Pair your accounts.

Export articles to Mendeley

Get article recommendations from ACS based on references in your Mendeley library.

Pair your accounts.

Export articles to Mendeley

Get article recommendations from ACS based on references in your Mendeley library.

You’ve supercharged your research process with ACS and Mendeley!

Click to create an ACS ID

Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

Your Mendeley pairing has expired. Please reconnect
ACS Publications. Most Trusted. Most Cited. Most Read
My Activity

Figure 1Loading Img
ADDITION / CORRECTIONThis article has been corrected. View the notice.

Analysis of 23 Polycyclic Aromatic Hydrocarbons in Smokeless Tobacco by Gas Chromatography−Mass Spectrometry

View Author Information
Masonic Cancer Center, University of Minnesota, Mayo Mail Code 806, 420 Delaware Street SE, Minneapolis, Minnesota 55455
* To whom correspondence should be addressed. Tel: 612-624-4998. Fax: 612-626-5135. E-mail: [email protected]
Cite this: Chem. Res. Toxicol. 2010, 23, 1, 66–73
Publication Date (Web):October 27, 2009
Copyright © 2009 American Chemical Society

    Article Views





    Other access options


    Abstract Image

    Smokeless tobacco contains 28 known carcinogens and causes precancerous oral lesions and oral and pancreatic cancer. A recent study conducted by our research team identified eight different polycyclic aromatic hydrocarbons (PAHs) in U.S. moist snuff, encouraging further investigations of this group of toxicants and carcinogens in smokeless tobacco products. In this study, we developed a gas chromatography−mass spectrometry method that allows simultaneous analysis of 23 various PAHs in smokeless tobacco after a simple two-step extraction and purification procedure. The method produced coefficients of variation under 10% for most PAHs. The limits of quantitation for different PAHs varied between 0.3 and 11 ng/g tobacco, starting with a 300 mg sample. The recovery of the stable isotope-labeled internal standards averaged 87%. The method was applied to analysis of 23 moist snuff samples that included various flavors of the most popular U.S. moist snuff brands, as well as 17 samples representing the currently marketed brands of spit-free tobacco pouches, a relatively new type of smokeless tobacco. The sum of all detected PAHs in conventional moist snuff averaged 11.6 (±3.7) μg/g dry weight; 20% of this amount was comprised of carcinogenic PAHs. The levels of PAHs in new spit-free tobacco products were much lower than those in moist snuff; the sum of all detected PAHs averaged 1.3 (±0.28) μg/g dry weight. Our findings render PAHs one of the most prevalent groups of carcinogens in smokeless tobacco. Urgent measures are required from the U.S. tobacco industry to modify manufacturing processes so that the levels of these toxicants and carcinogens in U.S. moist snuff are greatly reduced.

    Read this article

    To access this article, please review the available access options below.

    Get instant access

    Purchase Access

    Read this article for 48 hours. Check out below using your ACS ID or as a guest.


    Access through Your Institution

    You may have access to this article through your institution.

    Your institution does not have access to this content. You can change your affiliated institution below.

    Cited By

    This article is cited by 65 publications.

    1. Francisco Gomez, Olalekan Ayo-Yusuf, Katrina Yershova, Vipin Jain, Aleksandra Alcheva, Dorothy K. Hatsukami, Mark Parascandola, Irina Stepanov. Heterogeneity of Harmful Constituent Profiles in Smokeless Tobacco Products from Five African Countries. Chemical Research in Toxicology 2023, 36 (12) , 1901-1911.
    2. Selvin H. Edwards, Matthew Hassink, Kenneth M. Taylor, An T. Vu. Variation of Benzo[a]pyrene, NNN, and NNK Levels in 16 Commercial Smokeless Tobacco Products. Chemical Research in Toxicology 2023, 36 (2) , 202-212.
    3. Clement N. Uguna, Colin E. Snape. Should IQOS Emissions Be Considered as Smoke and Harmful to Health? A Review of the Chemical Evidence. ACS Omega 2022, 7 (26) , 22111-22124.
    4. Karam El-Bayoumy, Kun-Ming Chen, Shang-Min Zhang, Yuan-Wan Sun, Shantu Amin, Gary Stoner, and Joseph B. Guttenplan . Carcinogenesis of the Oral Cavity: Environmental Causes and Potential Prevention by Black Raspberry. Chemical Research in Toxicology 2017, 30 (1) , 126-144.
    5. An T. Vu, Kenneth M. Taylor, Matthew R. Holman, Yan S. Ding, Bryan Hearn, and Clifford H. Watson . Polycyclic Aromatic Hydrocarbons in the Mainstream Smoke of Popular U.S. Cigarettes. Chemical Research in Toxicology 2015, 28 (8) , 1616-1626.
    6. Yan Zhong, Steven G. Carmella, J. Bradley Hochalter, Silvia Balbo, and Stephen S. Hecht. Analysis of r-7,t-8,9,c-10-Tetrahydroxy-7,8,9,10-tetrahydrobenzo[a]pyrene in Human Urine: A Biomarker for Directly Assessing Carcinogenic Polycyclic Aromatic Hydrocarbon Exposure Plus Metabolic Activation. Chemical Research in Toxicology 2011, 24 (1) , 73-80.
    7. Stephen S. Hecht, Steven G. Carmella, Peter W. Villalta and J. Bradley Hochalter. Analysis of Phenanthrene and Benzo[a]pyrene Tetraol Enantiomers in Human Urine: Relevance to the Bay Region Diol Epoxide Hypothesis of Benzo[a]pyrene Carcinogenesis and to Biomarker Studies. Chemical Research in Toxicology 2010, 23 (5) , 900-908.
    8. Yong Li, Tao Pang, Junli Shi, Xingyu Liu, Zhaoli Xu, Zhongbang Song, He Xie. Determination of tobacco-specific nitrosamines in tobacco and mainstream cigarette smoke using one-step clean-up coupled with liquid chromatography- tandem mass spectrometry. Journal of Chromatography A 2023, 1710 , 464436.
    9. Monica L. Vermillion Maier, Lisbeth K. Siddens, Jamie M. Pennington, Sandra L. Uesugi, Edwin M. Labut, Emily A. Vertel, Kim A. Anderson, Lane G. Tidwell, Susan C. Tilton, Ted J. Ognibene, Kenneth W. Turteltaub, Jordan N. Smith, David E. Williams. Impact of phenanthrene co-administration on the toxicokinetics of benzo[a]pyrene in humans. UPLC-accelerator mass spectrometry following oral microdosing. Chemico-Biological Interactions 2023, 382 , 110608.
    10. Chima Ugbor, Geoffrey Okoeguale, Uzochiamaka Nwobodo, Nneka Duhu, Kenneth Udeme, Solomon Ekoh, Emmanuel Odo, Gregory Aji. Tobacco Snuff-Induced Hepatic Alterations. Journal of Pharmacology and Toxicology 2023, 18 (3) , 120-131.
    11. Selvin H. Edwards, Matthew D. Hassink, Kenneth M. Taylor, An T. Vu. Quantitative measurement of harmful and potentially harmful constituents, pH, and moisture content in 16 commercial smokeless tobacco products. Regulatory Toxicology and Pharmacology 2022, 133 , 105199.
    12. Fatin Samara, Isra Arshad Alam, Yehya ElSayed. Midwakh: Assessment of Levels of Carcinogenic Polycyclic Aromatic Hydrocarbons and Nicotine in Dokha Tobacco Smoke. Journal of Analytical Toxicology 2022, 46 (3) , 295-302.
    13. Monica L. Vermillion Maier, Lisbeth K. Siddens, Jamie M. Pennington, Sandra L. Uesugi, Kim A. Anderson, Lane G. Tidwell, Susan C. Tilton, Ted J. Ognibene, Kenneth W. Turteltaub, Jordan N. Smith, David E. Williams. Benzo[a]pyrene (BaP) metabolites predominant in human plasma following escalating oral micro-dosing with [14C]-BaP. Environment International 2022, 159 , 107045.
    14. Jingjing Yu, Kejian Liu, Sheng Wang, Junwei Guo, Xiaoyu Wang, Li Ding, Junwei Zhao, Bing Wang, Junlan Cai, Li Chen, Shaofeng Liu, Wei Liu. Analytical determination of CML and CEL in oral use tobacco products by HPLC-MS/MS method. Microchemical Journal 2021, 168 , 106338.
    15. Do-Yeong Kim, Bo-Eun Lee, Han-Seung Shin. Determination of polycyclic aromatic hydrocarbons (PAHs) in smoking cessation aids by using high-performance liquid chromatography. Analytical Biochemistry 2021, 617 , 114119.
    16. Barbara Loken, Eugene Borgida, Tzushuo Wang, Molly K Madzelan, Allison L Williams, Dorothy Hatsukami, Irina Stepanov. Can the Public Be Educated About Constituents in Smokeless Tobacco? A Three-Wave Randomized Controlled Trial. Nicotine & Tobacco Research 2021, 23 (1) , 161-170.
    17. Chukwujindu M.A. Iwegbue, Kenneth O. Osijaye, Ufuoma A. Igbuku, Francis E. Egobueze, Godswill O. Tesi, Francisca I. Bassey, Bice S. Martincigh. Effect of the number of frying cycles on the composition, concentrations and risk of polycyclic aromatic hydrocarbons (PAHs) in vegetable oils and fried fish. Journal of Food Composition and Analysis 2020, 94 , 103633.
    18. Sushobhan Biswas, Hrishita Das, Ujjal Das, Aaveri Sengupta, Rakhi Dey Sharma, Subhas Chandra Biswas, Sanjit Dey. Smokeless tobacco induces toxicity and apoptosis in neuronal cells: a mechanistic evaluation. Free Radical Research 2020, 54 (7) , 477-496.
    19. Shamema Nasrin, Gang Chen, Christy J. W. Watson, Philip Lazarus, . Comparison of tobacco-specific nitrosamine levels in smokeless tobacco products: High levels in products from Bangladesh. PLOS ONE 2020, 15 (5) , e0233111.
    20. Serban C. Moldoveanu, Andrew Harrison. Extraction of Benzo[ a ]pyrene from Moist Snuff with Water or Artificial Saliva (Part 2). Contributions to Tobacco & Nicotine Research 2020, 29 (1) , 21-26.
    21. Yu-Ching Cheng, Carolyn M. Reyes-Guzman, Carol H. Christensen, Brian L. Rostron, Kathryn C. Edwards, Lanqing Wang, Jun Feng, Jeffery M. Jarrett, Cynthia D. Ward, Baoyun Xia, Heather L. Kimmel, Kevin Conway, Carmine Leggett, Kristie Taylor, Charlie Lawrence, Ray Niaura, Mark J. Travers, Andrew Hyland, Stephen S. Hecht, Dorothy K. Hatsukami, Maciej L. Goniewicz, Nicolette Borek, Benjamin C. Blount, Dana M. van Bemmel. Biomarkers of Exposure among Adult Smokeless Tobacco Users in the Population Assessment of Tobacco and Health Study (Wave 1, 2013–2014). Cancer Epidemiology, Biomarkers & Prevention 2020, 29 (3) , 659-667.
    22. Irina Stepanov, Dorothy K. Hatsukami. Chemical characterization of smokeless tobacco products and relevant exposures in users. 2020, 121-150.
    23. Jasmine Kaur, Arun Sharma, Amit Kumar, Deeksha Bhartiya, Dhirendra Narain Sinha, Suchitra Kumari, Ruchika Gupta, Ravi Mehrotra, Harpreet Singh. SLTChemDB: A database of chemical compounds present in Smokeless tobacco products. Scientific Reports 2019, 9 (1)
    24. Kevin G. McAdam, Harriet Kimpton, Arif Faizi, Andrew Porter, Brad Rodu. The composition of contemporary American and Swedish smokeless tobacco products. BMC Chemistry 2019, 13 (1)
    25. Adama Kamissoko, Vincent Carré, Sébastien Schramm, Frédéric Aubriet. Study of the mainstream cigarette smoke aerosols by Fourier transform ion cyclotron resonance mass spectrometry coupled to laser/desorption and electrospray ionization – Additional insights on the heteroaromatic components. Rapid Communications in Mass Spectrometry 2019, 33 (S1) , 95-108.
    26. Serban C. Moldoveanu, Jerry W. Marshall, Thomas H. Poole. Extraction from Moist Snuff with Artificial Saliva of Benzo[ a ]pyrene and Other Polycyclic Aromatic Hydrocarbons. Beiträge zur Tabakforschung International/Contributions to Tobacco Research 2019, 28 (5) , 214-223.
    27. Filippo Zanetti, Alain Sewer, Bjoern Titz, Walter K. Schlage, Anita R. Iskandar, Athanasios Kondylis, Patrice Leroy, Emmanuel Guedj, Keyur Trivedi, Ashraf Elamin, Florian Martin, Stefan Frentzel, Nikolai V. Ivanov, Manuel C. Peitsch, Julia Hoeng. Assessment of a 72-hour repeated exposure to Swedish snus extract and total particulate matter from 3R4F cigarette smoke on gingival organotypic cultures. Food and Chemical Toxicology 2019, 125 , 252-270.
    28. Yuesong Wang, Lee-Yang Wong, Lei Meng, Erin N. Pittman, Debra A. Trinidad, Kendra L. Hubbard, Alisha Etheredge, Arseima Y. Del Valle-Pinero, Rachel Zamoiski, Dana M. van Bemmel, Nicolette Borek, Vyomesh Patel, Heather L. Kimmel, Kevin P. Conway, Charles Lawrence, Kathryn C. Edwards, Andrew Hyland, Maciej L. Goniewicz, Dorothy Hatsukami, Stephen S. Hecht, Antonia M. Calafat. Urinary concentrations of monohydroxylated polycyclic aromatic hydrocarbons in adults from the U.S. Population Assessment of Tobacco and Health (PATH) Study Wave 1 (2013–2014). Environment International 2019, 123 , 201-208.
    29. Ravi Prakash Yadav, Souvik Ghatak, Payel Chakraborty, Freda Lalrohlui, Ravi Kannan, Rajeev Kumar, Jeremy L. Pautu, John Zomingthanga, Saia Chenkual, Rajendra Muthukumaran, Nachimuthu Senthil Kumar. Lifestyle chemical carcinogens associated with mutations in cell cycle regulatory genes increases the susceptibility to gastric cancer risk. Environmental Science and Pollution Research 2018, 25 (31) , 31691-31704.
    30. Deeksha Bhartiya, Amit Kumar, Jasmine Kaur, Suchitra Kumari, Amitesh Kumar Sharma, Dhirendra N. Sinha, Harpreet Singh, Ravi Mehrotra. In-silico study of toxicokinetics and disease association of chemicals present in smokeless tobacco products. Regulatory Toxicology and Pharmacology 2018, 95 , 8-16.
    31. Laura Romito. Perioperative Management of the Tobacco User. 2018, 345-358.
    32. Ziyan Fan, Zhonghao Li, Shanshan Liu, Fei Yang, Zhaoyang Bian, Ying Wang, Gangling Tang, Qinxiao Zhao, Huimin Deng, Shili Liu. Rapid fluorescence immunoassay of benzo[ a ]pyrene in mainstream cigarette smoke based on a dual-functional antibody–DNA conjugate. RSC Advances 2018, 8 (52) , 29562-29569.
    33. Robert Nilsson. Use of rodent data for cancer risk assessment of smokeless tobacco in the regulatory context. Regulatory Toxicology and Pharmacology 2017, 88 , 338-348.
    34. Karl A. Wagner, Chorng B. Huang, Matt S. Melvin, Regina Ballentine, Naren K. Meruva, Jason W. Flora. Gas Chromatography-Mass Spectrometry Method to Quantify Benzo[a]Pyrene in Tobacco Products. Journal of Chromatographic Science 2017, 55 (7) , 677-682.
    35. Bin Lai, Manuel R. Plan, Nils J.H. Averesch, Shiqin Yu, Frauke Kracke, Nicolas Lekieffre, Sarah Bydder, Mark P. Hodson, Gal Winter, Jens O. Krömer. Quantitative analysis of aromatics for synthetic biology using liquid chromatography. Biotechnology Journal 2017, 12 (1)
    36. Sushobhan Biswas, Krishnendu Manna, Ujjal Das, Amitava Khan, Anirban Pradhan, Aaveri Sengupta, Surajit Bose, Saurabh Ghosh, Sanjit Dey. Smokeless tobacco consumption impedes metabolic, cellular, apoptotic and systemic stress pattern: A study on Government employees in Kolkata, India. Scientific Reports 2016, 5 (1)
    37. Katrina Yershova, Jian‐Min Yuan, Renwei Wang, Liza Valentin, Clifford Watson, Yu‐Tang Gao, Stephen S. Hecht, Irina Stepanov. Tobacco‐specific N ‐nitrosamines and polycyclic aromatic hydrocarbons in cigarettes smoked by the participants of the Shanghai Cohort Study. International Journal of Cancer 2016, 139 (6) , 1261-1269.
    38. Imabong A. Effiong, Francisca I. Bassey, Chukwujindu M. A. Iwegbue, Okon D. Ekpa, Sunday A. Williams, Fehintola C. Oguntunde, Vincent N. Osabor, Bice S. Martincigh. Polycyclic aromatic hydrocarbons in three commercially available fish species from the Bonny and Cross River estuaries in the Niger Delta, Nigeria. Environmental Monitoring and Assessment 2016, 188 (9)
    39. Arnab Ganguli, Amlan Das, Debasish Nag, Surela Bhattacharya, Gopal Chakrabarti. Potential role of autophagy in smokeless tobacco extract-induced cytotoxicity and in morin-induced protection in oral epithelial cells. Food and Chemical Toxicology 2016, 90 , 160-170.
    40. Min-Ae Song, Catalin Marian, Theodore M. Brasky, Sarah Reisinger, Mirjana Djordjevic, Peter G. Shields. Chemical and toxicological characteristics of conventional and low-TSNA moist snuff tobacco products. Toxicology Letters 2016, 245 , 68-77.
    41. Orish Ebere Orisakwe, Zelinjo Nkeiruka Igweze, Kenneth Obinna Okolo, Nnaemeka Arinze Udowelle. Human health hazards of poly aromatic hydrocarbons in Nigerian smokeless tobacco. Toxicology Reports 2015, 2 , 1019-1023.
    42. M. Carradus, K. G. McAdam, J. D. H. van Heemst, C. H. A. Goss, C. Wright. A validated method to measure benzo[a]pyrene concentrations in tobacco by high-performance liquid chromatography-fluorescence detection. Analytical Methods 2015, 7 (4) , 1590-1599.
    43. Ingeborg Lund, Karl Lund. How Has the Availability of Snus Influenced Cigarette Smoking in Norway?. International Journal of Environmental Research and Public Health 2014, 11 (11) , 11705-11717.
    44. Sébastien Schramm, Vincent Carré, Jean-Luc Scheffler, Frédéric Aubriet. Active and passive smoking – New insights on the molecular composition of different cigarette smoke aerosols by LDI–FTICRMS. Atmospheric Environment 2014, 92 , 411-420.
    45. Michael Schivo, Mark V. Avdalovic, Susan Murin. Non-Cigarette Tobacco and the Lung. Clinical Reviews in Allergy & Immunology 2014, 46 (1) , 34-53.
    46. Kevin G McAdam, Arif Faizi, Harriet Kimpton, Andrew Porter, Brad Rodu. Polycyclic aromatic hydrocarbons in US and Swedish smokeless tobacco products. Chemistry Central Journal 2013, 7 (1)
    47. S. Balbo, S. James-Yi, C. S. Johnson, M. G. O'Sullivan, I. Stepanov, M. Wang, D. Bandyopadhyay, F. Kassie, S. Carmella, P. Upadhyaya, S. S. Hecht. (S)-N'-Nitrosonornicotine, a constituent of smokeless tobacco, is a powerful oral cavity carcinogen in rats. Carcinogenesis 2013, 34 (9) , 2178-2183.
    48. Amlan Das, Abhijit Bhattacharya, Subhendu Chakrabarty, Arnab Ganguli, Gopal Chakrabarti, . Smokeless Tobacco Extract (STE)-Induced Toxicity in Mammalian Cells is Mediated by the Disruption of Cellular Microtubule Network: A Key Mechanism of Cytotoxicity. PLoS ONE 2013, 8 (7) , e68224.
    49. Tameka S. Lawler, Stephen B. Stanfill, Liqin Zhang, David L. Ashley, Clifford H. Watson. Chemical characterization of domestic oral tobacco products: Total nicotine, pH, unprotonated nicotine and tobacco-specific N-nitrosamines. Food and Chemical Toxicology 2013, 57 , 380-386.
    50. Lucy Popova, Pamela M. Ling. Alternative Tobacco Product Use and Smoking Cessation: A National Study. American Journal of Public Health 2013, 103 (5) , 923-930.
    51. K. E. Lund, A. McNeill. Patterns of Dual Use of Snus and Cigarettes in a Mature Snus Market. Nicotine & Tobacco Research 2013, 15 (3) , 678-684.
    52. . Bibliography. 2013, 1475-1712.
    53. M.F. Borgerding, J.A. Bodnar, G.M. Curtin, J.E. Swauger. The chemical composition of smokeless tobacco: A survey of products sold in the United States in 2006 and 2007. Regulatory Toxicology and Pharmacology 2012, 64 (3) , 367-387.
    54. Neal L. Benowitz, Caroline C. Renner, Anne P. Lanier, Rachel F. Tyndale, Dorothy K. Hatsukami, Bruce Lindgren, Irina Stepanov, Clifford H. Watson, Connie S. Sosnoff, Peyton Jacob. Exposure to Nicotine and Carcinogens among Southwestern Alaskan Native Cigarette Smokers and Smokeless Tobacco Users. Cancer Epidemiology, Biomarkers & Prevention 2012, 21 (6) , 934-942.
    55. Sanjit Mukherjee, Aneek Das Bhowmik, Paromita Roychoudhury, Kanchan Mukhopadhyay, Jay Gopal Ray, Keya Chaudhuri. Association of XRCC1, XRCC3, and NAT2 polymorphisms with the risk of oral submucous fibrosis among eastern Indian population. Journal of Oral Pathology & Medicine 2012, 41 (4) , 292-302.
    56. Robert Nilsson. The molecular basis for induction of human cancers by tobacco specific nitrosamines. Regulatory Toxicology and Pharmacology 2011, 60 (2) , 268-280.
    57. Ziad S. Naufal, Kristin M. Marano, Steven J. Kathman, Cody L. Wilson. Differential exposure biomarker levels among cigarette smokers and smokeless tobacco consumers in the National Health and Nutrition Examination Survey 1999–2008. Biomarkers 2011, 16 (3) , 222-235.
    58. M. A. Macha, A. Matta, S. S. Chauhan, K. W. M. Siu, R. Ralhan. Guggulsterone (GS) inhibits smokeless tobacco and nicotine-induced NF- B and STAT3 pathways in head and neck cancer cells. Carcinogenesis 2011, 32 (3) , 368-380.
    59. Muzafar A. Macha, Ajay Matta, Shyam Singh Chauhan, K. W. Michael Siu, Ranju Ralhan, . Guggulsterone Targets Smokeless Tobacco Induced PI3K/Akt Pathway in Head and Neck Cancer Cells. PLoS ONE 2011, 6 (2) , e14728.
    60. Olalekan A Ayo-Yusuf, Greg N Connolly. Applying toxicological risk assessment principles to constituents of smokeless tobacco products: implications for product regulation. Tobacco Control 2011, 20 (1) , 53-57.
    61. K. Frost-Pineda, S. Appleton, M. Fisher, K. Fox, C. L. Gaworski. Does Dual Use Jeopardize the Potential Role of Smokeless Tobacco in Harm Reduction?. Nicotine & Tobacco Research 2010, 12 (11) , 1055-1067.
    62. . Current Awareness in Drug Testing and Analysis. Drug Testing and Analysis 2010, 507-522.
    63. Andrew R. Joyce, William Hawkins, Marc W. Fariss, Tapas K. Sengupta. Role of plasma membrane disruption in reference moist smokeless tobacco-induced cell death. Toxicology Letters 2010, 198 (2) , 191-199.
    64. . Current literature in mass spectrometry. Journal of Mass Spectrometry 2010, 951-962.
    65. Coral E. Gartner, Wayne D. Hall. Smokeless tobacco products: Unrecognised regulatory and policy opportunities. Drug and Alcohol Review 2010, 29 (2) , 119-120.