reported in laboratory animals in 1878 but were not recognised in humans until the late 1940s, when an epidemic of blindness in premature newborns was traced to retrolental fibroplasia induced by supplemental oxygen therapy 1. Oxygen at high partial pressures is now recognised as toxic to the respiratory, cardiovascular, nervous and gastrointestinal systems 2. The source of oxygen toxicity arises from the mitochondria, where oxygen is the final acceptor of electrons in oxidative metabolism. Oxygen is reduced to water, as well as a number of reactive oxygen species (ROS), including superoxide radical, hydrogen peroxide and hydroxyl radical. These ROS leak out of the mitochondria into the cytoplasm where they cause oxidative damage to a wide variety of biologically important molecules, including deoxyribonucleic acid (DNA), proteins and polyunsaturated fatty acids (PUFAs) in cell membranes 3, 4 The collective oxidative effect of ROS on biological molecules is termed oxidative stress. It has been implicated as a mechanism of aging 1 and carcinogenesis 5, and is increased in several disorders including rheumatoid arthritis 6, ischaemic heart disease 7 and bronchial asthma 8.

It is a common clinical practice to administer supplemental oxygen in order to induce hyperoxia in nonhypoxic patients with chest pain and suspected acute myocardial infarction. There is little evidence that this is beneficial and it may potentially be harmful by inducing an increase in oxidative stress. However, the effects of oxygen on markers of oxidative stress have not been totally elucidated because previous studies have yielded conflicting results 9–12. The current authors have recently reported a sensitive new index of oxidative stress, the breath methylated alkane contour (BMAC), comprising a three-dimensional surface plot of the abundance in breath of C4–C20 alkanes and their monomethylated derivatives 13. In this study, the effects of supplemental oxygen in a group of healthy volunteers, as shown by induced changes in their BMACs, are reported.

Materials and methods

Human subjects and study design

Healthy volunteers were recruited from the medical staff of Saint Vincents Catholic Medical Centers of New York (Staten Island Region, NY, USA). The 31 subjects comprised 20 males and 11 females; their mean age was 28.4 yrs (sd 5.0). After an overnight fast, each subject breathed 28% oxygen at 2.0 L·min−1 via nasal prongs for 30 min while resting comfortably. Breath samples were collected before and immediately after breathing oxygen. The research was approved by the institutional review board of Saint Vincents Catholic Medical Centers of New York and all subjects gave their signed informed consent to participate.

Breath volatile organic compounds collection and assay

The method has been described previously 14. A portable breath collection apparatus was employed to capture the volatile organic compounds (VOCs) in 1.0 L breath and in 1.0 L room air on to separate sorbent traps. Subjects wore a noseclip while breathing in and out of the disposable mouthpiece of the apparatus for 2.0 min. VOCs in the sorbent traps were analysed by automated thermal desorption, gas chromatography and mass spectroscopy.

Derivation of breath methylated alkane contour

Three-dimensional surface plots of C4–C20 n‐alkanes and their monomethylates were obtained using a previously described method 13. In summary, data were obtained from VOC peaks observed on breath chromatograms: Vb=area under the curve (AUC) of alkane or methylated alkane; Ib=AUC of internal standard used to calibrate the instrument (0.25 mL, 2 parts per million 1‐bromo‐4‐fluoro-benzene (Supelco, Bellefonte, PA, USA)). Va and Ia were corresponding AUCs derived from chromatograms of room air. The alveolar gradient 15, 16 of each VOC was then determined as follows:Embedded ImageMean alveolar gradients of these VOCs were computed for all subjects before and after breathing oxygen and displayed in surface plots showing the carbon chain length on the x‐axis, the methylation site on the z‐axis, and the mean alveolar gradient on the y‐axis.

Analysis of data

Paired t‐tests were employed to compare the pre- and post-oxygenation alveolar gradients of C4–C20 n‐alkanes and their monomethylates, as well as the volume under the curve of the BMACs.

Results

All subjects completed the study without adverse effects. The mean pre- and post-oxygenation BMACs are shown in figure 1⇓. There was a global increase in the alveolar gradients of C4–C20 n‐alkanes and their monomethylated derivatives, as shown by a significant increase in the mean volume under the curve of the BMAC (fig. 2⇓). The alveolar gradients of three BMAC VOCs, 3‐methyltridecane, 3‐methylundecane and 5‐methylnonane, were significantly (p<0.05) increased after oxygenation.

Fig. 1.—
  • Download figure
  • Open in new tab
  • Download powerpoint
Fig. 1.—

Breath methylated alkane contours (BMACs) before and after oxygenation. The BMAC is a three-dimensional surface plot of the results of the breath test for volatile organic compounds (VOC). The mean alveolar gradient (concentration in breath‐concentration in room air) is shown on the vertical axis for C4–C20 alkanes and their monomethylated derivatives. The horizontal axes identify the specific VOC (e.g. the combination of carbon chain length=4 and methylation site=2 corresponds to 2‐methylbutane). It is apparent that the mean alveolar gradients of several VOCs were increased by oxygenation. There was a significant global increase in volume under the curve (see fig. 2⇓).

Fig. 2.—
  • Download figure
  • Open in new tab
  • Download powerpoint
Fig. 2.—

Mean volume under the curve (VUC) of the breath methylated alkane contour (BMAC) before and after oxygenation. This diagram displays the mean VUC of the BMACs in figure 1⇑ (error bar=sem). The change demonstrates a significant global increase in the volatile markers of oxidative stress that comprise the BMAC.

Discussion

Normal healthy volunteers significantly increased their output of a broad spectrum of breath markers of oxidative stress after breathing 28% oxygen for 30 min. These findings are noteworthy, first, because previous studies of hyperoxia have yielded contradictory results, and secondly, because of the clinical implications of monitoring oxygen therapy.

The BMAC comprises a set of 107 different C4–C20 alkanes and monomethylated alkanes, each of which is an independent marker of oxidative stress. These markers were evaluated before and after oxygen breathing in two different ways: first, by comparing changes in the abundance of individual VOCs, and second, by comparing their integrated total abundance as the mean volume under the curve of the BMAC. Three of these individual markers were significantly increased by oxygen breathing, and there was also a significant increase in their integrated total abundance, the mean volume under the curve of the BMAC.

Many different markers of oxidative stress have been proposed, including malonaldehyde and conjugated dienes in the blood, as well as alkanes and hydrogen peroxide in the breath 3, 4. Despite their chemical diversity, all of these markers of oxidative stress share one common feature: all are downstream metabolites of parent molecules that have been oxidised by ROS. However, no comprehensive study has yet compared the sensitivity and specificity of these different markers or estimated their relative value as clinical research tools. Consequently, it would be desirable to validate the current findings in future studies by comparing the effects of oxygen breathing on other markers of oxidative stress.

Previous studies of the effects of hyperoxia on other markers of oxidative stress have yielded inconsistent findings. Hyperoxia in normal humans breathing 100% oxygen has been reported to increase breath levels of pentane as well as serum malondialdehyde 9, 10 but induced hyperoxia in newborn lambs elicited no significant changes in either breath ethane or pentane 17. In preterm infants ventilated for respiratory distress syndrome, Drury et al. 18 found no correlation between fractional inspired oxygen concentration and exhaled pentane, but Nycyk et al. 19 found an association between high-peak pentane exhalation and low gestational age, mortality, intraventricular haemorrhage and retinopathy of prematurity.

The inconsistency of these findings has led to controversy as to whether or not oxygen breathing is the real cause of increased oxidative stress. However, these inconsistencies may have resulted, at least in part, from technically inadequate measurements. While breath testing may appear intuitive and simple, its apparent simplicity conceals a number of formidable obstacles that can result in technical failure. Technical failures in breath testing for markers of oxidative stress are of three main types. 1) Inadequate breath collection technique and failure to correct for VOCs present in background air. Cailleux and Allain 20 observed that pentane is a normal constituent of room air. Hence, any breath test that is not corrected for VOCs inspired from room air will generate erroneous results. 2) Inadequate analytic technique. Kohlmuller and Kochen 21 found that commonly used chromatography columns failed to separate n‐pentane and isoprene, and they concluded that reports of breath pentane in the literature had actually measured isoprene or a mixture of pentane and isoprene. 3) Failure to recognise the inherent limitations of employing a single-breath marker of oxidative stress. Ethane and pentane in breath may be measured (or mismeasured) quickly and easily. This has resulted in a widespread misconception that ethane and pentane are the sole markers of oxidative stress in breath, while the reality is that they are only two members of a much larger constellation of markers.

These potential pitfalls were avoided through use of the breath test in this study. First, subambient temperature programming was employed in the gas chromatograph. This provided superior separation of VOCs which might otherwise have co-eluted with one another, and the purity of every VOC in the BMAC in all subjects was confirmed by mass spectroscopy 14, 15. Secondly, for every subject studied, two samples (one of breath and one of room air) were routinely collected and assayed. This made it possible to determine the alveolar gradient of a VOC (abundance in breath‐abundance in room air), which varies with the difference between the rates of synthesis and clearance of a VOC in the body 14, 15. Thirdly, the BMAC was employed as a set of markers of oxidative stress. The advantage of the BMAC is that it employs the range of C4–C20 alkanes and their monomethylated derivatives as a composite set of markers of oxidative stress. In this set, pentane is only one VOC in a spectrum of 107 different VOCs, and the BMAC is less likely to be skewed by the relative increase or decrease of a single VOC.

Oxidative stress caused by oxygen toxicity may have adverse effects on virtually every organ of the body. This has been most extensively demonstrated in neonates, where oxygen toxicity has long been linked to retrolental fibroplasia and bronchopulmonary dysplasia 22. In adults, prolonged exposure to hyperbaric oxygen can cause central nervous system toxicity, atelectasis, pulmonary oedema and seizures 23. The adverse effects of normobaric hyperoxia are less well defined, probably because of the time lag between injuries sustained at the molecular level and subsequent pathological changes. However, increased oxidative stress has been shown to modify the structure of proteins 24 and DNA 25 and has been linked to an increased risk of cancer 26, 27 and chronic degenerative diseases, including atherosclerosis, heart failure, Parkinson's disease and Alzheimer's disease 27–30.

The authors conclude that breathing 28% oxygen for 30 min caused a significant increase in oxidative stress in normal volunteers. The clinical significance of this finding is not known, since a transient increase in oxidative stress may not necessarily progress to detectable cellular dysfunction or damage. However, in view of the well-documented adverse effects of prolonged oxidative stress, these findings reinforce what clinicians already know well: the routine use of oxygen to induce hyperoxia is potentially hazardous and it should be reserved only for situations in which it has demonstrated clinical benefit.

  • Received June 21, 2002.
  • Accepted August 14, 2002.
  • © ERS Journals Ltd

References

  1. ↵
    Knight JA. Free radicals: their history and current status in aging and disease. Ann Clin Lab Sci 1998;28:331–346.
    OpenUrlAbstract
  2. ↵
    Bostek CC. Oxygen toxicity: an introduction. AANA J 1989;57:231–237.
    OpenUrlPubMed
  3. ↵
    Kneepkens CM, Ferreira C, Lepage G, Roy CC. The hydrocarbon breath test in the study of lipid peroxidation: principles and practice. Clin Invest Med 1992;15:163–186.
    OpenUrlPubMedWeb of Science
  4. ↵
    Kneepkens CM, Lepage G, Roy CC. The potential of the hydrocarbon breath test as a measure of lipid peroxidation. Free Radic Biol Med 1994;17:127–160.
    OpenUrlCrossRefPubMedWeb of Science
  5. ↵
    Ambrosone CB. Oxidants and antioxidants in breast cancer. Antioxid Redox Signal 2000;2:903–917.
    OpenUrlPubMed
  6. ↵
    Onorato JM, Thorpe SR, Baynes JW. Immunohistochemical and ELISA assays for biomarkers of oxidative stress in aging and disease. Ann N Y Acad Sci 1998;854:277–290.
    OpenUrlCrossRefPubMedWeb of Science
  7. ↵
    Chandra M, Chandra N, Agrawal R, Kumar A, Ghatak A, Pandey VC. The free radical system in ischemic heart disease. Int J Cardiol 1994;43:121–125.
    OpenUrlCrossRefPubMedWeb of Science
  8. ↵
    Emelyanov A, Fedoseev G, Abulimity A, et al. Elevated concentrations of exhaled hydrogen peroxide in asthmatic patients. Chest 2001;120:1136–1139.
    OpenUrlCrossRefPubMedWeb of Science
  9. ↵
    Morita S, Snider MT, Inada Y. Increased N‐pentane excretion in humans: a consequence of pulmonary oxygen exposure. Anesthesiology 1986;64:730–733.
    OpenUrlPubMedWeb of Science
  10. ↵
    Loiseaux-Meunier MN, Bedu M, Gentou C, Pepin D, Coudert J, Caillaud D. Oxygen toxicity: simultaneous measure of pentane and malondialdehyde in humans exposed to hyperoxia. Biomed Pharmacother 2001;55:163–169.
    OpenUrlCrossRefPubMed
  11. Roberts RJ, Rendak I, Bucher JR. Lipid peroxidation in the newborn rat: influence of fasting and hyperoxia on ethane and pentane in expired air. Dev Pharmacol Ther 1983;6:170–178.
    OpenUrlPubMedWeb of Science
  12. ↵
    Turanlahti M, Pesonen E, Lassus P, Andersson S. Nitric oxide and hyperoxia in oxidative lung injury. Acta Paediatr 2000;89:966–970.
    OpenUrlCrossRefPubMedWeb of Science
  13. ↵
    Phillips M, Cataneo RN, Greenberg J, Gunawardena R, Naidu A, Rahbari-Oskoui F. Effect of age on the breath methylated alkane contour, a display of apparent new markers of oxidative stress. J Lab Clin Med 2000;136:243–249.
    OpenUrlCrossRefPubMedWeb of Science
  14. ↵
    Phillips M. Method for the collection and assay of volatile organic compounds in breath. Anal Biochem 1997;247:272–278.
    OpenUrlCrossRefPubMedWeb of Science
  15. ↵
    Phillips M, Herrera J, Krishnan S, Zain M, Greenberg J, Cataneo RN. Variation in volatile organic compounds in the breath of normal humans. J Chromatogr B Biomed Sci Appl 1999;729:75–88.
    OpenUrlCrossRefPubMed
  16. ↵
    Phillips M, Greenberg J, Sabas M. Alveolar gradient of pentane in normal human breath. Free Radic Res 1994;20:333–337.
    OpenUrlPubMedWeb of Science
  17. ↵
    Hansen TN, Smith CV, Gest AL, Smith HW, Giesler M. Biochemical manifestations of oxygen toxicity in the newborn lamb. Pediatr Res 1990;28:613–617.
    OpenUrlPubMedWeb of Science
  18. ↵
    Drury JA, Nycyk JA, Cooke RW. Pentane measurement in ventilated infants using a commercially available system. Free Radic Biol Med 1997;22:895–900.
    OpenUrlCrossRefPubMedWeb of Science
  19. ↵
    Nycyk JA, Drury JA, Cooke RW. Breath pentane as a marker for lipid peroxidation and adverse outcome in preterm infants. Arch Dis Child Fetal Neonatal Ed 1998;79:F67–F69.
    OpenUrlAbstract/FREE Full Text
  20. ↵
    Cailleux A, Allain P. Is pentane a normal constituent of human breath?. Free Radic Res Commun 1993;18:323–327.
    OpenUrlPubMedWeb of Science
  21. ↵
    Kohlmuller D, Kochen W. Is n‐pentane really an index of lipid peroxidation in humans and animals? A methodological reevaluation. Anal Biochem 1993;210:268–276.
    OpenUrlCrossRefPubMedWeb of Science
  22. ↵
    Saugstad OD. Chronic lung disease: oxygen dogma revisited. Acta Paediatr 2001;90:113–115.
    OpenUrlCrossRefPubMedWeb of Science
  23. ↵
    Jenkinson SG. Oxygen toxicity. New Horiz 1993;1:504–511.
    OpenUrlPubMed
  24. ↵
    Naskalski JW, Bartosz G. Oxidative modifications of protein structures. Adv Clin Chem 2000;35:161–253.
    OpenUrlPubMedWeb of Science
  25. ↵
    De Flora S, Izzotti A, Randerath K, et al. DNA adducts and chronic degenerative disease. Pathogenetic relevance and implications in preventive medicine. Mutat Res 1996;366:197–238.
    OpenUrlPubMedWeb of Science
  26. ↵
    Dreosti IE. Nutrition, cancer, and aging. Ann N Y Acad Sci 1998;854:371–377.
    OpenUrlCrossRefPubMedWeb of Science
  27. ↵
    Loft S, Poulsen HE. Cancer risk and oxidative DNA damage in man. J Mol Med 1996;74:297–312.
    OpenUrlCrossRefPubMedWeb of Science
  28. Hemnani T, Parihar MS. Reactive oxygen species and oxidative DNA damage. Indian J Physiol Pharmacol 1998;42:440–452.
    OpenUrlPubMed
  29. Gotz ME, Kunig G, Riederer P, Youdim MB. Oxidative stress: free radical production in neural degeneration. Pharmacol Ther 1994;63:37–122.
    OpenUrlCrossRefPubMedWeb of Science
  30. ↵
    Perry G, Smith MA. Is the lesion produced by oxidation a central part in the pathogenesis of Alzheimer's disease?. Neurologia 1999;14:78–84.
    OpenUrlPubMed
View Abstract
PreviousNext
Back to top
View this article with LENS
Vol 21 Issue 1 Table of Contents
  • Table of Contents
  • Index by author
Email

Thank you for your interest in spreading the word on European Respiratory Society .

NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We do not capture any email address.

Enter multiple addresses on separate lines or separate them with commas.
Effect of oxygen on breath markers of oxidative stress
(Your Name) has sent you a message from European Respiratory Society
(Your Name) thought you would like to see the European Respiratory Society web site.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Print
Citation Tools
Effect of oxygen on breath markers of oxidative stress
M. Phillips, R.N. Cataneo, J. Greenberg, R. Grodman, R. Gunawardena, A. Naidu
European Respiratory Journal Jan 2003, 21 (1) 48-51; DOI: 10.1183/09031936.02.00053402

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero

Share
Effect of oxygen on breath markers of oxidative stress
M. Phillips, R.N. Cataneo, J. Greenberg, R. Grodman, R. Gunawardena, A. Naidu
European Respiratory Journal Jan 2003, 21 (1) 48-51; DOI: 10.1183/09031936.02.00053402
del.icio.us logo Digg logo Reddit logo Technorati logo Twitter logo CiteULike logo Connotea logo Facebook logo Google logo Mendeley logo
Full Text (PDF)

Jump To

  • Article
    • Abstract
    • Materials and methods
    • Results
    • Discussion
    • References
  • Figures & Data
  • Info & Metrics
  • PDF
  • Tweet Widget
  • Facebook Like
  • Google Plus One

More in this TOC Section

  • Markers of activity in clinically recovered human leukocyte antigen-DR17-positive sarcoidosis patients
  • Airway surface pH in subjects with cystic fibrosis
  • Nasal nitric oxide measurements for the screening of primary ciliary dyskinesia
Show more Original Articles: Biomarkers of Lung Disease

Related Articles

Navigate

  • Home
  • Current issue
  • Archive

About the ERJ

  • Journal information
  • Editorial board
  • Press
  • Permissions and reprints
  • Advertising

The European Respiratory Society

  • Society home
  • myERS
  • Privacy policy
  • Accessibility

ERS publications

  • European Respiratory Journal
  • ERJ Open Research
  • European Respiratory Review
  • Breathe
  • ERS books online
  • ERS Bookshop

Help

  • Feedback

For authors

  • Instructions for authors
  • Publication ethics and malpractice
  • Submit a manuscript

For readers

  • Alerts
  • Subjects
  • Podcasts
  • RSS

Subscriptions

  • Accessing the ERS publications

Contact us

European Respiratory Society
442 Glossop Road
Sheffield S10 2PX
United Kingdom
Tel: +44 114 2672860
Email: journals@ersnet.org

ISSN

Print ISSN:  0903-1936
Online ISSN: 1399-3003

Copyright © 2023 by the European Respiratory Society