Gas chromatography-mass spectrometry study on composition of volatile organic compounds in exhaled breath of radiation-damaged rats_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

XIE Xiaoyun ^1,2 , TANG Jian ^1,2 , XIAO Bingkun ² , MIAO Xiaoyao ² , LI Zhiheng ² ,  HUANG Rongqing ^1,2

1. Guangdong Pharmaceutical University, Guangzhou, Guangdong 510006, P. R. China;
2. Institute of Radiational Chinese Medicine, Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100850, P. R. China;

Corresponding author：

HUANG Rongqing, Email: hrongqing@163.com

Keywords：

Radiation damage; exhaled breath; volatile organic compounds; differential metabolites

DOI：

10.7507/1671-6205.202311059

Video：

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Objective To explore composition of volatile organic compounds (VOCs) in exhaled breath of low-dose radiation-damaged Sprague-Dawely (SD) rats by thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS), and search for the differential metabolites of VOCs in the series of rats after radiation damage, and establish a noninvasive radiation damage detection method. Methods SD rats were randomly divided into five groups (a blank group, a 0.5-Gy group, a 1-Gy group, a 2-Gy group, and a 3-Gy group), with 8 rats in each group. A low-dose radiation injury model was established in rats. After the cobalt source radiation damage was performed, the body weight of rats was recorded, peripheral blood hematology was analyzed, and the exhaled breath of rats was collected on the 1^st, 5^th, 9^th and 13^th day. The composition of VOCs in the exhaled breath was analyzed by using the TD30-GC-MS technique, and multivariate statistical analyses were carried out to explore and obtain the differentiated metabolites after the radiation damage. Results After radiation damage, the rats showed a short-term decrease in body weight, peripheral blood and lung tissue sections were different, and the content of VOCs components in the exhaled breath of the damaged rats was significantly different from that of the rats in the blank group. Among them, four VOCs, acetophenone, nonanal, decanal and tetradecane increased, while heptane, chlorobenzene, paraxylene and m-dichlorobenzene decreased. Conclusions Through the GC-MS analysis of the exhaled breath of rats, eight components of VOCs in the exhaled breath of rats can be used as differential metabolites of radiation damage. This study lays a foundation for the establishment of a GC-MS analysis method for the components of VOCs in the exhaled breath of rats, as well as for the development of a nondestructive analytical assay for biological radiation damage.

Citation： XIE Xiaoyun, TANG Jian, XIAO Bingkun, MIAO Xiaoyao, LI Zhiheng, HUANG Rongqing. Gas chromatography-mass spectrometry study on composition of volatile organic compounds in exhaled breath of radiation-damaged rats. Chinese Journal of Respiratory and Critical Care Medicine, 2024, 23(5): 340-345. doi: 10.7507/1671-6205.202311059 Copy

1.	Upadhyay R, Bazan JG. Advances in radiotherapy for breast cancer. Surg Oncol Clin N Am, 2023, 32(3): 515-536.
2.	Zhou Z, Guan B, Xia H, et al. Particle radiotherapy in the era of radioimmunotherapy. Cancer Lett, 2023, 567: 216268.
3.	Bentzen S. Preventing or reducing late side effects of radiation therapy: radiobiology meets molecular pathology. Nature reviews. Cancer, 2006, 6(9): 702-713.
4.	Wang ZD, Zhang XQ, Du J, et al. Continuous cytogenetic follow-up, over 5 years, of three individuals accidentally irradiated by a cobalt-60 source. Mutat Res Genet Toxicol Environ Mutagen, 2015, 779: 1-4.
5.	Han L, Gao Y, Wang P, et al. Cytogenetic biodosimetry for radiation accidents in China. Radiat Med Prot, 2020, 1(3): 133-139.
6.	Popov TA. Human exhaled breath analysis. Ann Allergy Asthma Immunol, 2011, 106(6): 451-456.
7.	Miekisch W, Schubert JK, Noeldge-Schomburg GFE. Diagnostic potential of breath analysis - focus on volatile organic compounds. Clinica Chimica Acta, 2004, 347: 25-39.
8.	Buszewski B, Kesy M, Ligor T, et al. Human exhaled air analytics: Biomarkers of diseases. Biomed Chromatogr, 2007, 21: 553-566.
9.	Halbritter S, Fedrigo M, Höllriegl V, et al. Human breath gas analysis in the screening of gestational diabetes mellitus. Diabetes Technol Ther, 2012, 14(10): 917-925.
10.	Paleczek A, Rydosz A. Review of the algorithms used in exhaled breath analysis for the detection of diabetes. J Breath Res, 2022, 16(2).
11.	Thriumani R, Zakaria A, Hashim YZH, et al. A study on volatile organic compounds emitted by in-vitro lung cancer cultured cells using gas sensor array and SPMEGCMS. BMC Cancer, 2018, 18(1): 362.
12.	Gashimova EM, Temerdashev AZ, Porkhanov VA, et al. Comparative analysis of pre- and post-surgery exhaled breath profiles of volatile organic compounds of patients with lung cancer and benign tumors. J Anal Chem, 2022, 77(12): 1547-1552.
13.	Giovannini G, Haick H, Garoli D. Detecting COVID-19 from breath: a game changer for a big challenge. ACS Sens, 2021, 6(4): 1408-1417.
14.	Xue C, Xu X, Liu Z, et al. Intelligent COVID-19 screening platform based on breath analysis. J Breath Res, 2022, 17(1).
15.	李根平. 实验动物科学基本知识和基本技术. 北京: 中国农业大学出版社, 2011: 136.
16.	Caldeira M, Barros AS, Bilelo MJ, et al. Profiling allergic asthma volatile metabolic patterns using a headspace-solid phase microextraction/gas chromatography based methodology. J Chromatogr A, 2011, 1218(24): 3771-3780.
17.	Fuchs P, Loeseken C, Schubert JK, et al. Breath gas aldehydes as biomarkers of lung cancer. Int J Cancer, 2010, 126: 2663-2670.
18.	Shakyawar SK, Mishra NK, Vellichirammal NN, et al. A review of radiation-induced alterations of multi-omic profiles, radiation injury biomarkers, and countermeasures. Radiat Res, 2023, 199(1): 89-111.
19.	Haick H, Broza YY, Mochalski P, et al. Assessment, origin, and implementation of breath volatile cancer markers. Chem Soc Rev, 2014, 43(5): 1423-1449.
20.	Oh B, Figtree G, Costa D, et al. Oxidative stress in prostate cancer patients: a systematic review of case control studies. Prostate Int, 2016, 4: 71-87.
21.	Muzio G, Maggiora M, Paiuzzi E, et al. Aldehyde dehydrogenases and cell proliferation. Free Radic Biol Med. 2012, 52(4): 735-746.
22.	Leibman KC. Reduction of ketones in liver cytosol. Xenobiotica, 1971, 1(1): 97-104.
23.	Luo C, Sun J, Liu D, et al. Self-assembled redox dual-responsive prodrug-nanosystem formed by single thioether-bridged paclitaxel-fatty acid conjugate for cancer chemotherapy. Nano Lett, 2016, 16(9): 5401-5408.
24.	Phillips M, Cataneo RN, Greenberg J, et al. Effect of age on the breath methylated alkane contour, a display of apparent new markers of oxidative stress. J Lab Clin Med, 2000, 136(3): 243-249.
25.	Phillips M, Cataneo RN, Chaturvedi A, et al. Breath biomarkers of whole-body gamma irradiation in the Göttingen minipig. Health Phys, 2015, 108(5): 538-546.
26.	Simmons JE, Allis JW, Grose EC, et al. Assessment of the hepatotoxicity of acute and short-term exposure to inhaled p-xylene in F-344 rats. J Toxicol Environ Health, 1991, 32(3): 295-306.
27.	Roberts AE, Bogdanffy MS, Brown DR, et al. Lung metabolism of benzo[a]pyrene in rats treated with p‐xylene and/or ethanol. J Toxicol Environ Health, 1986, 18(2): 257-266.
28.	Patel JM, Wolf CR, Philpot RM. Interaction of 4-methylbenzaldehyde with rabbit pulmonary cytochrome P-450 in the intact animal, microsomes, and purified systems. Destructive and protective reactions. Biochem Pharmacol, 1979, 28(13): 2031-2036.
29.	庞硕, 吕丹, 张连峰. 二烯丙基硫醚通过靶向抑制CYP2E1对相关疾病治疗的潜在作用. 中国比较医学杂志, 2019, 29(8): 117-121.
30.	Selander HG, Jerina DM, Daly JW. Metobolism of chlorobenzene with hepatic microsomes and solubilized cytochrome P-450 systems. Arch Biochem Biophys, 1975, 168(1): 309-321.
31.	Korzekwa KR, Swinney DC, Trager WF. Isotopically labeled chlorobenzenes as probes for the mechanism of cytochrome P-450 catalyzed aromatic hydroxylation. Biochemistry, 1989, 28(23): 9019-9027.
32.	Krewet E, Müller G, Norpoth K. The excretion of chlorophenylmercapturic acid, chlorophenols and a guanine adduct in the urine of chlorobenzene-treated rats after phenobarbital pretreatment. Toxicology, 1989, 59(1): 67-79.
33.	Kimura R, Sano H, Itagaki K, et al. Identification of sulfur-containing metabolites of m-dichlorobenzene and their disposition and relationship with glutathione in rats. J Pharm Dyn, 1984, 7: 234-245.
34.	Kimura R, Ohishi N, Kato Y, et al. Identification of biliary metabolites of m-dichlorobenzene in rats. Drug Metab Dispos, 1992, 20: 161-171.
35.	Schettgen T, Bertram J, Krabbe J, et al. Excretion kinetics of 1, 3-dichlorobenzene and its urinary metabolites after controlled airborne exposure in human volunteers. Arch Toxicol, 2023, 97(4): 1033-1045.

1. Upadhyay R, Bazan JG. Advances in radiotherapy for breast cancer. Surg Oncol Clin N Am, 2023, 32(3): 515-536.
2. Zhou Z, Guan B, Xia H, et al. Particle radiotherapy in the era of radioimmunotherapy. Cancer Lett, 2023, 567: 216268.
3. Bentzen S. Preventing or reducing late side effects of radiation therapy: radiobiology meets molecular pathology. Nature reviews. Cancer, 2006, 6(9): 702-713.
4. Wang ZD, Zhang XQ, Du J, et al. Continuous cytogenetic follow-up, over 5 years, of three individuals accidentally irradiated by a cobalt-60 source. Mutat Res Genet Toxicol Environ Mutagen, 2015, 779: 1-4.
5. Han L, Gao Y, Wang P, et al. Cytogenetic biodosimetry for radiation accidents in China. Radiat Med Prot, 2020, 1(3): 133-139.
6. Popov TA. Human exhaled breath analysis. Ann Allergy Asthma Immunol, 2011, 106(6): 451-456.
7. Miekisch W, Schubert JK, Noeldge-Schomburg GFE. Diagnostic potential of breath analysis - focus on volatile organic compounds. Clinica Chimica Acta, 2004, 347: 25-39.
8. Buszewski B, Kesy M, Ligor T, et al. Human exhaled air analytics: Biomarkers of diseases. Biomed Chromatogr, 2007, 21: 553-566.
9. Halbritter S, Fedrigo M, Höllriegl V, et al. Human breath gas analysis in the screening of gestational diabetes mellitus. Diabetes Technol Ther, 2012, 14(10): 917-925.
10. Paleczek A, Rydosz A. Review of the algorithms used in exhaled breath analysis for the detection of diabetes. J Breath Res, 2022, 16(2).
11. Thriumani R, Zakaria A, Hashim YZH, et al. A study on volatile organic compounds emitted by in-vitro lung cancer cultured cells using gas sensor array and SPMEGCMS. BMC Cancer, 2018, 18(1): 362.
12. Gashimova EM, Temerdashev AZ, Porkhanov VA, et al. Comparative analysis of pre- and post-surgery exhaled breath profiles of volatile organic compounds of patients with lung cancer and benign tumors. J Anal Chem, 2022, 77(12): 1547-1552.
13. Giovannini G, Haick H, Garoli D. Detecting COVID-19 from breath: a game changer for a big challenge. ACS Sens, 2021, 6(4): 1408-1417.
14. Xue C, Xu X, Liu Z, et al. Intelligent COVID-19 screening platform based on breath analysis. J Breath Res, 2022, 17(1).
15. 李根平. 实验动物科学基本知识和基本技术. 北京: 中国农业大学出版社, 2011: 136.
16. Caldeira M, Barros AS, Bilelo MJ, et al. Profiling allergic asthma volatile metabolic patterns using a headspace-solid phase microextraction/gas chromatography based methodology. J Chromatogr A, 2011, 1218(24): 3771-3780.
17. Fuchs P, Loeseken C, Schubert JK, et al. Breath gas aldehydes as biomarkers of lung cancer. Int J Cancer, 2010, 126: 2663-2670.
18. Shakyawar SK, Mishra NK, Vellichirammal NN, et al. A review of radiation-induced alterations of multi-omic profiles, radiation injury biomarkers, and countermeasures. Radiat Res, 2023, 199(1): 89-111.
19. Haick H, Broza YY, Mochalski P, et al. Assessment, origin, and implementation of breath volatile cancer markers. Chem Soc Rev, 2014, 43(5): 1423-1449.
20. Oh B, Figtree G, Costa D, et al. Oxidative stress in prostate cancer patients: a systematic review of case control studies. Prostate Int, 2016, 4: 71-87.
21. Muzio G, Maggiora M, Paiuzzi E, et al. Aldehyde dehydrogenases and cell proliferation. Free Radic Biol Med. 2012, 52(4): 735-746.
22. Leibman KC. Reduction of ketones in liver cytosol. Xenobiotica, 1971, 1(1): 97-104.
23. Luo C, Sun J, Liu D, et al. Self-assembled redox dual-responsive prodrug-nanosystem formed by single thioether-bridged paclitaxel-fatty acid conjugate for cancer chemotherapy. Nano Lett, 2016, 16(9): 5401-5408.
24. Phillips M, Cataneo RN, Greenberg J, et al. Effect of age on the breath methylated alkane contour, a display of apparent new markers of oxidative stress. J Lab Clin Med, 2000, 136(3): 243-249.
25. Phillips M, Cataneo RN, Chaturvedi A, et al. Breath biomarkers of whole-body gamma irradiation in the Göttingen minipig. Health Phys, 2015, 108(5): 538-546.
26. Simmons JE, Allis JW, Grose EC, et al. Assessment of the hepatotoxicity of acute and short-term exposure to inhaled p-xylene in F-344 rats. J Toxicol Environ Health, 1991, 32(3): 295-306.
27. Roberts AE, Bogdanffy MS, Brown DR, et al. Lung metabolism of benzo[a]pyrene in rats treated with p‐xylene and/or ethanol. J Toxicol Environ Health, 1986, 18(2): 257-266.
28. Patel JM, Wolf CR, Philpot RM. Interaction of 4-methylbenzaldehyde with rabbit pulmonary cytochrome P-450 in the intact animal, microsomes, and purified systems. Destructive and protective reactions. Biochem Pharmacol, 1979, 28(13): 2031-2036.
29. 庞硕, 吕丹, 张连峰. 二烯丙基硫醚通过靶向抑制CYP2E1对相关疾病治疗的潜在作用. 中国比较医学杂志, 2019, 29(8): 117-121.
30. Selander HG, Jerina DM, Daly JW. Metobolism of chlorobenzene with hepatic microsomes and solubilized cytochrome P-450 systems. Arch Biochem Biophys, 1975, 168(1): 309-321.
31. Korzekwa KR, Swinney DC, Trager WF. Isotopically labeled chlorobenzenes as probes for the mechanism of cytochrome P-450 catalyzed aromatic hydroxylation. Biochemistry, 1989, 28(23): 9019-9027.
32. Krewet E, Müller G, Norpoth K. The excretion of chlorophenylmercapturic acid, chlorophenols and a guanine adduct in the urine of chlorobenzene-treated rats after phenobarbital pretreatment. Toxicology, 1989, 59(1): 67-79.
33. Kimura R, Sano H, Itagaki K, et al. Identification of sulfur-containing metabolites of m-dichlorobenzene and their disposition and relationship with glutathione in rats. J Pharm Dyn, 1984, 7: 234-245.
34. Kimura R, Ohishi N, Kato Y, et al. Identification of biliary metabolites of m-dichlorobenzene in rats. Drug Metab Dispos, 1992, 20: 161-171.
35. Schettgen T, Bertram J, Krabbe J, et al. Excretion kinetics of 1, 3-dichlorobenzene and its urinary metabolites after controlled airborne exposure in human volunteers. Arch Toxicol, 2023, 97(4): 1033-1045.

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Gas chromatography-mass spectrometry study on composition of volatile organic compounds in exhaled breath of radiation-damaged rats

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