Fast Analysis of Tobacco Nitrosamines in Cigarettes by LC–MS–MS

Leo (Jinyuan) Wang and William C. Schnute, Dionex Corporation, Sunnyvale, California, USA.

Tobacco specific nitrosamines (TSNA) are a group of carcinogens found only in tobacco products and are formed from nicotine and related alkaloids during the production and processing of tobacco and tobacco products.¹ As a result of their carcinogenic properties, efforts have been made to reduce TSNA levels in tobacco products. The project goal was to demonstrate a high throughput and sensitive method to monitor TSNA levels.

Conventional methods for TSNA analysis are based on gas chromatography with thermo energy analyser (GC–TEA)^2,3 or high performance liquid chromatography (HPLC) with various detection techniques, such as UV and mass spectrometry (MS).^4–6 This application note describes a robust and fast LC–MS–MS method for the determination of four TSNAs in tobacco cigarettes.

Mass spectrometric conditions

See Figure 1.

Method Performance

Figure 1 shows the four TSNAs retained and completely resolved within 3.5 min, with the minimum of the retention factors (K'NNN) being greater than four and minimum resolution (RsNAT) greater than two. Demonstrated retention factors ensure the chromatographic separation of analytes of interest from early eluted compounds, which minimizes the possibility of ionization suppression and eliminates cross contamination of SRM transitions between main analytes. The use of specific MS–MS detection allows the simplification of sample preparation and the greater sensitivity ensures quantification confidence at low levels.

Figure 1

Because the analytical column was running under the very harsh conditions of high temperature and high pH mobile phase, method ruggedness was evaluated by repeated injections of standard solutions and tobacco extracts and was well maintained after more than 1000 injections.

Conclusion

A simple, sensitive and specific method for the determination of tobacco specific nitrosamines is demonstrated and has been applied to the analysis of TSNAs in different cigarettes.

References

1. S.S. Hecht and D. Hoffmann, Carcinogenesis, 9, 875–884 (1998).

2. J.D. Adams, K.D. Brunnemann and D. Hoffmann, J. Chromatog. A, 256, 347–351 (1983).

3. I.S. Krull et al., J. Chromatog. A, 260, 347–362 (1983).

4. W. Wu, D.L. Ashley and C.H. Watson, Anal. Chem., 75, 4827–4832 (2003).

5. K.A. Wagner et al., Anal. Chem., 77, 1001–1006 (2005).

6. J. Wu et al., Anal. Chem., 80, 1341–1345 (2008).

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