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Clifford Watson, Ph.D. Centers for Disease Control and Prevention National Centers for Environmental Health Division of

Analytical methodologies for measuring harmful/potentially harmful chemicals in tobacco products/tobacco smoke . Clifford Watson, Ph.D. Centers for Disease Control and Prevention National Centers for Environmental Health Division of Laboratory Sciences June 8-9, 2010 Gaithersburg, Maryland.

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Clifford Watson, Ph.D. Centers for Disease Control and Prevention National Centers for Environmental Health Division of

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  1. Analytical methodologies for measuring harmful/potentially harmful chemicals in tobacco products/tobacco smoke Clifford Watson, Ph.D. Centers for Disease Control and Prevention National Centers for Environmental Health Division of Laboratory Sciences June 8-9, 2010 Gaithersburg, Maryland The findings and conclusions in this presentation have not been formally disseminated by the Centers for Disease Control and Prevention and should not be construed to represent any agency determination or policy.

  2. Objectives • Review general sources for tobacco products/tobacco smoke analytical methods • Terms, abbreviations, general overview of common analytical procedures • Review some commonly used methods for specific chemicals in tobacco/smoke • Multiple methods: Assessing equivalency, criteria to review relative strengths and weaknesses • Summary

  3. Sources for established tobacco products/tobacco smoke methods • International Organization for Standardization (ISO) • Cooperation Centre for Scientific Research Relative to Tobacco (CORESTA) • Health Canada • Commercial Labs (Labstat, Arista, Filtrona Scientific Services) • Centers for Disease Control and Prevention (CDC) • Tobacco Industry • Phillip Morris: Counts et al. (Reg. Tox and Pharm 39 (2004) 111-134) • British American Tobacco • Benchmark Study for Massachusetts

  4. Terms and abbreviations Method A standardized analytical laboratory procedure used to determine the amount or concentration of a specific chemical or group of chemicals Analyte A specific chemical (e.g., benzene) or mixture (e.g., tar) that is determined in an analytical laboratory procedure

  5. Example methods for specific chemicals in tobacco products/tobacco smoke Not an exhaustive listing Often multiple methods are available Not an endorsement Inclusion strictly illustrative

  6. General analytical procedure outline (common abbreviations) Generation (to be discussed at next meeting) Separation Gas chromatography (GC) High performance liquid chromatography (HPLC) Ion chromatography (IC) Detection Mass spectrometry (MS) Tandem mass spectrometry (MS/MS) Thermal energy analyzer (TEA) Thermal conductivity (TC) Flame ionization (FID) Conductivity (CD) Infrared Spectrometry (IR) Ultraviolet/Visible Spectroscopy (UV) Inductively Coupled Plasma (ICP) Atomic Absorption Spectroscopy (AAS)

  7. Example methods for specific analytes a) Total particulate matter; b) Tar is defined as TPM less the nicotine and water content; c) Carbon monoxide; d) Non-dispersive infrared detection 1) Health Canada T-115 2) ISO 10315:1991; 3) ISO 10362-1:1991; 4) Stanfill 2009; 5) ISO 4387:1991; 6) ISO 8454:1995

  8. Example methods, cont. 1) Health Canada T-116; 2) Arista Labs; 3) Dong et al. 2000; 4) Polzin et al. 2007

  9. Example methods, cont. 1) Health Canada T-104; 2) Polzin et al. 2007

  10. Example methods, cont. 1) Health Canada T-114; 2) Vaughan et al. 2008

  11. Example methods, cont. NNK: 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone; NNN: N’-nitrosonornicotine NAB: N’-nitrosoanabasine; NAT: N’-nitrosoanatabine 1) Hoffmann et al. 1979; 2) Truker et al. 1991; 3) Wu W et al. 2003; 4) Wagner, et al. 2005; 5) Wu J, et al. 2008.

  12. Example methods, cont. 1) Health Canada T-209; 2) Rhoades et al. 1987; 3) Health Canada T-206; 4) Pappas et al. 2006

  13. Example methods, cont. 1) Health Canada, T-102; 2) Health Canada, T-112

  14. Example methods, cont. 1) Health Canada, T-301, 1999; 2) Wu (2003)

  15. Example methods, cont. • Health Canada T-101; 2) Health Canada T-110; 3) Health Canada T-313; 4) Wilson et al. 2008

  16. Example methods, cont. 1) Health Canada T-304; 2) Health Canada T-103; 3) Ding et al. (2005); 4) Health CanadaT-107 5) Health Canada T-105; 6) ) Stanfill and Ashley (2000); 7) Health Canada T-308; 8) Health Canada T-312; 9) Health Canada T-311; 10) Celebucki et al (2005)

  17. Multiple methods Often different methods can provide comparable or equivalent results Standard criteria to establish equivalency exist: Representative sample set for comparison Analytical determination Statistical comparison of results

  18. Some considerations for selecting specific analytical methods Applicability Selectivity/Specificity Universal or chemical specific detection Analytical merit Quantification range, Linearity, Recovery, Robustness, Accuracy, Precision, Reproducibility,…

  19. Summary Analytical methods exist to quantify chemical levels for analytes on example list In many cases multiple analytical methods are available for measuring specific analytes Numerous factors guide decisions for using a specific method when multiple methods are available Means exist to compare analytical data from different methods to ensure comparability

  20. Clarifying Questions?

  21. Useful Web links for quick reference on tobacco/smoke method compilations • ISO www.iso.org • CORESTA www.coresta.org • Health Canada www.hc-sc.gc.ca • British American Tobacco www.bat-science.com • Labstat www.labstat.com/ • Arista Labs www.aristalabs.com 21

  22. References Arista Labs, http://www.aristalabs.com/customtesting_mainstream.htm Celebucki CC, et al. (2005) “Characterization of measured menthol in 48 U.S. cigarette sub-brands, ” Nicotine & Tobacco Research 7(4) 523–531. Ding YS, et al. (2005), “Determination of 14 Polycyclic Aromatic Hydrocarbons in Mainstream Smoke from Domestic Cigarettes,” Env. Sci. Tech., 39(2), 471-478. Dong JZ, et al. (2000), “A simple GC-MS technique for the analysis of vapor phase mainstream cigarette smoke. J. Microcolumn Separations 12, 142-148. Health Canada (1999) Determination of Ammonia in Mainstream Tobacco Smoke, Method T-101 Health Canada (1999) determination of Ni, Pb, Cd, Cr, As and Se in mainstream tobacco smoke, Method T-109 Health Canada (1999) Determination of 1- and 2- Aminonaphthalene and 3- and 4- Aminobiphenyl in Mainstream Tobacco Smoke, Method T-102 Health Canada (1999) Determination of Benzo[A]pyrene in Mainstream Tobacco Smoke, Official Method T-103 Health Canada (1999) Determination of Selected Carbonyls in Mainstream Tobacco Smoke, Method T-104 Health Canada (1999) Determination of Eugenol in Mainstream Tobacco Smoke, T-105 Health Canada (1999) Determination of Hydrogen Cyanide in Mainstream Tobacco Smoke, Method T-107 Health Canada (1999) Determination of Oxides of Nitrogen in Mainstream Tobacco Smoke, Method T-110 Health Canada (1999) Determination of Pyridine, Quinoline and Styrene in Mainstream Tobacco Smoke, Method T-112 Health Canada (1999) Determination of Phenolic Compounds in Mainstream Tobacco Smoke, Method T-114 Health Canada (1999) Determination of 1,3-Butadiene, Isoprene, Acrylonitrile, Benzene, and Toluene in Mainstream Tobacco Smoke, Method T-116 22

  23. References, cont. Health Canada (1999) Determination of Tar, Water, Nicotine and Carbon Monoxide in Mainstream Smoke, Method T-119 Health Canada (1999) Determination of mercury in sidestream tobacco smoke, Method T-206 Health Canada (1999) Determination of oxides of nitrogen in sidestream tobacco smoke, Method T-208 Health Canada (1999) Determination of Alkaloids in Whole Tobacco, Method T-301 Health Canada (1999) Determination of Humectants in Whole Tobacco, Method T-304 Health Canada (1999) Determination of Nitrate from Whole Tobacco, Method T-308 Health Canada (1999) Determination of Sodium Propionate in Whole Tobacco, Method T-312 Health Canada (1999) Determination of Triacetin in Whole Tobacco, Method T-311 Health Canada (1999) Determination of Sodium Propionate in Whole Tobacco, Method T-312 Health Canada (1999) Determination of Sorbic Acid in Whole Tobacco, Method T-313 Hoffmann et al. (1979), Assessment of Tobacco-specific N-Nitrosamines in Tobacco Products” Cancer Research, 39, 2505-2509 ISO 4387:1991, Determination of the total particulate matter present in the smoke. ISO 10362-1:1991, Water content of the total particulate. ISO 10315:1991, Nicotine content of the total particulate. ISO 8454:1995, Measurement of carbon monoxide present in the vapour phase of smoke Pappas RS, et al. (2006) Cadmium, lead, and thallium in mainstream tobacco smoke particulate. Food and Chemical Toxicology 44(5): 714-723. 23

  24. References, cont. Polzin et al. (2007), Analysis of volatile organic compounds in mainstream cigarette smoke. Env. Sci. Tech. 41, 1297-1303. Rhoades CB, et al.(1997) Mainstream smoke collection by electrostatic precipitation for acid dissolution in a microwave digestion system prior to trace metal determination. Journal of AOAC International 80(6):1320-1331. Stanfill SB, et al. (2000), “Quantification of flavor-related alkenylbenzenes in tobacco smoke particulate by selected ion monitoring gas chromatography mass spectrometry,” J. Arig. Food Chem., 48(4). 1298-1306. Truker et al. (1991), “N-Nitroso compounds in cigarette tobacco and their occurrence in mainstream tobacco smoke,” Carcinogenesis, 12(2), 257-261. Vaughan C, et al. (2008). Automated determination of seven phenolic compounds in mainstream tobacco smoke. Nicotine Tob Res. 10(7):1261-8. Wagner, et al. (2005), “Development of a Quantitative Method for the Analysis of Tobacco-Specific Nitrosamines in Mainstream Cigarette Smoke Using Isotope Dilution Liquid Chromatography/Electrospray Ionization Tandem Mass Spectrometry,” Anal. Chem., 77, 1001-1006. Wilson CL, et al. (2008), “Assessment of dioxin and dioxin-like compounds in mainstream smoke from selected US brands and reference cigarettes,” Food and Chem. Tox. 46, 1721-1733 Wu J, et al. (2008), “Quantitative Method for the Analysis of Tobacco Specific Nitrosamines in Cigarette Tobacco and Mainstream Cigarette Smoke by use of Isotope Dilution Liquid Chromatography Tandem Mass Spectrometry,” Anal. Chem, 80, 1341-1345 Wu W, et al.(2003) “Simultaneous Determination of Five Tobacco Specific Nitrosamines in Mainstream Cigarette Smoke by Isotope Dilution Liquid chromatography/Electrospray Ionization Tandem Mass Spectrometry,” Anal. Chem., 75(18), 4827 4832. 24

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