A CORESTA Recommended Method (CRM 75) has been developed and published, applicable to the quantification of tobacco-specific nitrosamines (TSNAs), namely, Nnitrosonornicotine (NNN), N-nitrosoanabasine (NAB), Nnitrosoanatabine (NAT) and 4-(N-nitrosomethylamino)-1-(3-pyridyl)-1-butanone (NNK) in cigarette mainstream smoke. The method involves smoke collection on a Cambridge filter pad under both ISO 3308 and the intense conditions adopted by Health Canada. An internal standard solution is added to the smoke collected on the pad and, after extraction, an aliquot is separated and quantitatively analysed by liquid chromatographytandem mass spectrometry (LC-MS/MS).
CRM 63 involving gas chromatography coupled with a thermal energy analyser (GC-TEA) was previously developed by the CORESTA Special Analytes Group that had been set up to develop recommended methods on smoke components. However, by 2009 most laboratories had moved to similar LC-MS/MS methods for TSNA analysis and so this technique was chosen as the basis of a new CRM and to complement CRM 63. Initial joint experiments, specific experiments by single laboratories and ongoing discussions identified methodological aspects that needed to be ‘standardised’ before moving to a CRM.
A joint experiment by 15 laboratories was carried out in 2010-2011 that investigated and identified important methodological features that needed to be controlled or clarified. CRM 75 was produced through a final collaborative experiment involving 20 laboratories from 12 countries using both linear and rotary smoking machines. Some notes are included in the CRM to inform other laboratories that might wish to adopt the method, concerning aspects that need to be well controlled to provide data as robust as possible and to provide similar repeatability and reproducibility data.
Statistical evaluations were made according to ISO 5725 guidelines and are included. Under ISO smoking, the levels of reproducibility (R) expressed as a percentage of the mean of TSNA yields across laboratories are much greater than the levels found for “tar”, nicotine and carbon monoxide and given in the relevant ISO standards. The R value was expressed as a percentage of the mean yield amonglaboratories and across all of the studied products. Under
ISO smoking R% values ranged from 25-60% for NNN; from 31-85% for NNK; from 47-58% for NAT and 40-99% for NAB. These levels are generally in line with those determined previously for TSNAs in CRM 63 and for other smoke analytes studied by the Special Analytes Group.
Under ‘intense’ smoking, R% values ranged from 30-88% for NNN; from 37-79% for NNK; from 47-83% for NAT and 42-111% for NAB. A plot of R against mean yields suggests that the ‘intense’ regime gives similar or slightly worse reproducibility than the ISO regime in spite of the higher yields generated.
1. Health Canada: Tobacco Reporting Regulations (Ttr) (2000) and its Amendments (2005); available at http://www.hc-sc.gc.ca/hc-ps/tobac-tabac/legislation/ reg/indust/index-eng.php (accessed November 2012).
2. World Health Organization: The Scientific Basis of Tobacco Product Regulation; Second Report of a WHO Study Group (TobReg); WHO Technical Report Series 951 (2008) ISBN 978 92 4 120951 9.
3. Adams, J.D., K.D. Brunnemann, and D. Hoffmann: Chemical Studies on Tobacco Smoke. LXXV. Rapid Method for the Analysis of Tobacco Specific N-nitros-amines by Gas-liquid Chromatography with A Thermal Energy Analyser; J. Chromatogr. 256 (1983) 347–351.
4. Begutter, H., H. Klus, and I. Ultsch: Kapillargas-chromatographische Bestimmung flüchtiger und tabak-spezifischer N-Nitrosamine mittels des Thermo-Energy-Analyzers [Capillary Gas Chromatography Analysis of Volatile and Tobacco-specific N-nitros-amines by Means of Thermo-energy Analyzers]; J. Chromatogr. 321 (1985) 475–479.
5. Fischer, S. and B. Spiegelhalder: Improved Method for the Determination of Tobacco-specific Nitrosamines (Tsna) in Tobacco Smoke; Beitr. Tabakforsch. Int. 14 (1989) 145–153.
6. Health Canada: Determination of Nitrosamines in Mainstream Tobacco Smoke; 1999, available at http://www.hc-sc.gc.ca/hc-ps/tobac-tabac/legislation/ reg/indust/method/_main-principal/nitrosamines-eng.php (accessed November 2012).
7. CORESTA: CORESTA Recommended Method No 63; Determination of tobacco specific nitrosamines in cigarette mainstream smoke - GC-TEA method; 2005, available at http://www.coresta.org/Recommended_ Methods/CRM_63.pdf (accessed November 2012).
8. Wagner, K.A., N.H. Finkel, J.E. Fossett, and I.G. Gillman: 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 (2005) 1001–1006.
9. Wu, J., P. Joza, M. Sharifi, W.S. Rickert, and J.H. Lauterbach: 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 (2008) 1341–1345.
10. Xiong, W., H. Hou, X. Jiang, G. Tang, and Q. Hu: Simultaneous Determination of Four Tobacco-specific N-nitrosamines in Mainstream Smoke for Chinese Virginia Cigarettes by Liquid Chromatographytandem Mass Spectrometry and Validation under ISO and “Canadian Intense” Machine Smoking Regimes; Anal. Chim. Acta 674 (2010) 71–78.
11. International Organisation for Standardisation (ISO): International Standard ISO 3308:2000. Routine Analytical Cigarette-Smoking Machine. Definitions and Standard Conditions; 4th Edition, Geneva, Switzerland, 2000.
12. Health Canada: Determination of “Tar”, Nicotine and Carbon Monoxide in Mainstream Tobacco Smoke; 1999, available at http://www.hc-sc.gc.ca/hc-ps/tobac-tabac/legislation/reg/indust/method/_main-principal/nicotine-eng.php (accessed November 2012)
13. CORESTA: CORESTA Recommended Method No 75: Determination of Tobacco Specific Nitrosamines in Mainstream Cigarette Smoke by LC-MS/MS; June 2012, available at http://www.coresta.org/ Recommended_Methods/CRM_75.pdf (accessed November 2012).
14. International Organisation for Standardisation (ISO): International Standard ISO 5725-1:1994. Accuracy (Trueness and Precision) of Measurement Methods and Results - Part 1: General Principles and Definitions. ISO, Geneva, Switzerland, 1994.
15. International Organisation for Standardisation (ISO): International Standard ISO 5725-2:1994. Accuracy (Trueness and Precision) of Measurement Methods and Results - Part 2: Basic Method for the Determination of Repeatability and Reproducibility of a Standard Measurement Method. ISO, Geneva, Switzerland, 1994.
16. International Organisation for Standardisation (ISO): International Standard ISO 3402:1999. Tobacco and Tobacco Products - Atmosphere for Conditioning and Testing; ISO, Geneva, Switzerland, 1999.
17. International Organisation for Standardisation (ISO): International Standard ISO 4387:2000. Cigarettes -Determination of Total and Nicotine Free Dry Particulate Matter Using A Routine Analytical Smoking Machine; ISO, Geneva, Switzerland, 2000.
18. CORESTA: CORESTA Recommended Method No 70; Determination of Selected Volatile Organic Compounds in the Mainstream Smoke of Cigarettes - Gas Chro-matography-Mass Spectrometry Method, June 2010, available at http://www.coresta.org/Recommended_ Methods/CRM_70.pdf (accessed October 2012).
19. Saint-Jalm, Y., M.F. Borgerding, S.G. Chapman, and W.T. Morgan: Alternative smoking regimes Task Force. Final Report, 2006, available at http://www.coresta. org/Reports/Alternative-Smoking-Regimes-TF_Final Report_July2006.pdf (accessed October 2012).
20. Heller, W.D., H.J. Eberhardt, M. Czechowicz, D. Mariner, J. Sarabia, S.W. Purkis, and M. Meger: Explanation of Outlier Analysis - Remarks Based on the Evaluation of the WG10 Study. CORESTA Joint Meeting of the Smoke Science and Product Technology Study Groups, Graz, Austria, 2011, Paper ST 06.
21. CORESTA: CORESTA Recommended Method No 58. Determination of Benzo[a]pyrene in Cigarette Main-stream Smoke - Gas Chromatography-Mass Spectro-metry Method; 2004, available at http://www.coresta.org/Recommended_Methods/CRM_58.pdf (accessed October 2012).