Quantitative Analysis Of Some Components In Cut Tobacco And Smoke

The investigation of the harmful components in cut tobacco and smoke is one of main task in tobacco industry.Tish is helpful for increasing aroma and reducing harm of cigarettes. In this work, the determination methods for three major polyphenols, benzo[a]pyrene, tobacco-specific nitrosamines (TSNAs) and seven volatile phenols were developed based on ultra-high performance liquid chromategraphy (UPLC) and ultra-high performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS). The results of this study are as follows:(1) A fast method based on ultra-performance liquid chromatography-photodiode array detector (UPLC-PDA) was developed for determination of scopoletin, chlorogenicacid and rutin in cut tobacco simultaneously.①The cut tobacco were extracted with 1% acetic acid water solution under ultrasonic, the extract was detected by UPLC-PDA after flitting with 0.2μm filter membrane. The polyphenols were quantified by PDA. The results showed that the RSD%(Relative standard derive) of determination of polyphenols in cut tobacco were 2.6%-4.6% with recoveries were 85.8%-91.7%.②The extraction of copoletin, chlorogenicacid and rutin in cut tobacco with microwave-assisted extraction was studied.The optimal extraction conditions of polyphenols were determined by single-factor and the orthogonal experience. The optimal conditions were as follows:methol concentration was 70%, extraction solvent temperature was 94℃,microwave extraction time was 1 min and solid to liquid ratio was 120(g/mL). The recoveries of polyphenols in cut tobacco were 96.5%-109.6%, the RSD% were lower then 5%. The effect of microwave-assisted extraction was obviously more effective compare with ultrasonic extraction. The method has advantage of simply pretreat procedure, lower time-consuming and high reproducibility, it was suitable for the fast analysis of phenolic compounds in cut tobacco.(2) A method based on ultra performance liquid chromatography with fluorescence detection (UPLC-FLD) was developed for determination of (o, p, m)dihydroxybenzene, (o, p, m)cresol and volatile phenol simultaneously. The Cambridge filter pads (CFPs) were used to collect smokes, then extracted with 1% acetic acid water solution under ultrasonic, the extract was deteced by UPLC-FLD after filtrated with 0.2μm filter membrane. The volatile phenols were quantitated and qualitated by FLD separately, and (m,p)cresol were separated completely. The results showed that the RSD% of determination of volatile phenols in mainstream smoke were 0.7%-4.6% with recoveries were 86.6%-96.9%. The method has advantage of simply pretreated procedure and high precision, it is suitable for the fast analysis of volatile phenols in mainstream smoke.(3) An accurate and simple method for the determation of Benzo[α] pyrene in cut tobacco based on UPLC-FLD was developed. The samples were extracted with cyclohexane under ultrasonic for 40 min, the extract was concentrated by evaporation plant after filter, then dissolved by menthol and acetone (V/V=10/1) and analyzed by UPLC-FLD. The mobile phase consisted of acetonitrile (85%) and water (15%), The Benzo[a]pyrene was detected at excitation wavelength 365 nm and emission wavelength 410 nm with the flow rate at 0.2 mL/min. The method showed good linearity over the range of 0.43-43.20 ng/mL, the correlation coefficient of linear equation was 0.9998. The recoveries were 94.9%-101.0%with the 3.2% of RSD%. The limit of detection (LOD) and the limit of quantity (LOQ) were 0.11 ng/g and 0.37 ng/g, respectively. This method was suitable for the qualitative and quantitative analysis of Benzo [α] pyrene in cut tobacco, due to its simpleness, sensitivity and high precision.(4) An accurate and rapid method for the determination of TSNAs in cut tobacco and mainstream smoke was developed based on UPLC-ESI-MS/MS. The pre-treatment procedure and liquid chromatogram conditions for the determination of TSNAs were compared and optimized systematically, the results showed that:Due to the different properties of NNK, NNN, NAB, NAT, no good recoveries rates of TSNAs could be achieved by solid phase extraction (SPE), and all of four TSNAs could not be measured accurately with two deuterated internal standards. The results were more accurate by detecting TSNAs directly after extraction with four deuterated internal standard.②Using 100 mM ammonium acetate,0.1% acetic acid in water and water as the extraction solvent were compared respectively, with less 5% of the RSD%, indicating that pH value had little effect on the extraction.③The column temperature affect both the peak shape of NNN, NAT. High column temperature can eliminate the bifurcation peaks, but tthis does not affect the quantification of TSNAs.④LODs of NNK, NNN, NAB, NAT were 0.10,0.26,0.22,0.08 ng/mL respectively, recoveries of them were 92.4%-99.4% in cut tobacco while were 101.7%-107.5% in mainstream smoke, RSD% were no more than 10%. This method could be used to qualitative and quantitative analysis of TSNAs in both cut tobacco and mainstream smoke.