Multi-analyte approach for determining the extraction of tobacco constituents from pouched snus by consumers during use

Abstract

BackgroundSnus is a smokeless oral tobacco product with a significant history of use in Sweden, where it is regulated under food legislation. Users place a small porous sachet or a pinch of loose snus between the upper jaw and cheek for approximately one hour, leading to partial intake of tobacco constituents. To understand user exposure to tobacco, a multi-analyte approach based on the extraction of pouches by methanol, ethanol and water was validated and applied to the measurement of various constituents, including nicotine, four tobacco-specific nitrosamines (TSNAs), propylene glycol, water, ammonium, nitrate, sodium, chloride, linalool, citronellol, linalyl acetate and geraniol, extracted from snus pouches during use by human consumers.ResultsAfter validation against established single-analyte methods, the multi-analyte approach was used to determine constituent levels in snus pouches before and after one hour of use. Although the concentrations in the snus pouches varied from nanogram (e.g. TSNAs) to milligram (e.g. nicotine, sodium and propylene glycol) quantities (25.1 ng to 35.3 mg per 1 g pouch), the mean percentage extracted varied only from 19.2% for linalyl acetate to 37.8% for the TSNA 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) among all constituents analyzed. The TSNAs, some of which are known carcinogens, showed the highest percentage extraction (range 34.6%–37.8%). Measurement variability was low for all analytes, ranging from 2.4% (total TSNAs, NAT) to 9.5% (geraniol). By contrast, inter-subject variability ranged from 6.7% (NAB) to 52.2% (linalyl acetate), and was greater than 20% for eight of the constituents analyzed. Intra-subject variability ranged from 3.4% (citronellol) to 29.7% (geraniol).ConclusionsGenerally, less than a third of each constituent tested was extracted during one hour of snus use, independent of constituent concentration. The variable nature of in-use extraction was shown to be driven by inter-subject variability. The results provide insight into possible mechanisms controlling constituent extraction in the mouth during snus use, and provide reference data for the development of in-vitro laboratory systems for estimating extraction of tobacco constituents from snus.

Analysis of Tobacco-Specific Nitrosamines in the Common Smokeless Tobacco Afzal in Oman

“…21 A recent study revealed that the total TSNA level in an unused 1.000 g pouch of snus was approximately 0.830 µg/g, with quantifiable levels of NAT, NNK, NNN and NAB (0.268 µg/g, 0.191 µg/g, 0.344 µg/g and 0.025 µg/g, respectively). 33 Therefore, total TSNA levels in the Afzal sample analysed in the current study were fivefold higher than the recommended GOTHIATEK ® limits. Specific TSNA levels in Afzal were six-fold, 2.4-fold, 5.2-fold and 8.4-fold higher than the NAT, NNK, NNN and NAB levels, respectively, in snus.…”

“…TSNAs are highly water-soluble; therefore, as the moisture content of a STP increases, the levels of soluble TSNAs may also rise, resulting in enhanced absorption by the oral mucosa. 33 The authors of the current study strongly believe that TSNAs are not safe at any level. As a result, it is strongly recommended that existing legislation in Oman regarding the illegal sale of Afzal be more strictly enforced.…”

“…Moreover, Afzal users may be exposed to higher levels of TSNAs as a result of varying self-determined pinch sizes; pinch sizes of Afzal are usually greater than the portions provided in packets of the more monitored and controlled Swedish snus. 12,13,33,34 It is important to bear in mind that the results obtained by the current study and previous research on Afzal were derived from a single package of Afzal blend. 12,13 The findings of future studies may therefore differ due to variations in the additives and manufacturing processes used.…”

Characterisation of Nicotine and Cancer-Enhancing Anions in the Common Smokeless Tobacco Afzal in Oman

“…7,19 Higher nicotine levels are found in moist snuff in comparison to the dry form. 1 Additionally, it has been found that American moist snuff contains higher levels of NO 2 -and Cl -than the dry type.…”

“…2 The reported Cl -content in unused snus (35,300.00 µg/g) was close to that of Afzal as observed in the current study (33,170.80 µg/g). 19 Quantities of Swedish snus are well monitored as they are typically sold in 1.00 g tea bag-sized sachets; 19 in contrast, Afzal users usually self-determine doses (typically a pinch-sized amount per single dose ranging from 1.00-2.00 g) which are then repeated as desired. 13 As a result, Cl -consumption will increase due to repeated Afzal use during the day.…”