Quantitative determination of 23 flavour compounds related to potential additives in tobacco products and regulatory reporting requirements
In 2018, the National Agency of Health Oversight in Brazil (ANVISA) set forth provisions for the reporting of 165 compounds in tobacco products, to address potential attractiveness or enhanced addictiveness of these products. The objective of this study was to develop an approach for the determination of multiple compounds, suitable for ‘routine’ analysis with ‘adequate’ method performance characteristics (i.e., ‘fit for purpose’). This study describes the quantitative determination of 23 of these compounds using gas chromatography - mass spectrometry (GC-MS).
The methodology used two grams of ground tobacco product extracted with methyl tert-butyl ether (MTBE) and a saturated potassium chloride (KCl) aqueous solution. An internal standard solution containing seven deuterated compounds, was added prior to the extraction. After extraction and phase separation, an aliquot of the organic phase (MTBE) is used for GC-MS analysis. The analysis was performed using a 30 m DB-17MS column and single ion monitoring (SIM) data acquisition mode. Compound identification was done by comparing the retention times, quantifier and qualifier ratios, to those of standard reference materials. For positive identification, at least one of the qualifier ions must be within 50 - 150 % of the target ratio, and no qualifier ions below 50 % of the target ration.
Calibrations were prepared using calibration standards ranging from 0.05 µg/mL to 10 µg/mL, with a nominal internal standard concentration of 1 µg/mL. Recoveries of all target compounds from a laboratory fortified matrix (LFM) were within 79 - 119 %, with one exception for (E)-Hexen-2-al (69 %). The LFM samples were also used to evaluate method precision with the coefficients of variation (CV) for all analytes ≤ 15.
A suitable method for the quantification of 23 compounds with adequate sensitivity has been developed. However, the method does not differentiate between compounds naturally present in tobacco, those chemically bound to the matrix, and those that might have been added.