Evidence of Limonene in Breath Samples in Men from the World’s Highest City

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Christian Sarbach Samuel Vergès Florine Coiffard Christophe Constancias Benoit Champigneulle Stéphane Doutreleau Philippe Andreucci Laurent Duraffourg Eric Postaire


A study to measure the composition of exhaled air was carried out in subjects living in the highest city in the world: La Rinconada in Peru.

People living at high altitude have a strongly disturbed metabolism and have to deal with major physiological disturbances. It is very likely that these subjects develop a disturbance of their redox system. It was therefore undertaken to study their ability to resist oxidative stress by measuring markers in the exhaled air. The traditional biomarkers encountered in this case are the alkanes resulting from the oxidative degradation of unsaturated fatty acids.

The breath was collected on carbotrap tubes thanks to the Exp'Air sampling system. The analyses were carried out by GC-MS. In addition to some alkanes, witnesses of the particular conditions of their exposure to the low quantities of oxygen present in the air, it was discovered surprisingly the presence of a compound rarely detected in the exhaled air, limonene. Hypotheses are presented but no certainty exists as to the origin of this compound in the breath. Further studies are underway to determine the conditions and circumstances of occurrence of limonene in the exhaled air of these subjects chronically exposed to hypoxia.       

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SARBACH, Christian et al. Evidence of Limonene in Breath Samples in Men from the World’s Highest City. Medical Research Archives, [S.l.], v. 10, n. 5, june 2022. ISSN 2375-1924. Available at: <https://esmed.org/MRA/mra/article/view/2798>. Date accessed: 30 june 2022. doi: https://doi.org/10.18103/mra.v10i5.2798.
Research Articles


Tremblay JC, Ainslie PN. Global and country-level estimates of human population at high altitude. Proc National Acad Sci 2021; 118(18): e2102463118.
West JB. Physiological Effects of Chronic Hypoxia. New England Journal of Medicine 2017; 376(20): 1965–1971
West JB. Physiological Effects of Chronic Hypoxia. New England Journal of Medicine 2017; 376(20): 1965–1971
Enserink M. Hypoxia city. Science 2019; 365(6458):1098–1103.
Beall CM. Two routes to functional adaptation: Tibetan and Andean high-altitude natives. Proceedings of the National Academy of Sciences of the United States of America 2007; 104 Suppl 1(Supplement 1): 8655–8660
Oberholzer L, Lundby C, Stauffer E, et al. Reevaluation of excessive erythrocytosis in diagnosing chronic mountain sickness in men from the world’s highest city. Blood 2020; 136(16): 1884–1888
Hancco I, Bailly S, Baillieul S, et al. Excessive Erythrocytosis and Chronic Mountain Sickness in Dwellers of the Highest City in the World. Front Physiol 2020; 11:773
León-Velarde F, Maggiorini M, Reeves JT, et al. Consensus Statement on Chronic and Subacute High Altitude Diseases. High Alt Med Biol 2005; 6(2):147–157
Stauffer E, Loyrion E, Hancco I, et al. What determines blood viscosity at the highest city in the world? J Physiology 2020; 598(18): 3817–3818
Villafuerte FC, Corante N. Chronic Mountain Sickness: Clinical Aspects, Etiology, Management, and Treatment. High Altitude Medicine & Biology [Internet] 2016; 17(2): 61–69.
Penaloza D, Arias-Stella J. The Heart and Pulmonary Circulation at High Altitudes. Circulation [Internet] 2007; 115(9): 1132–1146.
Bailey DM, Rimoldi SF, Rexhaj E, et al. Oxidative-Nitrosative Stress and Systemic Vascular Function in Highlanders With and Without Exaggerated Hypoxemia. Chest 2013; 143(2): 444–451
Pryor WA, The role of free radical reactions in biological systems. In: Pryor WA editor Free radicals biology. Vol. IV. New York: Academic Press, 1980; 1-49
Kohlmüller 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
Raspaud S, Postaire E, Massias L, et al. Evidence of oxidative burst in patients on chronic haemodialysis by alkane measurements. Frontiers of reactive oxygen species in biology and medicine, 1994; 457-458
Postaire E, Massias L, Lopez O, et al. Alkanes measurements in human immunodeficiency virus infection. In C. Pasquier [Ed.] Oxidative stress, Cell activation and viral infection. Birkhäuser Verlag Basel (Switzerland), 1994
Mendis S, Sobotka PA, Leja FL, Euler DE. Breath pentane and plasma lipid peroxides in ischemic heart disease. Free Radic Biol Med 1995; 20: 679-684
Olopade CO, Zakkar M, Swedler WI, et al.Exhaled pentane levels in acute asthma. CHEST 1997; Turanlahti M, Pesonen E, Lassus P, Anderson S. Nitric oxide and hyperoxia in oxidative lung injury. Acta Paediatr 2000; 89: 969- 970

Loiseaux-Meunier MN, Bedu M, Gentou C, et al. Oxygen toxicity: simultaneous measure of pentane and malondialdehyde in humans exposed to hyperoxia. Biomed Pharmacother 2001; 55: 163-169111: 862-865
Philipps M, Greenberg J, Cataneo RN. Effect of age on the profile of alkanes in normal breath. Free Radic Res 2000; 33: 57-63
Philipps M, Cataneo RN, Greenberg J, et al. Effect of oxygen on breath markers of oxidative stress. Eur Respir J 2003; 21: 48-51
Cataneo RN, Philipps M, Greenberg J, et al. Increased oxidative stress in younger as well as in older humans. Clin Chim Acta 2003; 328: 83-86
Mathews JM, Raymer JH, Velez GR, Garner E, et al. The influence of cytochrome P450 enzyme activity on the composition and quantity of volatile organics in expired breath. Biomarkers 1996; 1: 196-201
Mazzatenta A., Polorski M., Di Giulio C. Volatile organic compounds in exhaled breath as a marker of hypoxia in multiple chemical sensitivity. Physiological reports 2021; 9: e15035
O’Hara M E., Fernández del Río, Holt A., et al. Limonene in exhaled breath is elevated in hepatic encephalopathy. J.Breath Res. 2016 ; 10 : 046010
Arasaradnam R., Mcfarlane M., Ling K., Wurie S., et al. Breathomics—exhaled volatile organic compound analysis to detect hepatic encephalopathy: a pilot study J. Breath Res. 2016; 10: 016012
Luks AM, Auerbach PS, Freer L, et al. Wilderness Medical Society Practice Guidelines for the Prevention and Treatment of Acute Altitude Illness: 2019 Update. Wild Environ Med 2019;30(4):S3–S18
Wade CR, Van Rij AM. In vivo lipid peroxidation in man as measured by the respiratory excretin of ethane, pentane and other low-molecular-weight hydrocarbons. Anal Biochem 1985; 150: 1-7
Zarling EJ, Clapper M. Technique for gas-chromatographic measurement of volatile alkanes from single-breath samples. Clin Chem 1987; 33 (1): 140-141
Pleil JD, Linstrom AB. Exhaled human breath measurement method for assessing exposure to halogenated volatile organic coumpounds. Clin Chim Acta 2003; 28 (1-2): 83-86
Sarbach C., Stevens P., Whiting J., et al. Evidence of endogenous volatile organic compounds as biomarkers of diseases in alveolar breath. Ann. Pharmaceutiques Françaises, 2013 71 (4), 203-215
Sarbach C., Dugas B. and Postaire E. Evidence of variations of endogenous halogenated volatile organic compounds in alveolar breath after mental exercise-induced oxidative stress. Ann. Pharmaceutiques Françaises, 2020; 78 (1): 34-41
Sarbach C. and Postaire E. Biological markers of oxidative stress in exhaled air. Archives of Pharmacy and Pharmaceutical sciences, 2020; 4 (1): 10-12,. https://doi.org/10.29328/journal.apps.1001021
Delvordre P, Massias L, Sarbach C. Procédé et dispositif de recueil d’air expire aux fins d’analyse Patent n° 99 13582, 1999
Ferrandino G, Orf I, Smith R, et al.. Breath Biopsy Assessment of Liver Disease Using an Exogenous Volatile Organic Compound-Toward Improved Detection of Liver Impairment. Clin Transl Gastroenterol. Sep;11(9), 2020
Sinha R, Lockman KA, Homer NZM, et al. Volatomic analysis identifies compounds that can stratify non-alcoholic fatty liver disease. JHEP Rep. Jun 15;2(5):100137, 2020
Töreyin ZN, Ghosh M, Göksel Ö, et al. Exhaled Breath Analysis in Diagnosis of Malignant Pleural Mesothelioma: Systematic Review Int J Environ Res Public Health. Feb 10;17(3):1110, 2020

Fernández Del Río R, O'Hara ME, Holt A, Pemberton P, Shah T, Whitehouse T, Mayhew CA. Volatile Biomarkers in Breath Associated With Liver Cirrhosis - Comparisons of Pre- and Post-liver Transplant Breath Samples. EBioMedicine. Jul 26;2(9):1243-50, 2015
Anandakumar P, Kamaraj S, Vanitha MK. D-limonene: A multifunctional compound with potent therapeutic effects. D-limonene: A multifunctional compound with potent therapeutic effects. J Food Biochem. Jan;45(1), 2021
Mota-Gutierrez J, Barbosa-Pereira L, Ferrocino et al. Traceability of Functional Volatile Compounds Generated on Inoculated Cocoa Fermentation and Its Potential Health Benefits Nutrients. Apr 19;11(4):884, 2019
Hu Z; Li H; Weng Y; et al. Improve the production of d limonene by regulating the mevalonate pathway of Saccharomyces cerevisiae during alcoholic beverage. Fermentation Journal of Industrial Microbiology & Biotechnology 47:1083–1097, 2020