Using GC–MS to Track Effects of Second-Hand Cigarette Smoke in Nightclub Attendees

A recent pilot study conducted by the University of Split School of Medicine (Split, Croatia) established changes in the levels of nicotine and its metabolites in the urine as biomarkers of exposure to tobacco smoke in young 20–30 years old participants who attend nightclubs. Gas chromatography-mass spectrometry (GC–MS) was used to analyze the collected urine samples. An article based on this research was presented in the Croatian journal Arhiv za Higijenu Rada i Toksikologiju (Archives of Industrial Hygiene and Toxicology) (1).

According to the World Health Organization (WHO) estimates, there are 1.25 billion tobacco users in the world (2). Tobacco is associated with over eight million deaths a year, 1.3 million of which are owed to second-hand smoke (3). Second-hand or environmental tobacco smoke (ETS) is a mixture of smoke exhaled by an active smoker and the smoke released from a burning cigarette (4). In adults, second-hand smoke exposure has been associated with a 20–30 % higher risk of lung cancer in people living with a smoker and a 16–19 % higher risk in people exposed at the workplace (4,5) as well as a higher risk of cardiovascular diseases and stroke (6.7) Despite the overwhelming evidence of the harmful effects of tobacco smoke, second-hand exposure is still common and almost impossible to avoid in everyday social life. A 2001–2002 study that measured exposure to second-hand smoke at airports, train stations, hospitals, schools, universities, restaurants, and nightclubs across seven European cities found that it was present in most public places, with the highest nicotine concentrations measured in cafes and nightclubs (8).

This study included a random sample 22 participants from Split and Kutina, Croatia (15 women and seven men, of whom 17 non-smokers and five smokers [all of them women]) with an average age was 24 years. Urine samples were collected from the participants at two time points, with the first sample was collected before going to the nightclub, and the second after having spent 190 min in average in the nightclub and after six hours sleep. All samples (5–10 mL each) were stored at −20 °C until analysis, at which at time of analysis, 1.8 g of sodium tungstate dihydrate and 3 mL of dichloromethane/ethyl acetate mixture were inserted into a glass tube and 2 mL of urine was added. The content was mixed in a rotary mixer at 0.56 g for 10 min, and then centrifuged at 1500 g for another 10 min before evaporated to dryness under a stream of nitrogen. The dry residue was dissolved in 30 µL chloroform and transferred to a glass tube for a GC–MS run. The initial column temperature of 90 °C was held for 3 min, then ramped to 240 °C at 15 °C/min and held for the total run time of 13 min. Ultra-pure grade helium was used as a carrier gas at the flow rate of about 1.5 mL/min. One microliter of a sample was injected in splitless mode with an injection temperature of 250 °C. Ion source temperature was 200 °C, and the interface temperature was 280 °C. GC–MS analysis was performed using the selected ion monitoring mode with characteristic ions (m/z: 84, 133 and 162; 98, 176 and 42; 106, 192 and 135; for nicotine, cotinine and 3HC, respectively). Detector voltage was 1.5 kV in absolute mode (1).

The authors admit that their study and the interpretation of its findings are limited by the small number of participants and the fact that it could not be run under controlled conditions. Furthermore, to determine the real impact of passive smoking by excluding the contributions of active smoking, any further research should have two separate groups of participants (smokers and non-smokers). However, the authors state that the data compiled clearly shows a considerable increase in second-hand tobacco smoke exposure and a higher risk of its adverse effects in non-smokers attending nightclubs, as well as calls for further, more comprehensive research. Further investigations will include more participants and attention paid to variations in nicotine metabolism (1).

Group of men and women enjoying drinks at nightclub. © Jacob Lund - stock.adobe.com

References

1. Zečić, A.; Vazdar, B.; Slišković, L.; Sutlović, D. Urine Levels of Nicotine and its Metabolites in Young Population Exposed to Second-Hand Smoke in Nightclubs: A Pilot Study. Arh. Hig. Rad.a Toksikol. 2024, 75 (3), 211–216. DOI: 10.2478/aiht-2024-75-3846

2. Tobacco Use Declines Despite Tobacco Industry Efforts to Jeopardize Progress. World Health Organization.https://www.who.int/news/item/16-01-2024-tobacco-use-declines-despite-tobacco-industry-efforts-to-jeopardize-progress (accessed 2024-08-28)

3. World Health Organization. WHO Report on the Global Tobacco Epidemic, 2023: Protect People from Tobacco Smoke. World Health Organization, 2023.

4. U.S. Department of Health and Human Services. The Health Consequences of Involuntary Exposure to Tobacco Smoke: A Report of the Surgeon General – Executive Summary. Atlanta (GA): Centers for Disease Control and Prevention (US); 2006.

5. Sasco, A, J.; Secretan, M. B.; Straif, K. Tobacco Smoking and Cancer: A Brief Review of Recent Epidemiological Evidence. Lung Cancer 2004, 45, S2, S3–D9. DOI: 10.1016/j.lungcan.2004.07.998

6. Khoramdad, M.; Vahedian-azimi, A.; Karimi, L.; Rahimi-Bashar F,; Amini, H.; Sahebkar, A. Association Between Passive Smoking and Cardiovascular Disease: A Systematic Review and Meta-Analysis. IUBMB Life 2020, 72, 677–686. DOI: 10.1002/iub.2207

7. Oono, I. P.; MacKay, D. F.; Pell, J. P. Meta-Analysis of the Association Between Secondhand Smoke Exposure and Stroke. J. Public Health 2011, 33, 496–502. DOI: 10.1093/pubmed/fdr025

8. Nebot, M.; López, M. J.; Gorini, G.; Neuberger, M.; Axelsson, S.; Pilali, M. et al. Environmental Tobacco Smoke Exposure in Public Places of European Cities. Tob. Control 2005, 14, 60–63. DOI: 10.1136/tc.2004.008581