Analysis of Nicotine, Tar, CO, TPM, Moisture, BAP, and Humectants in Cigarettes and Beedis from India and Myanmar

Analysis of Nicotine, Tar, CO, TPM, Moisture, BAP, and Humectants in Cigarettes and Beedis from India and Myanmar | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Analysis of Nicotine, Tar, CO, TPM, Moisture, BAP, and Humectants in Cigarettes and Beedis from India and Myanmar Priyamvada Sharma, Jagdish Kaur, Arvind Vashishta Rinkoo, Vijayashree Rao, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6219017/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 30 Jan, 2026 Read the published version in Scientific Reports → Version 1 posted 11 You are reading this latest preprint version Abstract Approximately 2.3 million annual deaths in the eleven countries of the WHO Southeast Asia Region (SEAR) are linked to tobacco smoking. In 2020, smoking was responsible for 1.6 million lives lost in SEAR. The toxic substances found in the emissions of smoked tobacco products are inadequately researched. This study aims to evaluate and compare the smoke delivery potential of nicotine, tar, carbon monoxide, humectants, tobacco particulate matter (TPM), benzo[a]pyrene, and moisture between traditional beedis and commercial cigarettes. For quantifying emission ingredients from both beedis and cigarettes, globally accepted procedures from TobLabNet, the CDC, and CORESTA were utilized. When comparing Indian and Myanmar cigarettes, we discovered that nicotine and carbon monoxide levels in Myanmar cigarettes were slightly higher than those in Indian cigarettes, though the difference was statistically insignificant. Moisture, tar, and TPM also exhibited statistically insignificant variations. The mean benzo[a]pyrene levels in Indian and Myanmar products were 8.24 ± 0.28 and 12.17 ± 2.24, respectively, with a statistically significant difference (≤ 0.008). Among humectants, only propylene glycol showed significant variation (p ≤ 0.023). The comparison between Indian cigarettes and beedis revealed statistically significant differences in nicotine (p ≤ 0.041), moisture (p ≤ 0.001), TPM (p ≤ 0.001), propylene glycol (p ≤ 0.002), and glycerol (p ≤ 0.001). The mean benzo[a]pyrene for cigarettes was 8.24 ± 0.28, while for beedis, it was 10.22 ± 1.07, demonstrating a statistically significant difference (p ≤ 0.008), along with glycerol (p ≤ 0.009) among humectants. When comparing beedis to cigarettes from both countries, we found significant differences in nicotine (p ≤ 0.041), moisture (p ≤ 0.001), TPM (p ≤ 0.001), propylene glycol (p ≤ 0.002), and glycerol (p ≤ 0.001). Flavors were not detected in the mainstream smoke of the cigarettes and beedis analyzed for this study. The findings of this study can be leveraged to enhance public health by identifying harmful chemicals that exceed established limits and potentially motivating manufacturers to produce less harmful products by conforming to toxin emission standards. Health sciences/Health care Physical sciences/Chemistry Nicotine Tar Carbon Monoxide TPM Benzo[A]Pyrene Moisture Humectants Flavors cigarettes Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 What this paper adds Cigarette smoking remains a global source of major health problems and is the leading cause of preventable deaths and diseases due to non-communicable diseases globally. This research aimed to examine the mainstream smoke deliveries from popular cigarettes and beedis from India and Myanmar. The smoke deliveries of Nicotine, CO, moisture, and tar in mainstream smoke from 11 cigarettes and 9 beedi brands were analyzed. Humectants (Propylene glycol, glycerol, and triethylene glycol) and benzo [A] pyrenes were also tested. The level of nicotine in the Indian brands was lower than in the Myanmar brands. However, the locally produced Indian beedis showed higher nicotine, moisture, CO, and tar levels compared to cigarettes. Introduction Tobacco use, including smoking, leads to over 8 million deaths annually, including over 7 million from direct tobacco use and roughly 1.2 million from exposure to passive smoking. 1 . On average, smoking shortens life expectancy by about 10 years 2 . Tobacco use is a major threat to public health, it kills around 4 million people every year in the WHO Southeast Asia Region 1 . As per the fifth edition of the WHO global report on trends in the prevalence of tobacco use 2000–2030, the SEARO is home to around 199 million tobacco smokers (20% of the global total) and 280 million smokeless tobacco users (77% of the global total). Tobacco is consumed in diverse forms, including smoked products, such as cigarettes and beedis (a cheaper hand-rolled form of smoked tobacco), and a variety of smokeless tobacco (SLT). Comprehensive awareness of tobacco’s harmful effects is far from desirable among users due to culture-related customs and misbeliefs about their health effects 3 . All forms of tobacco are harmful, and there is no safe level of exposure to tobacco. Cigarette smoking is the most common form of tobacco use worldwide. Other smoked tobacco products include waterpipe, cigars, cigarillos, heated tobacco products (HTPs), roll-your-own tobacco, pipe tobacco, beedis, and kreteks. Around 80% of the 1.3 billion tobacco users worldwide live in low and middle-income countries 4 , where the burden of tobacco-related illness and death is heaviest. Tobacco use contributes to poverty by diverting household spending from basic needs such as food, education, and shelter to buying tobacco products. India is the second largest consumer and producer of tobacco in the world. The Global Adult Tobacco Survey 2017 revealed that 28.6% (around 266 million) of adults in India, aged 15 and above, currently use tobacco in some form. Of these, 10.7% (around 99 million) currently smoke tobacco and 21.4% (around 199 million) use smokeless tobacco 1 . Tobacco products increase the risk of heart and lung diseases, as well as cancers and a variety of other adverse health outcomes 3 – 4 . India's GDP is negatively impacted by tobacco use, and the direct medical expenses incurred in treating tobacco-related illnesses account for 5.3% of the nation's yearly spending on both public and private health care 5 . It is estimated that the economic costs of tobacco-related diseases are ten times higher than the amount of money India receives from tobacco taxes 5 . This drains the public health system and the economy, which no country can afford. In the Southeast Asian region, Myanmar reports the highest tobacco use, with over half of adults (54%) using some form of tobacco. STEPS 2014 further revealed that 26% of adults in Myanmar were tobacco smokers and 43% used smokeless tobacco 2 , 3 . India and Myanmar can avert millions of preventable deaths as well as mitigate the damage of tobacco use on both society and the economy through the implementation of a comprehensive package of strong tobacco control policies outlined in the WHO Framework Convention on Tobacco Control (FCTC). Nicotine in tobacco is responsible for its addictive nature. Nicotine is a paradoxical substance because it has both stimulating and depressant effects. It affects the cardiovascular, skeletal, gastrointestinal, and peripheral nervous systems, among other systems 2 . At the molecular levels, nicotine in tobacco mimics the neurotransmitter acetylcholine. Some of the effects of nicotine may be traced to its many receptors in cholinergic binding sites in the brain. Nicotine could be the main psychoactive component of tobacco due to its intricate pharmacological actions 6 . Several additional essences are purposefully added to tobacco products to make them attractive and to lessen their harshness and potential for dependence 2 . Many non-tobacco substances, such as humectants, fragrances, and flavors are added to attract and appeal to consumers. The American Lung Association has released data indicating that there are roughly 600 ingredients in one cigarette. Almost 7,000 chemicals are released when cigarettes burn, many of which are toxic. At least 69 of them are known to cause cancer 7 . To ensure compliance with WHO FCTC Article 9 and effective regulatory controls, the contents and emissions of tobacco products must be monitored. The WHO Tobacco Free Initiative (TFI) established the tobacco testing laboratory network (TobLabNet) in 2005 as part of its global agenda. This is based on the WHO’s Study Group on Tobacco Product Regulation (TobReg) recommendation to build and strengthen tobacco product testing and research capacity by pursuing Articles 9 and 10 of the WHO Framework Convention on Tobacco Control (WHO FCTC). Under WHO TFI's leadership, TobLabNet develops and validates methods and standard operating procedures (SOPs) for testing the contents and emissions of tobacco products. In India, 29% of all adults use tobacco, while in Myanmar, over half of adults (54.4%) use some form of tobacco 2 , 4 . A wide variety of smoked tobacco products are available in India and Myanmar. However, very little data on the emissions and addictiveness of smoked tobacco products is available from the region because of the required infrastructure and technical expertise involved in emission testing. This research aims to collect scientific information about the amount of nicotine, moisture, humectants, and benzo [A] pyrene in tobacco emissions using standard and validated methods and available laboratory support. Material and Methods This study was conducted from June 2023 to May 2024 at the Tobacco Testing and Drug Toxicology Laboratory, Centre for Addiction Medicine (CAM), National Institute of Mental Health and Neurosciences (NIMHANS), Bengaluru, Karnataka. The state-of-the-art Drug Toxicology Laboratory is a world-class facility with high-end and sophisticated drug and tobacco testing instruments. The Tobacco Testing facility is a member of the World Health Organization (WHO) global network of Tobacco Testing Laboratories. All the chemicals used for analysis were of analytical grade. Nicotine, n-Heptadecane (Internal standard), Benzo[A]Pyrene and Humectants (Glycerol, Propylene Glycol, Triethylene Glycol), 1, 3- Butanediol (Internal standard), Flavors (Methyl Salicylate, Ethyl Salicylate, Eugenol, Di-phenyl ether, Cinnamaldehyde, Menthol, Coumarin, Camphor), 3′, 4′-Methylenedioxy acetophenone (MDA) (Internal standard) and Solvents (Cyclohexane, Propan-2-ol, Benzene, Methanol) procured through Sigma Aldrich, USA having purity of ≥ 99.0%. 2.1 Patients and Public Involvement : Patients were not involved in this research. 2.2 Sample Collection and Storage: Samples were randomly selected from the cigarette sales points. Four cigarettes and nine Beedi brand samples were procured locally from wholesale and retail outlets in and around Bangalore, India. Seven cigarette brands sourced from Myanmar were obtained through WHO, SEARO, New Delhi, India. Reference cigarettes (3R4F, 1R5F, 2R4F) were obtained from the University of Kentucky. CORESTA Monitor (CM4 and CM6) reference cigarettes were received from CORESTA (Paris, France). The samples were transported in air-tight sealed packs to the Tobacco Testing Facility, NIMHANS, Bengaluru. The labeled samples were stored in sealed packs within 10 days of receipt at – 20°C (Ultra Low deep freezer from Vestfrost) in their original packaging following the International Organization for Standardization (ISO, Geneva, Switzerland) guidance document ISO 3402:1999). Before analysis, samples were refrigerated for 24 h for comprehensive re-equilibration, followed by 2 h of equilibration to ambient conditions in the Environmental Chamber, Thermo Scientific 3900 Series 8 . 2.3 Ethics approval and consent to participate Ethical approval was not required as this work did not involve human subjects. 3.1 Instrumentation For accurate testing of smoking behavior, it is crucial to replicate human smoking patterns using a smoke machine. This machine simulates human smoking by controlling the frequency and depth of inhalation, allowing researchers to analyze various factors such as chemical concentration, consistency, and total quantity. The Cerulean SM450i is a 20-channel semi-automatic linear smoking machine designed to replicate human smoking behavior and complies with the International Organization for Standardization (ISO 3308) standards, ensuring reliable results and regulatory compliance. It can handle a diverse range of cigarettes/beedis without sacrificing efficiency or throughput. Beedi production in India is unregulated and lacks automation. The tendu leaves used for wrapping are less combustible and nonporous, stronger puffs with greater volume and frequency may be required to ignite and maintain a consistent burn. The ISO 3308 standards may not be suitable for studying beedi emissions a relevant instrument and approach are necessary for accurate beedis analysis. ISO 17175 was preferred for beedis emission analysis. The sampling procedure entails puffing for two seconds at 30-second intervals. The Cerulean beedi holder (Part: 99301) and latex beedi sleeve (Part: KU0358) were used in conjunction with a handheld vacuum pump to smoke beedis. The emissions were collected in bags, and the carbon monoxide (CO) percentage by volume was measured using a non-dispersive infrared (NDIR) analyzer. Total Particulate Matter (TPM) was calculated as per ISO 3308 by weighing the holders before and after the smoke run 9 . The particulate matter collected on Cambridge filter pads (44 mm) was extracted as per TobLabNet SOP-10 for nicotine quantification. Nicotine quantification was done on an Agilent 8890 Gas Chromatograph with a Flame ionization detector. The chromatographic separation was achieved on CP-WAX 51, column (CP7405) in isothermal mode at 170º C shown in Figure 1. Humectants such as glycerol, propylene glycol, and triethylene glycol are added to tobacco products to facilitate the processing of the cured tobacco leaf, retain moisture, and increase shelf life. They dilute nicotine harshness and improve the sensory properties of tobacco products. Humectant quantification was done by TobLabNet SOP 6 using Agilent 8890 GC with FID. The humectants were separated on DB Wax (123-7032) fused silica column shown in Figure 2. Benzo[A]Pyrene belongs to the group of compounds known as polycyclic aromatic hydrocarbons (PAHs). PAHs are produced by heating or incomplete burning of organic material. They are not present in tobacco but are mostly produced as they burn. Exposure to BAP while smoking is one of the most important causes of cancer. Benzo[A]Pyrenes were quantified using TobLabNet SOP 5 on 5975 MSD clubbed with Agilent 7890 GC-MSD system. Chromatographic separation was achieved on the DB-5MS column (122-5532G). The mass spectra of ions produced by electro-spraying was identified by the National Institute of Standards and Technology (NIST) library and shown in Figure 3. 3.2 Statistical Analysis SPSS Statistics for Windows, version 23.0. 2015 (SPSS Inc., Chicago, Ill., USA) was used to perform the statistical analysis of the data. A comparison between cigarettes and beedis was done through non-parametric analysis. Mann–Whitney U-test was used for data with P < 0.05 was considered significant . Results The comparisons of nicotine, CO, moisture, and tar between the cigarette samples received from Myanmar and India are summarised in Figure 4. The mean nicotine and CO in Myanmar cigarettes was 0.84 mg/cig (±0.17), and 13.35 mg/cig (±3.90) respectively. In Indian cigarettes, the mean nicotine was 0.66 mg/cig ±0.08, and CO was 11.01 mg/cig ±1.14. Although the nicotine and CO levels in Myanmar cigarettes were slightly higher than in Indian cigarettes, the difference was not statistically significant (Figure 4). The mean moisture content in Indian cigarettes was 4.44 ±0.74, while for Myanmar cigarettes, it was 4.17 ±1.93 but the difference was statistically insignificant. The mean Tar and TPM in the Indian products were 10.34 ± 2.03 mg/cig, and 15.44 ± 2.13 mg/cig respectively, while the Myanmar products had a mean Tar 12.02± 4.05 and TPM 17.03 ± 2.13 mg/cig however, the difference was not statistically significant (Figure 4). The mean Benzo[A]Pyrene in Indian cigarettes was 8.24 ± 0.28, while the cigarettes from Myanmar cigarettes reported high BAP levels with a mean of 12.17± 2.24. A statistically significant (≤0.008) variation was observed in BAP levels (Figure 5). Humectants are added to cigarettes to control and maintain the moisture content of the tobacco filler. The mean glycerol levels in Indian cigarettes were 10.27 mg/gm ± 5.65, while the mean propylene glycol was 0.38 mg/gm ± 0.30 and triethylene glycol was 0.03 mg/gm ± 0.04. Myanmar cigarettes had mean glycerol levels of 10.02 mg/gm ± 5.08, propylene glycol, 2.52 mg/gm ± 1.32, and triethylene glycol 0.02 mg/gm ± 0.06. On comparing all three humectants, we found there was no statistically significant variation in the glycerol and triethylene glycol, but propylene glycol was statistically significant (p ≤ 0.023) (Figure 5). Beedi had mean nicotine levels of 1.36 mg/beedi ±0.56 and mean CO levels of 13.95 mg/beedi ±2.43 (Figure 6). The moisture content of beedis varied from 1.32 to 28.4 mg/beedi, with a mean of 18.27 mg/beedi ± 5.95. The mean Tar levels were 19.22 ± 11.94 and TPM levels were 38.85 mg/beedi ± 7.80. On comparing Indian Cigarettes with Beedis we report Nicotine, CO (p ≤0.02), moisture, and TPM (≤0.008 ) were statistically significant. When Beedis were compared with cigarettes from both countries we found that nicotine (p ≤0.041), moisture (p ≤0.001), TPM (p ≤0.001), propylene glycol (p ≤0.002), and glycerol (p ≤0.001) differ significantly. Beedis had mean glycerol levels 0.91mg/gm ± 0.58, propylene glycol was 0.09mg/gm ± 0.06, and triethylene glycol levels 0.04 mg/gm ±0.03. On comparing beedi humectants with cigarettes, we found that values were statistically significant for glycerol (p ≤0.009) only (Figure 7). BAP for cigarettes had a mean of 8.24±0.28 while for beedis it was 10.22 ±1.07, the values were statistically significant (p ≤0.008). All cigarettes and beedis emissions were tested for flavors (Methyl Salicylate, Ethyl Salicylate, Eugenol, Eucalyptol, Di phenyl Ether, Cinnamaldehyde, Menthol, Coumarin, and Camphor) using Centre for Disease Control and Prevention (CDC) SOP, 60. Flavors were not detected in the samples received from both countries. Discussion Cigarettes are expertly crafted, mass-produced items available worldwide. They are filled with processed tobacco and wrapped in paper. Beedis are small, hand-rolled tobacco products made of sun-dried and processed tobacco wrapped in Tendu or Temburni leaves and secured with cotton thread 9 . India has the world's second-largest number of adult smokers. Beedi is the most commonly smoked tobacco product smoked by an estimated 71.8 million adults 2,4 . Tobacco smoke is a complex mixture of over seven thousand volatile and semi-volatile compounds. Particulate matter (PM) generated due to tobacco smoking can be coarse (≤10 µm), fine (≤2.5 µm), or ultrafine (≤1 µm) 11 . The size of PM indicates both its ability to adsorb toxic organic compounds and the depth to which it can enter the respiratory system. The protonated form of nicotine gets deposited on the PM and gradually absorbed by the body. The other compounds of tobacco smoke are humectants, volatile aldehydes, nitrogen oxides, and polyacrylic aromatic hydrocarbons. Many harmful and potential carcinogens are present in mainstream smoke. If inhaled, high levels of carbon monoxide, especially from beedis can enter the bloodstream and bind with hemoglobin to form carboxy-hemoglobin. This results in tissue ischemia, which is a major cause of cardiovascular diseases, common among beedi smokers 10 . The Government of India signed the Framework Convention on Tobacco Control (FCTC) (in 2004) and enacted the Cigarettes and Other Tobacco Products Act (COTPA) in 2003. The COTPA has not imposed any specific limitations on emissions or tobacco content. In its advisory note on the Global Nicotine Reduction Strategy, the WHO Study Group on Tobacco Product Regulation (TobReg) stated that the risk of dependence on cigarettes can be reduced by dropping their nicotine content to a very low level. The nicotine content should therefore be as low as is technically feasible. According to WHO's worldwide scan of national laws pertaining to maximum emission levels of tar, nicotine, and carbon dioxide (TNCO), as of December 31, 2022, eighty-three, nations had established maximum levels of nicotine (N) permitted per cigarette, with sixty of these nations (72%) allowing one milligram of nicotine per cigarette. Furthermore, sixty-seven countries permit ten milligrams of tar per cigarette, while eighty-five countries have set maximum levels of tar (T) per cigarette. Additionally, fifty-seven nations have set a carbon monoxide restriction of ten milligrams/ cigarette worldwide. In the Southeast Asian region, only Myanmar expressly forbids the exhibition of emission levels, At the same time Timor-Leste has established limits of ten milligrams for tar, one milligram for nicotine, and ten milligrams for carbon monoxide per cigarette (TNCO of 10/1/10 mg/cigarette). All the cigarette brands tested had nicotine levels (0.77 mg/cigarette ±0.16) within acceptable limits. For the beedis, the mean nicotine levels 1.36 mg/beedi ± 0.56 were above the maximum set limit (1.0mg). Ten milligrams of carbon monoxide per cig is the standard maximum permissible limit. Cigarette samples (n=6) received from Myanmar were found to exceed the common maximum allowable limit of 10mg carbon monoxide/cig. All beedis samples had CO levels above the set limit, the mean was 13.95 mg/beedi ±2.43. Beedis smokers are potentially exposed to significantly higher concentrations of nicotine due to the greater puffing frequency. Research has shown that the non-porous nature and higher moisture content of tendu leaf in beedis led to higher levels of carbon monoxide and tar in its smoke compared to regular cigarette smoke. The presence of harmful and carcinogenic chemicals in mainstream beedi smoke could be harmful to human health. 11 The Myanmar cigarettes had Tar levels ≥12.02mg of Tar/cig ±4.05, which was well above the highest limit (10.00mg/cig) set by countries. Indian beedis also had mean Tar levels well above the accepted limit (19.22 mg /beedi±11.94 ). Humectants including propylene glycol and glycerol were present in all the samples from both countries while trimethylene glycol was not detected in a few samples. Indian cigarettes and beedis had propylene glycol levels ≤ 1.0 mg/gm, and glycerol in Indian brands was ≥10 mg/gm except for one make which had very low glycerol levels (1.99 mg/gm). Triertylene glycol in Indian cigarettes was not detected in all the samples, only two samples had it in the range of 0.04 - 0.09 mg/gm. Yan Xizheg tested twenty-seven popular cigarette brands in the USA for humectants. Only four brands showed no detectable amounts of the humectants, and the rest of the brands showed the presence of glycerol or 1,2-propylene glycol with concentrations ranging from 1.66 - 3.57% and from 0.23 to 1.35% for glycerol and 1,2-propylene glycol, respectively. In general, all the manufacturers use more glycerol than 1,2-propylene glycol as a humectant agent which may serve as a precursor for the formation of harmful carbonyl compounds 12 . Benzo(a)pyrene (BAP) is a marker of carcinogenic activity of polycyclic aromatic hydrocarbon (PAH). PAH is present in emission exhausts in small quantities of cigarette smoke, typically <10 ng/cigarette. In Indian brands of cigarettes, BAP was ≤ 10 ng/cig. In the cigarettes received from Myanmar only two brands had BAP levels 9.95 and 9.72 ng/cig. In the rest of the samples, it was ≥ 10.0 ng/cig with a mean of 10.22 ng/beedi±1.07 which may be harmful to the users depending upon the consistency of use. Information regarding BAP levels in the products offered by SEARO is limited. The analysis of mainstream smoke of cigarettes from Nigeria revealed that the BAP levels varied between 0 and 22.7 ng/cig. The most prevalent PAH in all of the products examined was naphthalene, which ranged from 210.7 to 460.34 ng/ cig 13 . Conclusions Cigarettes and beedis present a significant challenge to the public’s health. Their availability and allure could override the health risks and negative perceptions attached to them. A quantitative analysis of emissions in mainstream smoke is necessary to assess the harmful effects of smoking. Current research was planned to quantitatively analyze Nicotine, CO, Moisture, Tar, TPM, humectants, and BAP, in the mainstream smoke of selected cigarette and beedi samples from India and Myanmar. Some popular brands of cigarettes (India and Myanmar) and beedis were analyzed using globally accepted SOPs. of ISO, CORESTA, and TobLabNet. We report statistically significant differences in BAP (≤0.008) and Triethylene glycol (≤0.023) values, between Indian and Myanmar cigarettes. On comparing Indian cigarettes with beedis nicotine, CO (≤0.023) and glycerol (≤0.001) showed a statistically significant difference. The contents and emissions were measured using globally approved protocols. This information can be further utilized to make policy decisions, educate the public about the dangers of smoking, and potentially incentivize tobacco companies to produce less harmful products by setting standards for emission levels of toxins. Limitation: Analysis of tobacco-specific nitrosamines could not be planned in the absence of reference standards. Abbreviations B[A]P: Benzo[A]Pyrene CAM: Centre for Addiction Medicine CDC: Centre for Disease Control and Prevention CFP: Cambridge Filter Pad FID: Flame Ionisation Detector GCMS: Gas Chromatography-Mass Spectrometry ISO: International Standard Organisation NDIR: Non- dispersive infrared Analyser NIMHANS: National Institute of Mental Health and Neurosciences TFI: Tobacco Free Initiative SOP: Standard Operating Procedure WHO: World Health Organisation SEARO: South East Asia Regional Office TobLabNet: Tobacco Laboratory Network Declarations Authors and Contributions: Prof. Pratima Murthy and Dr. Jagdish Kaur conceptualized this idea. Dr. Priyamvada Sharma wrote the primary manuscript. Ranti F helped writing and interpretation of results. Dr. Arvind V Rinkoo helped in the final editing of the manuscript. Prof. P Marimutthu performed the statistical analysis. Ms. Amina Salam and Ms. Vijayahree Rao conducted all the experimental work at NIMHANS, Bangalore, India. Competing interests : None declared Funding: This project was funded by WHO under the grant WHO Registration: 2023/1365732-0. PO: 2023166156 Data sharing statement: The datasets used and/or analysed during the current study are available from the corresponding author upon reasonable request. Disclaimer: The content is solely the responsibility of the authors and does not necessarily represent the views of the Institutes involved. References Kaur, J., Rinkoo, A. V. & Richardson, S. Update on numbers of tobacco-attributable deaths by country in the South-East Asia region: policy implications. Tob Control tc-2024-058599 (2024) doi:10.1136/tc-2024-058599. (Global Burden of Disease [database. Washington, DC: Institute of Health Metrics; 2019. IHME, accessed 17 July 2023). Chen, D. T.-H. et al. A longitudinal study of transitions between smoking and smokeless tobacco use from the ITC Bangladesh Surveys: implications for tobacco control in the Southeast Asia region. 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BMC Chemistry 13 , 93 (2019). Xizheng, Y., Valentín-Blasini, L., Watson, C. & Cardenas, R. B. Determination of Humectants in Tobacco Filler by High Performance Chromatography/Single Quadrupole Mass Spectrometry. Beiträge zur Tabakforschung International/Contributions to Tobacco Research 28 , 170–178 (2018). Adesina, O. A., Olowolafe, T. I. & Igbafe, A. Levels of polycyclic aromatic hydrocarbon from mainstream smoke of tobacco products and its risks assessment. Journal of Hazardous Materials Advances 5 , 100053 (2022). Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6219017","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":453365993,"identity":"88f54f86-b761-443e-a70d-1fc132c9ef82","order_by":0,"name":"Priyamvada Sharma","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA2klEQVRIiWNgGAWjYFAC5gMGHwwkePhB7IQCorSwJRTOqLCQk2wAaTEgSguPwWeeMxXGBgdAHGK0mPMfS9zA2yaRuPn86sQPDwwY5PnFDuDXYjkj+bCBJFDLthtvN0sAHWY4c3YCfi0GN9jSDAzBWs5uAGlJMLhNSMv5M+Y/EkEOm3F28w/itBzIMTA4cEbC2IC/dxtxtljOSEswbKiQkJO4wbvNIsFAgrBfzPkPHzD+Y1DHw99/dvPNHxU28vzShBwGZ0mAVUrgV46qhf8AYdWjYBSMglEwMgEARG1IQfFHIwsAAAAASUVORK5CYII=","orcid":"","institution":"National Institute of Mental Health and Neurosciences (NIMHANS)","correspondingAuthor":true,"prefix":"","firstName":"Priyamvada","middleName":"","lastName":"Sharma","suffix":""},{"id":453365994,"identity":"aaf73fc6-6f10-4d32-b7d7-b50f3f5205e0","order_by":1,"name":"Jagdish Kaur","email":"","orcid":"","institution":"WHO Regional Office for South-East Asia","correspondingAuthor":false,"prefix":"","firstName":"Jagdish","middleName":"","lastName":"Kaur","suffix":""},{"id":453365995,"identity":"aeaeacb9-5f2c-4fd1-b0f3-5986f2992cfe","order_by":2,"name":"Arvind Vashishta Rinkoo","email":"","orcid":"","institution":"WHO Regional Office for South-East Asia","correspondingAuthor":false,"prefix":"","firstName":"Arvind","middleName":"Vashishta","lastName":"Rinkoo","suffix":""},{"id":453365996,"identity":"a36df178-54aa-4db3-ab0e-68bfd35b3e9a","order_by":3,"name":"Vijayashree Rao","email":"","orcid":"","institution":"National Institute of Mental Health and Neurosciences (NIMHANS)","correspondingAuthor":false,"prefix":"","firstName":"Vijayashree","middleName":"","lastName":"Rao","suffix":""},{"id":453365997,"identity":"de2a31e2-0b5f-4805-ac2b-8352776f0236","order_by":4,"name":"Amina Salam","email":"","orcid":"","institution":"National Institute of Mental Health and Neurosciences (NIMHANS)","correspondingAuthor":false,"prefix":"","firstName":"Amina","middleName":"","lastName":"Salam","suffix":""},{"id":453365998,"identity":"6036af9e-c9d0-44b7-94db-4eb7114f6b5e","order_by":5,"name":"Fayokun Ranti","email":"","orcid":"","institution":"World Health Organization","correspondingAuthor":false,"prefix":"","firstName":"Fayokun","middleName":"","lastName":"Ranti","suffix":""},{"id":453365999,"identity":"f77c2970-713c-40e5-b82f-1608cbe5403e","order_by":6,"name":"Pratima Murthy","email":"","orcid":"","institution":"National Institute of Mental Health and Neurosciences (NIMHANS)","correspondingAuthor":false,"prefix":"","firstName":"Pratima","middleName":"","lastName":"Murthy","suffix":""}],"badges":[],"createdAt":"2025-03-13 10:23:24","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6219017/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6219017/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41598-026-35417-5","type":"published","date":"2026-01-30T15:58:03+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":82270866,"identity":"642bc700-b772-4e87-96f4-6c1ed874bbfd","added_by":"auto","created_at":"2025-05-08 14:08:22","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":98061,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eChromatogram of Nicotine along with calibration curve\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Figure1.png","url":"https://assets-eu.researchsquare.com/files/rs-6219017/v1/57472f470528a2a0ea193e24.png"},{"id":82270221,"identity":"033722f7-193b-44b7-8b35-61a7302f76f1","added_by":"auto","created_at":"2025-05-08 14:00:22","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":49118,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eChromatogram showing Humectants\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-6219017/v1/8168cc39f696d16ba422e8b1.png"},{"id":82270230,"identity":"582f7a8e-99ef-4125-9eeb-be8d103fe5ff","added_by":"auto","created_at":"2025-05-08 14:00:23","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":101623,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eChromatogram, m/z, and Calibration of Benzo [A] Pyrenes\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Figure3.png","url":"https://assets-eu.researchsquare.com/files/rs-6219017/v1/4252203144b537d7bcd20d03.png"},{"id":82270868,"identity":"0a3d3f85-8789-4238-92cc-46d6a6b66e3a","added_by":"auto","created_at":"2025-05-08 14:08:23","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":77497,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eComparison of Nicotine, Moisture, CO, Tar, and TPM Cigarettes\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Figure4.png","url":"https://assets-eu.researchsquare.com/files/rs-6219017/v1/a48c487679362d2f4488f028.png"},{"id":82270867,"identity":"f36b503a-7df1-4cb5-b29a-0f774ede9a3c","added_by":"auto","created_at":"2025-05-08 14:08:23","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":75929,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eComparison of BAP and Humectants in Myanmar Cigarettes\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Figure5.png","url":"https://assets-eu.researchsquare.com/files/rs-6219017/v1/b06340fb59290580ee5f1f08.png"},{"id":82270236,"identity":"de6d896b-3ba1-4bde-9ec2-36461754e15b","added_by":"auto","created_at":"2025-05-08 14:00:23","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":89244,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eComparison of Moisture, TPM, CO, Tar, and Nicotine of Indian Cigarettes and Bidis\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Figure6.png","url":"https://assets-eu.researchsquare.com/files/rs-6219017/v1/5020ba1d763dd816b62d8656.png"},{"id":82270234,"identity":"4e4faf15-b95a-4051-99d9-a73dfeed1c7b","added_by":"auto","created_at":"2025-05-08 14:00:23","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":81875,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eComparison of BAP and Humectants \u0026nbsp;Cigarettes and Bidis\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Figure7.png","url":"https://assets-eu.researchsquare.com/files/rs-6219017/v1/53433dc6eeddc8283027a881.png"},{"id":82270878,"identity":"10569fd4-f598-4b97-b677-520146e44746","added_by":"auto","created_at":"2025-05-08 14:08:23","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":22500,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure 7: Humectant and BAP in Cigarettes and Beedis\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-6219017/v1/5fbfb39f45d0fea321278712.png"},{"id":101690410,"identity":"5504f07d-629b-4d72-8ec8-44b428aae5e1","added_by":"auto","created_at":"2026-02-02 16:01:07","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1382450,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6219017/v1/86b022bf-f522-4192-abe0-3667047c0cb1.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Analysis of Nicotine, Tar, CO, TPM, Moisture, BAP, and Humectants in Cigarettes and Beedis from India and Myanmar","fulltext":[{"header":"What this paper adds ","content":"\u003cp\u003e\u003cem\u003eCigarette smoking remains a global source of major health problems and is the leading cause of preventable deaths and diseases due to non-communicable diseases globally. This research aimed to examine the mainstream smoke deliveries from popular cigarettes and beedis from India and Myanmar. The smoke deliveries of Nicotine, CO, moisture, and tar in mainstream smoke from 11 cigarettes and 9 beedi brands were analyzed. Humectants (Propylene glycol, glycerol, and triethylene glycol) and benzo [A] pyrenes were also tested. The level of nicotine in the Indian brands was lower than in the Myanmar brands. However, the locally produced Indian beedis showed higher nicotine, moisture, CO, and tar levels compared to cigarettes.\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n"},{"header":"Introduction","content":"\u003cp\u003eTobacco use, including smoking, leads to over 8\u0026nbsp;million deaths annually, including over 7\u0026nbsp;million from direct tobacco use and roughly 1.2\u0026nbsp;million from exposure to passive smoking. \u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e. On average, smoking shortens life expectancy by about 10 years \u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e. Tobacco use is a major threat to public health, it kills around 4\u0026nbsp;million people every year in the WHO Southeast Asia Region \u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e. As per the fifth edition of the WHO global report on trends in the prevalence of tobacco use 2000\u0026ndash;2030, the SEARO is home to around 199\u0026nbsp;million tobacco smokers (20% of the global total) and 280\u0026nbsp;million smokeless tobacco users (77% of the global total). Tobacco is consumed in diverse forms, including smoked products, such as cigarettes and beedis (a cheaper hand-rolled form of smoked tobacco), and a variety of smokeless tobacco (SLT). Comprehensive awareness of tobacco\u0026rsquo;s harmful effects is far from desirable among users due to culture-related customs and misbeliefs about their health effects \u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eAll forms of tobacco are harmful, and there is no safe level of exposure to tobacco. Cigarette smoking is the most common form of tobacco use worldwide. Other smoked tobacco products include waterpipe, cigars, cigarillos, heated tobacco products (HTPs), roll-your-own tobacco, pipe tobacco, beedis, and kreteks. Around 80% of the 1.3\u0026nbsp;billion tobacco users worldwide live in low and middle-income countries \u003csup\u003e\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e, where the burden of tobacco-related illness and death is heaviest. Tobacco use contributes to poverty by diverting household spending from basic needs such as food, education, and shelter to buying tobacco products.\u003c/p\u003e \u003cp\u003eIndia is the second largest consumer and producer of tobacco in the world. The Global Adult Tobacco Survey 2017 revealed that 28.6% (around 266\u0026nbsp;million) of adults in India, aged 15 and above, currently use tobacco in some form. Of these, 10.7% (around 99\u0026nbsp;million) currently smoke tobacco and 21.4% (around 199\u0026nbsp;million) use smokeless tobacco \u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e. Tobacco products increase the risk of heart and lung diseases, as well as cancers and a variety of other adverse health outcomes \u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e. India's GDP is negatively impacted by tobacco use, and the direct medical expenses incurred in treating tobacco-related illnesses account for 5.3% of the nation's yearly spending on both public and private health care \u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e. It is estimated that the economic costs of tobacco-related diseases are ten times higher than the amount of money India receives from tobacco taxes \u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e. This drains the public health system and the economy, which no country can afford. In the Southeast Asian region, Myanmar reports the highest tobacco use, with over half of adults (54%) using some form of tobacco. STEPS 2014 further revealed that 26% of adults in Myanmar were tobacco smokers and 43% used smokeless tobacco \u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e,\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e. India and Myanmar can avert millions of preventable deaths as well as mitigate the damage of tobacco use on both society and the economy through the implementation of a comprehensive package of strong tobacco control policies outlined in the WHO Framework Convention on Tobacco Control (FCTC).\u003c/p\u003e \u003cp\u003eNicotine in tobacco is responsible for its addictive nature. Nicotine is a paradoxical substance because it has both stimulating and depressant effects. It affects the cardiovascular, skeletal, gastrointestinal, and peripheral nervous systems, among other systems \u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e. At the molecular levels, nicotine in tobacco mimics the neurotransmitter acetylcholine. Some of the effects of nicotine may be traced to its many receptors in cholinergic binding sites in the brain. Nicotine could be the main psychoactive component of tobacco due to its intricate pharmacological actions \u003csup\u003e\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/sup\u003e. Several additional essences are purposefully added to tobacco products to make them attractive and to lessen their harshness and potential for dependence \u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e. Many non-tobacco substances, such as humectants, fragrances, and flavors are added to attract and appeal to consumers. The American Lung Association has released data indicating that there are roughly 600 ingredients in one cigarette. Almost 7,000 chemicals are released when cigarettes burn, many of which are toxic. At least 69 of them are known to cause cancer \u003csup\u003e\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eTo ensure compliance with WHO FCTC Article 9 and effective regulatory controls, the contents and emissions of tobacco products must be monitored. The WHO Tobacco Free Initiative (TFI) established the tobacco testing laboratory network (TobLabNet) in 2005 as part of its global agenda. This is based on the WHO\u0026rsquo;s Study Group on Tobacco Product Regulation (TobReg) recommendation to build and strengthen tobacco product testing and research capacity by pursuing Articles 9 and 10 of the WHO Framework Convention on Tobacco Control (WHO FCTC). Under WHO TFI's leadership, TobLabNet develops and validates methods and standard operating procedures (SOPs) for testing the contents and emissions of tobacco products.\u003c/p\u003e \u003cp\u003eIn India, 29% of all adults use tobacco, while in Myanmar, over half of adults (54.4%) use some form of tobacco \u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e,\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e. A wide variety of smoked tobacco products are available in India and Myanmar. However, very little data on the emissions and addictiveness of smoked tobacco products is available from the region because of the required infrastructure and technical expertise involved in emission testing. This research aims to collect scientific information about the amount of nicotine, moisture, humectants, and benzo [A] pyrene in tobacco emissions using standard and validated methods and available laboratory support.\u003c/p\u003e"},{"header":"Material and Methods","content":"\u003cp\u003eThis study was conducted from June 2023 to May 2024 at the Tobacco Testing and Drug Toxicology Laboratory, Centre for Addiction Medicine (CAM), National Institute of Mental Health and Neurosciences (NIMHANS), Bengaluru, Karnataka. The state-of-the-art Drug Toxicology Laboratory is a world-class facility with high-end and sophisticated drug and tobacco testing instruments. The Tobacco Testing facility is a member of the World Health Organization (WHO) global network of Tobacco Testing Laboratories.\u003c/p\u003e \u003cp\u003eAll the chemicals used for analysis were of analytical grade. Nicotine, n-Heptadecane (Internal standard), Benzo[A]Pyrene and Humectants (Glycerol, Propylene Glycol, Triethylene Glycol), 1, 3- Butanediol (Internal standard), Flavors (Methyl Salicylate, Ethyl Salicylate, Eugenol, Di-phenyl ether, Cinnamaldehyde, Menthol, Coumarin, Camphor), 3\u0026prime;, 4\u0026prime;-Methylenedioxy acetophenone (MDA) (Internal standard) and Solvents (Cyclohexane, Propan-2-ol, Benzene, Methanol) procured through Sigma Aldrich, USA having purity of \u0026ge;\u0026thinsp;99.0%.\u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e\u003cb\u003e2.1 Patients and Public Involvement\u003c/b\u003e: Patients were not involved in this research.\u003c/h2\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Sample Collection and Storage:\u003c/h2\u003e \u003cp\u003eSamples were randomly selected from the cigarette sales points. Four cigarettes and nine Beedi brand samples were procured locally from wholesale and retail outlets in and around Bangalore, India. Seven cigarette brands sourced from Myanmar were obtained through WHO, SEARO, New Delhi, India. Reference cigarettes (3R4F, 1R5F, 2R4F) were obtained from the University of Kentucky. CORESTA Monitor (CM4 and CM6) reference cigarettes were received from CORESTA (Paris, France).\u003c/p\u003e \u003cp\u003eThe samples were transported in air-tight sealed packs to the Tobacco Testing Facility, NIMHANS, Bengaluru. The labeled samples were stored in sealed packs within 10 days of receipt at \u0026ndash; 20\u0026deg;C (Ultra Low deep freezer from Vestfrost) in their original packaging following the International Organization for Standardization (ISO, Geneva, Switzerland) guidance document ISO 3402:1999). Before analysis, samples were refrigerated for 24 h for comprehensive re-equilibration, followed by 2 h of equilibration to ambient conditions in the Environmental Chamber, Thermo Scientific 3900 Series \u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003e2.3 Ethics approval and consent to participate\u003c/h3\u003e\n\u003cp\u003eEthical approval was not required as this work did not involve human subjects.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e3.1 Instrumentation\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFor accurate testing of smoking behavior, it is crucial to replicate human smoking patterns using a smoke machine. This machine simulates human smoking by controlling the frequency and depth of inhalation, allowing researchers to analyze various factors such as chemical concentration, consistency, and total quantity.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe Cerulean SM450i is a 20-channel semi-automatic linear smoking machine designed to replicate human smoking behavior and complies with the International Organization for Standardization (ISO 3308) standards, ensuring reliable results and regulatory compliance. It can handle a diverse range of cigarettes/beedis without sacrificing efficiency or throughput. Beedi production in India is unregulated and lacks automation. \u0026nbsp;The tendu leaves used for wrapping are less combustible and nonporous, stronger puffs with greater volume and frequency may be required to ignite and maintain a consistent burn. The ISO 3308 standards may not be suitable for studying beedi emissions a relevant instrument and approach are necessary for accurate beedis analysis. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eISO 17175 was preferred for beedis emission analysis. The sampling procedure entails puffing for two seconds at 30-second intervals. The Cerulean beedi holder (Part: 99301) and latex beedi sleeve (Part: KU0358) were used in conjunction with a handheld vacuum pump to smoke beedis. The emissions were collected in bags, and the carbon monoxide (CO) percentage by volume was measured using a non-dispersive infrared (NDIR) analyzer. Total Particulate Matter (TPM) was calculated as per ISO 3308 by weighing the holders before and after the smoke run \u003csup\u003e9\u003c/sup\u003e. \u0026nbsp;The particulate matter collected on Cambridge filter pads (44 mm) was extracted as per TobLabNet SOP-10 for nicotine quantification. \u0026nbsp;Nicotine quantification was done on an Agilent 8890 Gas Chromatograph with a Flame ionization detector. The chromatographic separation was achieved on CP-WAX 51, column (CP7405) in isothermal mode at 170\u0026ordm; C shown in Figure 1.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eHumectants such as glycerol, propylene glycol, and triethylene glycol are added to tobacco products to facilitate the processing of the cured tobacco leaf, retain moisture, and increase shelf life. They dilute nicotine harshness and improve the sensory properties of tobacco products. Humectant quantification was done by TobLabNet SOP 6 using Agilent 8890 GC with FID. The humectants were separated on DB Wax (123-7032) fused silica column shown in Figure 2.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eBenzo[A]Pyrene belongs to the group of compounds known as polycyclic aromatic hydrocarbons (PAHs). PAHs are produced by heating or incomplete burning of organic material. They are not present in tobacco but are mostly produced as they burn. Exposure to BAP while smoking is one of the most important causes of cancer. Benzo[A]Pyrenes were quantified using TobLabNet SOP 5 on 5975 MSD clubbed with Agilent 7890 GC-MSD system. Chromatographic separation was achieved on the DB-5MS column (122-5532G). The mass spectra of ions produced by electro-spraying was identified by the National Institute of Standards and Technology (NIST) library and shown in Figure 3.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.2 Statistical Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSPSS Statistics for Windows, version 23.0. 2015 (SPSS Inc., Chicago, Ill., USA) was used to perform the statistical analysis of the data. A comparison between cigarettes and beedis was done through non-parametric analysis. Mann\u0026ndash;Whitney U-test was used for data with P \u0026lt; 0.05 was considered significant\u003cstrong\u003e.\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eThe comparisons of nicotine, CO, moisture, and tar between the cigarette samples received from Myanmar and India are summarised in Figure 4. The mean nicotine and CO in Myanmar cigarettes was 0.84 mg/cig (\u0026plusmn;0.17), and 13.35 mg/cig (\u0026plusmn;3.90) respectively. In Indian cigarettes, the mean nicotine was 0.66 mg/cig \u0026plusmn;0.08, and CO was 11.01 mg/cig \u0026plusmn;1.14. Although the nicotine and CO levels in Myanmar cigarettes were slightly higher than in Indian cigarettes, the difference was not statistically significant (Figure 4).\u003c/p\u003e\n\u003cp\u003eThe mean moisture content in Indian cigarettes was 4.44 \u0026plusmn;0.74, while for Myanmar cigarettes, it was 4.17 \u0026plusmn;1.93 but the difference was statistically insignificant. The mean Tar and TPM \u0026nbsp;in the Indian products were 10.34 \u0026plusmn; 2.03 mg/cig, and \u0026nbsp;15.44 \u0026plusmn; 2.13 mg/cig respectively, while the \u0026nbsp; Myanmar products had a mean \u0026nbsp;Tar 12.02\u0026plusmn; 4.05 and TPM \u0026nbsp;17.03 \u0026plusmn; 2.13 mg/cig however, the difference was not statistically significant (Figure 4).\u003c/p\u003e\n\u003cp\u003eThe mean Benzo[A]Pyrene in Indian cigarettes was 8.24 \u0026plusmn; 0.28, while the cigarettes from Myanmar cigarettes reported high BAP levels with a mean of 12.17\u0026plusmn; 2.24. A statistically significant (\u0026le;0.008) variation was observed in BAP levels (Figure 5). \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eHumectants are added to cigarettes to control and maintain the moisture content of the tobacco filler. The mean glycerol levels in Indian cigarettes were 10.27 mg/gm \u0026plusmn; 5.65, while the mean propylene glycol was 0.38 mg/gm \u0026plusmn; 0.30 and triethylene glycol was 0.03 mg/gm \u0026plusmn; 0.04. Myanmar cigarettes had mean glycerol levels of 10.02 mg/gm \u0026plusmn; 5.08, propylene glycol, 2.52 mg/gm \u0026plusmn; 1.32, and triethylene glycol 0.02 mg/gm \u0026plusmn; 0.06. On comparing all three humectants, we found there was no statistically significant variation in the glycerol and triethylene glycol, but propylene glycol was statistically significant (p \u0026le; 0.023) (Figure 5).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eBeedi had mean nicotine levels of 1.36 mg/beedi \u0026plusmn;0.56 and mean CO levels of 13.95 mg/beedi \u0026plusmn;2.43 (Figure 6). The moisture content of beedis varied from 1.32 to 28.4 mg/beedi, with a mean of 18.27 mg/beedi \u0026plusmn; 5.95. The mean Tar levels were 19.22 \u0026plusmn; 11.94 and TPM levels were 38.85 mg/beedi \u0026plusmn; 7.80. On comparing Indian Cigarettes with Beedis we report Nicotine, CO \u0026nbsp;(p \u0026le;0.02), moisture, and TPM (\u0026le;0.008 ) \u0026nbsp;were statistically significant. \u0026nbsp;When Beedis were compared with cigarettes from both countries we found that nicotine (p \u0026le;0.041), moisture (p \u0026le;0.001), TPM (p \u0026le;0.001), propylene glycol (p \u0026le;0.002), and glycerol \u0026nbsp;(p \u0026le;0.001) differ significantly.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eBeedis had mean glycerol levels 0.91mg/gm \u0026plusmn; 0.58, propylene glycol was 0.09mg/gm \u0026plusmn; 0.06, and triethylene glycol levels 0.04 mg/gm \u0026plusmn;0.03. On comparing beedi humectants with cigarettes, we found that values were statistically significant for glycerol (p \u0026le;0.009) only (Figure 7). BAP for cigarettes had a mean of 8.24\u0026plusmn;0.28 while for beedis it was 10.22 \u0026plusmn;1.07, the values were statistically significant (p \u0026le;0.008).\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;All cigarettes and beedis emissions were tested for flavors (Methyl Salicylate, Ethyl Salicylate, Eugenol, Eucalyptol, Di phenyl Ether, Cinnamaldehyde, Menthol, Coumarin, and Camphor) using Centre for Disease Control and Prevention (CDC) SOP, 60. Flavors were not detected in the samples received from both countries.\u0026nbsp;\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eCigarettes are expertly crafted, mass-produced items available worldwide. They are filled with processed tobacco and wrapped in paper. \u0026nbsp;Beedis are small, hand-rolled tobacco products made of sun-dried and processed tobacco wrapped in Tendu or Temburni leaves and secured with cotton thread\u0026nbsp;\u003csup\u003e9\u003c/sup\u003e. India has the world\u0026apos;s second-largest number of adult smokers. Beedi is the most commonly smoked tobacco product smoked by an estimated 71.8 million adults \u003csup\u003e2,4\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTobacco smoke is a complex mixture of over seven thousand volatile and semi-volatile compounds. Particulate matter (PM) generated due to tobacco smoking can be coarse (\u0026le;10 \u0026micro;m), fine (\u0026le;2.5 \u0026micro;m), or ultrafine (\u0026le;1 \u0026micro;m) \u003csup\u003e11\u003c/sup\u003e. The size of PM indicates both its ability to adsorb toxic organic compounds and the depth to which it can enter the respiratory system. The protonated form of nicotine gets deposited on the PM and gradually absorbed by the body. The other compounds of tobacco smoke are humectants, volatile aldehydes, nitrogen oxides, and polyacrylic aromatic hydrocarbons. Many harmful and potential carcinogens are present in mainstream smoke. If inhaled, high levels of carbon monoxide, especially from beedis can enter the bloodstream and bind with hemoglobin to form carboxy-hemoglobin. This results in tissue ischemia, which is a major cause of cardiovascular diseases, common among beedi smokers \u0026nbsp;\u003csup\u003e10\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe Government of India signed the Framework Convention on Tobacco Control (FCTC) (in 2004) and enacted the Cigarettes and Other Tobacco Products Act (COTPA) in 2003. The COTPA has not imposed any specific limitations on emissions or tobacco content. In its advisory note on the Global Nicotine Reduction Strategy, the WHO Study Group on Tobacco Product Regulation (TobReg) stated that the risk of dependence on cigarettes can be reduced by dropping their nicotine content to a very low level. \u0026nbsp;The nicotine content should therefore be as low as is technically feasible. \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAccording to WHO\u0026apos;s worldwide scan of national laws pertaining to maximum emission levels of tar, nicotine, and carbon dioxide (TNCO), as of December 31, 2022, eighty-three, nations had established maximum levels of nicotine (N) permitted per cigarette, with sixty of these nations (72%) allowing one milligram of nicotine per cigarette. Furthermore, sixty-seven countries permit ten milligrams of tar per cigarette, while eighty-five countries have set maximum levels of tar (T) per cigarette. Additionally, fifty-seven nations have set a carbon monoxide restriction of ten milligrams/ cigarette worldwide. In the Southeast Asian region, only Myanmar expressly forbids the exhibition of emission levels, At the same time Timor-Leste has established limits of ten milligrams for tar, one milligram for nicotine, and ten milligrams for carbon monoxide per cigarette (TNCO of 10/1/10 mg/cigarette). \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAll the cigarette brands tested had nicotine levels (0.77 mg/cigarette \u0026plusmn;0.16) within acceptable limits. For the beedis, the mean nicotine levels 1.36 \u0026nbsp;mg/beedi \u0026plusmn; 0.56 were above the maximum set limit (1.0mg). Ten milligrams of carbon monoxide per cig is the standard maximum permissible limit. Cigarette samples (n=6) received from Myanmar were found to exceed the common maximum allowable limit of 10mg carbon monoxide/cig. All beedis samples had CO levels above the set limit, the mean was 13.95 mg/beedi \u0026plusmn;2.43. Beedis smokers are potentially exposed to significantly higher concentrations of nicotine due to the greater puffing frequency. Research has shown that the non-porous nature and higher moisture content of tendu leaf in beedis led to higher levels of carbon monoxide and tar in its smoke compared to regular cigarette smoke. The presence of harmful and carcinogenic chemicals in mainstream beedi smoke could be harmful to human health.\u003csup\u003e11\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003eThe Myanmar cigarettes had Tar levels \u0026ge;12.02mg of Tar/cig \u0026plusmn;4.05, which was well above the highest limit (10.00mg/cig) set by countries. Indian beedis also had mean \u0026nbsp;Tar levels well above the accepted limit (19.22 mg /beedi\u0026plusmn;11.94 ). \u0026nbsp;Humectants including propylene glycol and glycerol were present in all the samples from both countries while trimethylene glycol was not detected in a few samples. \u0026nbsp;Indian cigarettes and beedis had propylene glycol levels \u0026le; 1.0 mg/gm, and glycerol in Indian brands was \u0026ge;10 mg/gm except for one make which had very low glycerol levels (1.99 mg/gm). Triertylene glycol in Indian cigarettes was not detected in all the samples, only two samples had it in the range of 0.04 - 0.09 mg/gm.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eYan Xizheg tested twenty-seven popular cigarette brands in the USA for humectants. \u0026nbsp;Only four brands showed no detectable amounts of the humectants, and the rest of the brands showed the presence of glycerol or 1,2-propylene glycol with concentrations ranging from 1.66 - 3.57% and from 0.23 to 1.35% for glycerol and 1,2-propylene glycol, respectively. In general, all the manufacturers use more glycerol than 1,2-propylene glycol as a humectant agent which may serve as a precursor for the formation of harmful carbonyl compounds\u0026nbsp;\u003csup\u003e12\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eBenzo(a)pyrene (BAP) is a marker of carcinogenic activity of polycyclic aromatic hydrocarbon (PAH). PAH is present in emission exhausts in small quantities of cigarette smoke, typically \u0026lt;10 ng/cigarette. In Indian brands of cigarettes, BAP was \u0026le; 10 ng/cig. In the cigarettes received from Myanmar only two brands had \u0026nbsp;BAP levels 9.95 and 9.72 ng/cig. In the rest of the samples, it was \u0026ge; 10.0 ng/cig with a mean of 10.22 ng/beedi\u0026plusmn;1.07 which may be harmful to the users depending upon the consistency of use. Information regarding BAP levels in the products offered by SEARO is limited. The analysis of mainstream smoke of cigarettes from Nigeria revealed that the BAP levels varied between 0 and 22.7 ng/cig. The most prevalent PAH in all of the products examined was naphthalene, which ranged from 210.7 to 460.34 ng/ \u0026nbsp;cig\u0026nbsp;\u003csup\u003e13\u003c/sup\u003e. \u0026nbsp;\u0026nbsp;\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eCigarettes and beedis present a significant challenge to the public\u0026rsquo;s health. Their availability and allure could override the health risks and negative perceptions attached to them. A quantitative analysis of emissions in mainstream smoke is necessary to assess the harmful effects of smoking. Current research was planned to quantitatively analyze Nicotine, CO, Moisture, Tar, TPM, humectants, and BAP, in the mainstream smoke of selected cigarette and beedi samples from India and Myanmar. Some popular brands of cigarettes (India and Myanmar) and beedis were analyzed using globally accepted SOPs. of ISO, CORESTA, and TobLabNet. \u0026nbsp; We report statistically significant differences in BAP (\u0026le;0.008) and Triethylene glycol (\u0026le;0.023) values, between Indian and Myanmar cigarettes. On comparing Indian cigarettes with beedis nicotine, CO (\u0026le;0.023) and glycerol (\u0026le;0.001) showed a statistically significant difference. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe contents and emissions were measured using globally approved protocols. This information can be further utilized to make policy decisions, educate the public about the dangers of smoking, and potentially incentivize tobacco companies to produce less harmful products by setting standards for emission levels of toxins.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eLimitation:\u003c/strong\u003e Analysis of tobacco-specific nitrosamines could not be planned in the absence of reference standards. \u0026nbsp;\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eB[A]P: Benzo[A]Pyrene\u003c/p\u003e\n\u003cp\u003eCAM: Centre for Addiction Medicine\u003c/p\u003e\n\u003cp\u003eCDC: Centre for Disease Control and Prevention \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eCFP: Cambridge Filter Pad\u003c/p\u003e\n\u003cp\u003eFID: Flame Ionisation Detector\u003c/p\u003e\n\u003cp\u003eGCMS: Gas Chromatography-Mass Spectrometry\u003c/p\u003e\n\u003cp\u003eISO: International Standard Organisation\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eNDIR: Non- dispersive infrared Analyser\u003c/p\u003e\n\u003cp\u003eNIMHANS: National Institute of Mental Health and Neurosciences\u003c/p\u003e\n\u003cp\u003eTFI: Tobacco Free Initiative\u003c/p\u003e\n\u003cp\u003eSOP: Standard Operating Procedure\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eWHO: World Health Organisation\u003c/p\u003e\n\u003cp\u003eSEARO: South East Asia Regional Office\u003c/p\u003e\n\u003cp\u003eTobLabNet: Tobacco Laboratory Network\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthors and Contributions:\u003c/strong\u003e Prof. Pratima Murthy and Dr. Jagdish Kaur conceptualized this idea. Dr. Priyamvada Sharma wrote the primary manuscript. Ranti F helped writing and interpretation of results. Dr. Arvind V Rinkoo helped in the final editing of the manuscript. Prof. P Marimutthu performed the statistical analysis. Ms. Amina Salam and Ms. Vijayahree Rao conducted all the experimental work at NIMHANS, Bangalore, India. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e: None declared\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u0026nbsp;\u003c/strong\u003eThis project was funded by WHO under the grant WHO Registration: 2023/1365732-0. PO: 2023166156\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData sharing statement:\u003c/strong\u003e The datasets used and/or analysed during the current study are available from the corresponding author upon reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDisclaimer:\u0026nbsp;\u003c/strong\u003eThe content is solely the responsibility of the authors and does not necessarily represent the views of the Institutes involved.\u0026nbsp;\u003c/p\u003e\n"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eKaur, J., Rinkoo, A. V. \u0026amp; Richardson, S. Update on numbers of tobacco-attributable deaths by country in the South-East Asia region: policy implications. \u003cem\u003eTob Control\u003c/em\u003e tc-2024-058599 (2024) doi:10.1136/tc-2024-058599.\u003c/li\u003e\n\u003cli\u003e(Global Burden of Disease [database. Washington, DC: Institute of Health Metrics; 2019. IHME, accessed 17 July 2023).\u003c/li\u003e\n\u003cli\u003eChen, D. T.-H. \u003cem\u003eet al.\u003c/em\u003e A longitudinal study of transitions between smoking and smokeless tobacco use from the ITC Bangladesh Surveys: implications for tobacco control in the Southeast Asia region. \u003cem\u003eThe Lancet Regional Health - Southeast Asia\u003c/em\u003e \u003cstrong\u003e14\u003c/strong\u003e, 100185 (2023).\u003c/li\u003e\n\u003cli\u003e(WHO Global Report on trends in prevalence of tobacco use 2000-2025, fourth edition. WHO, Geneva, 20215).\u003c/li\u003e\n\u003cli\u003eEconomic Costs of Diseases and Deaths Attributable to Tobacco Use in India, 2017\u0026ndash;2018.\u003c/li\u003e\n\u003cli\u003eMurray, J. B. Nicotine as a psychoactive drug. \u003cem\u003eJ Psychol\u003c/em\u003e \u003cstrong\u003e125\u003c/strong\u003e, 5\u0026ndash;25 (1991).\u003c/li\u003e\n\u003cli\u003e\u003cem\u003eAction on Smoking and Health. What\u0026rsquo;s in a Cigarette (2022). Https://Ash.Org.Uk/Resources/View/Whats-in-a-Cigarette\u003c/em\u003e.\u003c/li\u003e\n\u003cli\u003eSharma, P. \u003cem\u003eet al.\u003c/em\u003e Physical and chemical characterization of smokeless tobacco products in India. \u003cem\u003eSci Rep\u003c/em\u003e \u003cstrong\u003e13\u003c/strong\u003e, 8901 (2023).\u003c/li\u003e\n\u003cli\u003eOladipupo, O. A., Dutta, D. \u0026amp; Chong, N. S. Analysis of chemical constituents in mainstream bidi smoke. \u003cem\u003eBMC Chemistry\u003c/em\u003e \u003cstrong\u003e13\u003c/strong\u003e, 93 (2019).\u003c/li\u003e\n\u003cli\u003eGuaraldi, G. \u003cem\u003eet al.\u003c/em\u003e Lung and Heart Diseases Are Better Predicted by Pack-Years than by Smoking Status or Duration of Smoking Cessation in HIV Patients. \u003cem\u003ePLoS ONE\u003c/em\u003e \u003cstrong\u003e10\u003c/strong\u003e, e0143700 (2015).\u003c/li\u003e\n\u003cli\u003eOladipupo, O. A., Dutta, D. \u0026amp; Chong, N. S. Analysis of chemical constituents in mainstream bidi smoke. \u003cem\u003eBMC Chemistry\u003c/em\u003e \u003cstrong\u003e13\u003c/strong\u003e, 93 (2019).\u003c/li\u003e\n\u003cli\u003eXizheng, Y., Valent\u0026iacute;n-Blasini, L., Watson, C. \u0026amp; Cardenas, R. B. Determination of Humectants in Tobacco Filler by High Performance Chromatography/Single Quadrupole Mass Spectrometry. \u003cem\u003eBeitr\u0026auml;ge zur Tabakforschung International/Contributions to Tobacco Research\u003c/em\u003e \u003cstrong\u003e28\u003c/strong\u003e, 170\u0026ndash;178 (2018).\u003c/li\u003e\n\u003cli\u003eAdesina, O. A., Olowolafe, T. I. \u0026amp; Igbafe, A. Levels of polycyclic aromatic hydrocarbon from mainstream smoke of tobacco products and its risks assessment. \u003cem\u003eJournal of Hazardous Materials Advances\u003c/em\u003e \u003cstrong\u003e5\u003c/strong\u003e, 100053 (2022).\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Nicotine, Tar, Carbon Monoxide, TPM, Benzo[A]Pyrene, Moisture, Humectants, Flavors, cigarettes","lastPublishedDoi":"10.21203/rs.3.rs-6219017/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6219017/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eApproximately 2.3\u0026nbsp;million annual deaths in the eleven countries of the WHO Southeast Asia Region (SEAR) are linked to tobacco smoking. In 2020, smoking was responsible for 1.6\u0026nbsp;million lives lost in SEAR. The toxic substances found in the emissions of smoked tobacco products are inadequately researched. This study aims to evaluate and compare the smoke delivery potential of nicotine, tar, carbon monoxide, humectants, tobacco particulate matter (TPM), benzo[a]pyrene, and moisture between traditional beedis and commercial cigarettes. For quantifying emission ingredients from both beedis and cigarettes, globally accepted procedures from TobLabNet, the CDC, and CORESTA were utilized.\u003c/p\u003e \u003cp\u003eWhen comparing Indian and Myanmar cigarettes, we discovered that nicotine and carbon monoxide levels in Myanmar cigarettes were slightly higher than those in Indian cigarettes, though the difference was statistically insignificant. Moisture, tar, and TPM also exhibited statistically insignificant variations. The mean benzo[a]pyrene levels in Indian and Myanmar products were 8.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28 and 12.17\u0026thinsp;\u0026plusmn;\u0026thinsp;2.24, respectively, with a statistically significant difference (\u0026le;\u0026thinsp;0.008). Among humectants, only propylene glycol showed significant variation (p\u0026thinsp;\u0026le;\u0026thinsp;0.023).\u003c/p\u003e \u003cp\u003eThe comparison between Indian cigarettes and beedis revealed statistically significant differences in nicotine (p\u0026thinsp;\u0026le;\u0026thinsp;0.041), moisture (p\u0026thinsp;\u0026le;\u0026thinsp;0.001), TPM (p\u0026thinsp;\u0026le;\u0026thinsp;0.001), propylene glycol (p\u0026thinsp;\u0026le;\u0026thinsp;0.002), and glycerol (p\u0026thinsp;\u0026le;\u0026thinsp;0.001). The mean benzo[a]pyrene for cigarettes was 8.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28, while for beedis, it was 10.22\u0026thinsp;\u0026plusmn;\u0026thinsp;1.07, demonstrating a statistically significant difference (p\u0026thinsp;\u0026le;\u0026thinsp;0.008), along with glycerol (p\u0026thinsp;\u0026le;\u0026thinsp;0.009) among humectants. When comparing beedis to cigarettes from both countries, we found significant differences in nicotine (p\u0026thinsp;\u0026le;\u0026thinsp;0.041), moisture (p\u0026thinsp;\u0026le;\u0026thinsp;0.001), TPM (p\u0026thinsp;\u0026le;\u0026thinsp;0.001), propylene glycol (p\u0026thinsp;\u0026le;\u0026thinsp;0.002), and glycerol (p\u0026thinsp;\u0026le;\u0026thinsp;0.001). Flavors were not detected in the mainstream smoke of the cigarettes and beedis analyzed for this study.\u003c/p\u003e \u003cp\u003eThe findings of this study can be leveraged to enhance public health by identifying harmful chemicals that exceed established limits and potentially motivating manufacturers to produce less harmful products by conforming to toxin emission standards.\u003c/p\u003e","manuscriptTitle":"Analysis of Nicotine, Tar, CO, TPM, Moisture, BAP, and Humectants in Cigarettes and Beedis from India and Myanmar","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-05-08 14:00:18","doi":"10.21203/rs.3.rs-6219017/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-06-25T03:57:09+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-05-29T03:17:07+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"258063158521297893774984688437746193529","date":"2025-05-28T02:31:08+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"326841806788619557719750990730329340831","date":"2025-05-26T08:38:03+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-05-14T09:29:41+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"291721576600507539817979758335252061149","date":"2025-05-08T12:27:12+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-05-06T04:03:01+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-04-30T06:39:10+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-03-20T15:41:54+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-03-20T10:28:32+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2025-03-13T10:13:17+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"be284d14-2b6e-45d2-a2ca-357c67866e93","owner":[],"postedDate":"May 8th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":48213843,"name":"Health sciences/Health care"},{"id":48213844,"name":"Physical sciences/Chemistry"}],"tags":[],"updatedAt":"2026-02-02T16:00:01+00:00","versionOfRecord":{"articleIdentity":"rs-6219017","link":"https://doi.org/10.1038/s41598-026-35417-5","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2026-01-30 15:58:03","publishedOnDateReadable":"January 30th, 2026"},"versionCreatedAt":"2025-05-08 14:00:18","video":"","vorDoi":"10.1038/s41598-026-35417-5","vorDoiUrl":"https://doi.org/10.1038/s41598-026-35417-5","workflowStages":[]},"version":"v1","identity":"rs-6219017","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6219017","identity":"rs-6219017","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}