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

Abstract

An improved liquid chromatography tandem mass spectrometry (LC-MS/MS) method has been developed for the determination of tobacco specific nitrosamines (TSNA). It utilizes four stable isotope-labeled internal standards instead of two as reported by others. A separate internal standard for each analyte is required to minimize sample matrix effects on each analyte, which can lead to poor analyte recoveries and decreases in method accuracy and precision if only one or two of the internal standards are used, especially for complex sample matrixes and when no sample cleanup steps are performed as in this study. In addition, two ion-transition pairs (instead of one) are used for each analyte for the confirmation and quantification, further enhancing the method's accuracy and robustness. These improvements have led to a new LC-MS/MS method that is faster, more sensitive, and selective than the traditional methods and more accurate and robust than the published LC-MS/MS methods. The linear range of the method was from 0.2 to 250 ng/mL with a limit of detection of each TSNA varied from 0.027 to 0.049 ng/mL. Good correlations between the results obtained by the new method and the traditional method were observed for the samples studied.

Rapid and sensitive gas chromatography–ion-trap tandem mass spectrometry method for the determination of tobacco-specific N-nitrosamines in secondhand smoke

“…In all cases, MDLs and MQLs were significantly below the levels determined in the SHS samples (see below). Furthermore, compared to other methods which involve a preconcentration step and clean-up procedure, MDLs for this method are much lower than for the GC/TEA method, are similar to those found using LC/MS/MS [15][16][17] and are better than those reported by Zhou et al [19] using GC-IT-MS/MS. Lower MDLs may be attained by using an accelerated solvent extraction (ASE) with lower solvent volumes, or employing a preconcentration step using rotary evaporator or solid phase cartridges (SPE).…”

“…However, the GC-TEA method has two main disadvantages: (1) it cannot differentiate the co-eluted nitroso-compounds although it is nitroso-specific, and (2) extensive sample preparation, including pre-concentration and clean-up using liquidliquid (L-L) extraction and/or solid phase extraction (SPE), is necessary due to the limited sensitivity of the method. Recently, LC-MS/MS methods have been developed for TSNA analysis, which provide greater sensitivity and selectivity than the GC-TEA method [15][16][17][18]. However, sample matrix effects, can lead to poor analyte recoveries and decreased accuracy and precision.…”

“…Sample clean up using L-L or SPE [15,18] can reduce July 13, 2009 5 these effects, but this increases analysis time and limits sample throughput. Another method uses stable isotopically-labeled standards [15][16][17]. Although this approach is effective, it significantly increases the cost of the analysis, making it impractical for routine use.…”

An UPLC‐MS3 Method for Rapid Separation and Determination of Four Tobacco‐specific Nitrosamines in Mainstream Cigarette Smoke

“…Although this technique can provide reasonable sensitivity and is relatively specific for nitrosamine compounds, it has difficulty in differentiating the coeluting nitroso compounds. 7 In recent years several HPLC-MS 2 methods, [13][14][15] which provide higher sensitivity and selectivity over the GC-TEA method, have been applied for the quantitative determination of TSNAs with simplified sample preparation. Despite high sensitivity and selectivity can be achieved with this technique, however, in some cases, the presence of complex matrix still lead to interferences in determination process.…”

Investigations into the pituri plant: nicotine content, nicotine conversion to nornicotine, nicotine release and cytotoxicity of Australian native Nicotiana spp.

“…Until recently the majority of these reports used GC coupled with thermal energy analyser (TEA) (Carmella et al, 2000, Stepanov et al, 2002 or mass spectrometry (MS) (Song and Ashley, 1999, Zhou et al, 2007, Sleiman et al, 2009), or HPLC with various detectors such as UV (Hecht et al, 1975), or TEA (Hoffmann et al, 1979). It is now more common to see LC-MS/MS being used extensively for quantitative analysis of TSNAs (Kim and Shin, 2013, Wagner et al, 2005, Wu et al, 2008, Ding et al, 2008, Xiong et al, 2010, Jansson et al, 2003 due to its high sensitivity.…”