Trace Analysis of Mutagenic Heterocyclic Aromatic Amines in Cigarette Smoke Condensate and its Base Fractions via Silylation-GC-MS

Open access

Abstract

Among the more than 5000 chemicals reported in cigarette smoke condensate (CSC), heterocyclic aromatic amines (HAAs) are considered to be a contributor to observed biological activity. HAAs are non-volatile and are reported at ppb levels in CSC. A new method for HAA analysis at the trace level is reported here. N, O-Bis(trimethylsilyl) trifluoroacetamide (BSTFA) containing 1% trimethylchlorosilane was employed to derivatize amino groups by heating the reagent containing a sample of CSC at 80 °C for 30 min followed by analysis employing gas chromatography-mass spectroscopy (GC-MS) in the selected-ion-monitoring (SIM) mode. This derivatization method afforded symmetrical peak shapes on a ZB-50 stationary phase and achieved instrumental limits of quantification (LOQ) at 10:1 S/N from -1 ng/mL for AαC to120 ng/mL for Glu-P-1. The chemical identity of each derivative was confirmed by comparison of retention time and mass spectra of standards. The latter were characterized by the following ions: M·+ or [M-1]+, [M-15]+, and m/z 73 (i.e., trimethylsilyl). CSC and its base sub-fractions were studied using the GC-MS method. Ten HAAs were screened and five were quantified in cigarette smoke condensate, while 2-5 HAAs were quantified in each of three base sub-fractions. Values obtained with the new procedure agree well with values reported in the literature and with results obtained from a commercial laboratory via a different analytical method. The potential contribution of each HAA to the overall mutagenic activity observed for CSC and its base fractions is discussed. When considered together, HAAs account for only a small portion (-7.8%) of the observed mutagenicity of the CSC.

1. Alaejos, M.S., J.H. Auala, V. Gonzalez, and A.M. Afonso: Analytical Methods Applied to the Determination of Heterocyclic Aromatic Amines in Foods; J. Chromatogr. B 862 (2008) 15–42.

2. International Agency for Research on Cancer (IARC): Some Naturally Occurring Substances: Food Items and Constituents, Heterocyclic Aromatic Amines and Mycotoxins, IARC Monographs of the Evaluation of Carcinogenic Risk to Humans; IARC, Lyon, France, IARC Monograph 56 (1993) 165–244.

3. Turesky, R.J., J. Taylor, L. Schnackenberg, J.P. Freeman, and R.D. Holland: Quantitation of Carcinogenic Heterocyclic Aromatic Amines and Detection of Novel Heterocyclic Aromatic Amines in Cooked Meats and Grill Scrapings by HPLC/ESI-ESI; J. Agric. Food Chem. 53 (2005) 3248–3258.

4. Food and Drug Administration (FDA): Harmful and Potentially Harmful Constituents in Tobacco Products and Tobacco Smoke, Established List; Federal Register 77(64) (2012) 20034–20037, available at: http://www.gpo.gov/fdsys/pkg/FR-2012-04-03/html/2 012-7727.htm (accessed 19th March 2013).

5. Poindexter, E.H. and R.D. Carpenter: The Isolation of Harman and Norharman from Tobacco and Cigarette Smoke; Phytochemistry 1 (1962) 215–212.

6. Hada, N., Y. Totsuka, T. Enya, K. Tsurumaki, M. Nakazawa, N. Kawahara, Y. Murakami, Y. Yoko-yama, T. Sugimura, and K. Wakabayashi: Structures of Mutagens Produced by the Co-Mutagen Norharman with o- and m-Toluidine Isomers; Mutat. Res. 493 (2001) 115-126.

7. Levitt, R.C., C. Legraverend, D.W. Nebert, and O. Pelkonen: Effects of Harman and Norharman on the Mutagenicity and Binding to DNA of Benzo[a]pyrene Metabolites in vitro and on Aryl Hydrocarbon Hydrolase Induction; Cell Cult. 79 (1977) 1167-1175.

8. Matsumoto, T., D. Yoshida, and H. Tomita: Determination of Mutagens, Amino-a-Carbolines in Grill Foods and Cigarette Smoke Condensate; Cancer Lett. 12 (1981) 105-110.

9. Patrianakos, C. and D. Hoffmann: Chemical Studies on Tobacco Smoke LXIV. On the Analysis of Aromatic Amines in Cigarette Smoke; J. Anal. Toxicol. 3 (1979) 150-154.

10. Yamashita, M., K. Wakabayashi, M. Nagao, S. Sato, Z. Yamaizum, M. Takashi, N. Kinac, I. Tomita, and T. Sugimura: Detection of 2-Amino-3-methyl-3H-imidazo[4,5-f]quinoline in Cigarette Smoke Condensate; Jpn J. Cancer Res. 77 (1986) 419-422.

11. Manabe, S., K. Tohyama, O. Wada, and T. Aramaki: Detection of a Carcinogen 2-Amino-1-methyl-6-phenyl-1 H-imidazo[4,5-b]pyridine (PHIP) in Cigarette Smoke Condensate; Carcinogenesis 12 (1991) 1945-1947.

12. Manabe, S. and O. Wada: Carcinogenic Tryptophan Pyrolysis Products in Cigarette Smoke Condensate and Cigarette Smoke-Polluted Indoor Air; Environ. Pollut. 64 (1990) 121-132.

13. Kataoka, H., K. Kijima, and G. Maruo: Determination of Mutagenic Heterocyclic Amines in Combustion Smoke Samples; Bull. Environ. Contam. Toxicol. 60 (1998) 60-76.

14. Kataoka, H. and K. Kijima: Analysis of Heterocyclic Amines as their N-Dimethylaminomethylene Derivatives by Gas Chromatography with Nitrogen-Selective Detection; J. Chromatogr. A 767 (1997) 187-194.

15. Sasaki, T.A., J.M. Wilkins, J.B. Forehand, and S.C. Moldoveanu: Analysis of Heterocyclic Amines in Mainstream Smoke Using a New GC-MS Technique; Anal. Lett. 34 (2001) 1749-1761.

16. Smith, C.J., G.L. Dooly, and S.C. Moldoveanu: New Technique Using Solid-Phase Extraction for the Analysis of Aromatic Amines in Mainstream Cigarette Smoke; J. Chromatogr. A 991 (2003) 99-107.

17. Smith, C.J., X. Qian, Q. Zha, and S.C. Moldoveanu: Analysis of a- and ß- Carbolines in Mainstream Smoke of Reference Cigarettes by Gas Chromatography-Mass Spectrometry; J. Chromatogr. A 1046 (2004) 211-216.

18. Zhang, L., D.L. Ashley, and C.H. Watson: Quantitative Analysis of Six Heterocyclic Aromatic Amines in Mainstream Smoke Condensate Using Isotope Dilution Liquid Chromatography-Electrospray Ionization Tandem Mass Spectrometry; Nicotine Tob. Res. 13 (2011) 120-126.

19. Pillsbury, H.C., CC. Bright, K.J. O`Connor, and F.W. Irish: Tar and Nicotine in Cigarette Smoke; J. AOAC 52 (1969) 458-462.

20. Meckley, D., J.R. Hayes, K.R. Van Kampen, A.T. Mosberg, and J.W. Swauger: A Responsive, Sensitive, and Reproducible Dermal Tumor Promotion Assay for the Comparative Evaluation of Cigarette Smoke Condensates; Regul. Toxicol. Pharmacol. 39 (2004) 135-149.

21. Iwase, H., Y. Takeuchi, and A. Murai: Gas Chromatography-Mass Spectrometry of Trimethylsilyl Derivatives of Amines; Chem. Pharm. Bull. 27 (1997) 1009-1014.

22. Narasimhachari, N., J. Spaide, and B. Heller: Gas-Liquid Chromatographic and Mass Spectrometric Studies on Trimethylsilyl Derivatives of N-methyl and N, N-dimethyltryptamines; J. Chromatogr. Sci. 9 (1971) 502-505.

23. Bao, M., J. Wu, W. Rickert, A. Masters, and P. Joza: Analysis of Heterocyclic Aromatic Amines in Mainstream Cigarette Smoke Condensate by Solid Phase Extraction and Ultra-Performance Liquid Chromatography Tandem Mass Spectrometry; Presentation at the 2012 CORESTA Congress in Sapporo Japan, Smoke Science/Product Technology Groups, abstr. SS18; 2012; available at: http:// www.coresta.org/Meetings/Past_Abstracts/ Sapporo2012-SmokeTech.pdf (accessed February 2013).

24. Maron, D.M. and B.N Ames: Revised Methods for the Salmonella Mutagenicity Test; Mutat. Res. 113 (1983) 173-215.

25. Yahagi, T., M. Degawa, Y. Seino, J. Matsushima, M. Nagao, T. Sugimura, and Y. Hashimoto: Mutagenicity of Carcinogenic Azo Dyes and their Derivatives; Cancer Lett. 1 (1975) 91-97.

26. Baty, J.D., D.A. Price-Evans, H.M. Gilles, and J. Greaves: Gas Chromatography-Mass Spectrometry of Trimethylsilyl Derivatives of Amines; Chem. Pharm. Bull. 27 (1997) 1009-1014.

27. Kataoka, H.: Methods for the Determination of Mutagenic Heterocyclic Amines and Their Applications in Environmental Analysis; J. Chromatogr. A 774 (1997) 121-142.

28. McLafferty, F.W. and F. Turecek: Interpretation of Mass Spectra; 4th Edition, University Science Books, Sausalito, CA, USA, 1993.

29. Sugimura, T.: Past, Present and Future of Mutagens in Cooked Foods; Environ. Health Persp. 67 (1986) 5-10.

30. Smith, C, V. Payne, DJ. Doolittle, A.K. Debath, T. Lawlor, and C. Hansch: Mutagenic Activity of a Series of Synthetic and Naturally Occurring Heterocyclic Amines in Salmonella; Mutat. Res. 279 (1992) 61-73.

31. Lee, CK., J.A. Munoz, C. Fulp, K. Chang, J.C. Rogers, M.F. Borgerding, and D.J. Doolittle: Inhibitory Activity of Cigarette-Smoke Condensate on the Mutagenicity of Heterocyclic Amines; Mutat. Res. 322 (1994) 21-32.

32. Roemer, E., H. Schramke, H. Weiler, A. Buettner, S. Kausche, S. Weber, A. Berges, M. Stueber, M. Muench, E. Trelles-Sticken, J. Pype, K. Kohlgrueber H. Voelkel, and S. Wittke: Mainstream Smoke Chemistry and in vitro and in vivo Toxicity of the Reference Cigarettes 3R4F and 2R4F; Beitr. Tabakforsch. Int. 25 (2012) 316-335.

Journal Information


CiteScore 2017: 0.63

SCImago Journal Rank (SJR) 2017: 0.309
Source Normalized Impact per Paper (SNIP) 2017: 0.403

Metrics

All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 329 321 42
PDF Downloads 257 256 36