nitrosamines analysis in tobacco cigarettesRecently, a story in Science Daily reported that the chemical (S)-N’-nitrosonornicotin [(S)-NNN] used in smokeless tobacco products is a strong oral carcinogen! This was the first study that identified a specific chemical present is these tobacco products as causing oral cancer in animals. Also, very interestingly at the same time, the state of Madhya Pradesh in India was the first state in that country to ban the sale of gutka (a mild stimulant consisting of crushed areca or betel nut that is chewed to get a mild buzz) containing tobacco or nicotine! This ban went into effect April 1, 2012. The state authorities say they have taken this decision after a much higher report of mouth cancers.

In the US, the US Dept of Health and Human Services has long reported the dangers of tobacco and tobacco smoke as in this comprehensive report from 2006:The Health Consequences of Involuntary Exposure to Tobacco Smoke: A Report of the Surgeon General (downloadable PDF). In addition, in 2009, the United States passed the H.R. 1256 (111th): Family Smoking Prevention and Tobacco Control Act, which allows the US FDA to lower the amount of nicotine in tobacco products, ban candy flavorings that appeal to children, and block misleading labels such as Low Tar and Light. However, the law does not permit the FDA to ban nicotine or tobacco outright, only regulate what goes into tobacco products, make public the ingredients in tobacco products, and prohibit marketing campaigns geared towards children.

As a part of the implementation of Act H.R. 1256, the US FDA’s Center for Tobacco Products (CTP) released an initial list of 93 Harmful and Potentially Harmful Constituents (HPHCs) chemicals or chemical compounds in tobacco and tobacco products; and, recently, the CTP released a shortened list of 20 chemicals that must be reported on by tobacco companies by the end of 2012.

Here we present, Application Note 242, Robust and Fast Analysis of Tobacco-Specific Nitrosamines by LC-MS/MS, (downloadable PDF) presents the analysis of tobacco-specific nitrosamines (TSNA) which 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. The application note describes a robust and fast LC-MS/MS method using a rapid-separation liquid chromatography analytical column (Thermo Scientific Acclaim Acclaim RSLC column) for the analysis of four TSNAs in tobacco cigarettes after a simple sample preparation of ammonium acetate liquid extraction and filtration.

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The chromatographic separation of the four TSNAs was achieved within 3.5 min and these analytes were detected with great sensitivity and selectivity by tandem mass spectrometry using Selected Reaction Monitoring (SRM) detection. This method has been evaluated with respect to linearity, detection limits, precision, and accuracy. The ruggedness of the methodology was proven with more than 1000 replicate injections.

Next in this series, we will cover the analysis of environmental nicotine exposure by hydrophilic interaction liquid chromatography-tandem mass spectrometry, nicotine bioanalysis in urine, and an article on the analysis nitrosamines in water using automated solid-phase extraction & GC-MS!

Let us know if this application note was helpful to you in your work! We look forward to hearing from you.