The purified samples were redissolved in CH3CN and analyzed by gas chromatography (GC) interfaced with a Thermal Energy Analyzer (Orion Research, Beverly, MA). Moisture content and pH were measured as previously described (Stepanov et al., 2008). Briefly, moisture content was measured via the difference www.selleckchem.com/products/ganetespib-sta-9090.html in weight of a tobacco sample before and after its drying for 3 hr at 99 ��C, while pH was measured in aqueous tobacco extracts. Nicotine and Unprotonated Nicotine Nicotine was measured as described elsewhere (Stepanov, Hecht, Ramakrishnan, & Gupta, 2005). Tobacco was extracted with MeOH containing KOH, and an aliquot of the extract was mixed with [CD3]nicotine internal standard.
The samples were transferred to GC-microinsert vials and analyzed by GC�Cmass spectrometry�Cselected ion monitoring using a model 6890 GC equipped with an autosampler and interfaced with a model 5973 mass selective detector (Agilent Technologies, Palo Alto, CA) as previously described (Stepanov et al., 2005). The amount of unprotonated nicotine was calculated using the Henderson�CHasselbalch equation based on the measured nicotine, pH, and pKa of 8.02 (Richter & Spierto, 2003). Statistical Analysis The statistical analysis was restricted to products for which three samples were obtained from different vendors in multiple locations around the country. Analysis of variance was conducted to compare the geometric means of the products. When the overall F test was significant, t tests were conducted to make the pairwise comparisons of interest. All p values were adjusted using the method of Bonferroni.
The level of significance was set to .05. Results A total of 117 samples were received as a part of New Product Watch: 71 samples of Marlboro Snus (Rich, Mild, Spearmint, and Peppermint), 36 samples of Camel Snus (Mellow, Frost, Robust, and Winterchill), 4 samples of Camel Orbs (Mellow and Fresh flavors), 3 samples of Camel Sticks (Mellow flavor), and 3 samples of Camel Strips (Fresh flavor). Marlboro Snus Rich, Mild, Spearmint and Peppermint, as well as Camel Snus Mellow and Frost were obtained from all six regions as planned. Samples from Mid-Atlantic/Appalachia were not included in the statistical analysis because they were procured from the same store. Only one sample of Camel Snus Robust and one sample of Camel Snus Winterchill were obtained from Paris, AR.
Camel dissolvables were found only by observers in Indiana. Camel Orbs were purchased in two locations (Spenser and Indianapolis), and Camel Sticks and Strips were purchased in three locations (Angola, Spenser, and Indianapolis). Thus, sample sizes for Camel Snus Robust and Winterchill and for Camel dissolvables were insufficient to make any statistical comparisons. TSNA and nicotine AV-951 were not detected in negative control samples. Analysis of NNN and NNK in Copenhagen Snuff produced inter-set relative SDs (RSD) of 6.1% and 5.7%, respectively.