A new high performance liquid chromatography - fluorescence detection method for the determination of phenolic compounds in cigarette smoke and smokeless tobacco products
A widely used method for the analysis of phenolic compounds in cigarette smoke is HPLC with fluorescence detection using acetonitrile as the mobile phase. The current global shortage of this compound prompted a search for a substitute to reduce cost and potentially improve the method. In this study, a simple, fast and sensitive HPLC method was developed using methanol instead of acetonitrile. A short (150mm, 3µm particle site) column with a pentafluorophenyl propyl (PFP) ligand in the stationary phase was chosen in order to increase separation efficiency and reduce solvent consumption. The limit of detection (LOD) and limit of quantification for the method are at the ng/mL level for most of the phenols with good linearity (R2 ≥ 0.999) and precision (RSD<10%). Mainstream yields of phenolics from 3R4F are comparable to those obtained by Health Canada method with the advantage that o, m, and p-cresols can be determined and reported separately. The results for o, m, and p-cresols under ISO conditions are 2.05 ± 0.28, 1.76 ± 0.21, and 4.23 ± 0.30 µg/cig respectively and 35.5 ± 4.72, 25.6 ± 0.87, and 51.6 ± 1.90 µg/cig in sidestream smoke. The results for phenol, o, m, and p-cresols in 2S3 smokeless reference product are 4.73 ± 0.22, 1.25 ± 0.08, 3.18 ± 0.10, and 1.68 ± 0.13 µg/g respectively.