Determination Of Flavor Additives In Tobacco Products By Chromatography-tandem Mass Spectrometry

During the past few years, flavor additives added in cigarettes have been received increasing public concern with the rapid development of cigarette production technology. Tobacco flavoring substances play an important role to solve the problem of insufficient aroma due to lower tar in the cigarette. But the aroma substances can produce complex reaction in the combustion process of cigarette. They also can produce a lot of pyrolysis products. However, some of these products have physiological toxicity. Therefore, the flavor additivesadded in cigarettes have caused increasing attention. The U. S. Food and Drug Administration(FDA), and German tobacco law have made limit regulations, so developing a method for the determination of various aroma components is very important. LC-MS/MS is a powerful tool for separating and analyzing complex samples. It has higher resolution and sensitivity than any other analysis methods. In recent years, triple quadruples mass spectrometry(Qq Q-MS/MS) has been reported to provide a much higher degree of assurance than the single stage mass spectrometry technique when determining analyses in complex matrix. The excellent selectivity and sensitivity achieved in MRM mode allowed satisfactory confirmation and quantitation for those target compounds. Additionally, the reliability and adaptability of the method were further verified by determination of linear range, recovery and reproducibility.1. An analytical method for the determination of catechol in tobacco by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS) has been developed. Sep-Park-C18 solid phase extraction cartridge was used for the enrichment of the analyte for HPLC-MS/MS analysis. The mobile phase was methanol-0.2% formic acid with gradient elution. The sample was analyzed by HPLC-MS/MS in the ESI- scanning mode with multi-reaction monitoring(MRM) for qualitation and quantitation. The linear range of calibration curve was 1.3~200 μg/kg with good correlation coefficients(r2= 0.9989). The recoveries of catechol spiked in three levels were in the range of 83.1%~98.6%, with the relative standard deviations in the range of 1.9%~5.8%. The method of HPLC- MS/MS developed in this study was initially applied to the determination of flavor additives in 6 retail tobacco samples and proved to be accurate, sensitive, convenient and practical. The analyte was found in 6 retail tobacco samples, and in some cases,the presence of quite high concentrations of catechol in tobacco should be a matter of concern.2. A new analytical method for the determination of 8 kind coumarin derivatives in tobacco by high performance liquid chromatography-mass spectrometry(HPLC-MS/MS) was developed in this paper. After ultrasonic extraction and purification, the samples were analyzed by HPLC-MS/MS in multi-reaction monitoring(MRM) mode. ESI+ scan mode is used in this method. The MS/MS fragmentation pathway of the 8 kinds of coumarin derivatives is illustrated. Methanol and 0.1% methane acid were used as the mobile phases with gradient elution. The liner ranges of calibration curves is 2~500 μg/kg with good correlation coefficients(r2 > 0.9987). The limits of detection(LODs) and limits of quantitation(LOQs) of these compounds were in the range 0.3~1.7 μg/kg and 2.0~5.2 μg/kg, respectively. The average recoveries at three spiked levels were in the ranges of 69.6%~95.1% with RSDs(n = 6) lower than 5.3%. The method is applied in the determination of 12 commercial samples.3. A method based on gas chromatography coupled to triple quadrupole tandem mass spectrometry(GC–MS/MS) was developed for the determination of 23 widely used flavor additives in tobacco products in this paper. The MS/MS fragmentation pathway of the cinnamic esters additives was illustrated. The method was defined based on two main axes, ultrasonic solvent extraction procedure with dichloromethane and analyte detection performed by GC–MS/MS in electron impact mode. The excellent selectivity and sensitivity achieved in multiple reactions monitoring(MRM) mode allowed satisfactory confirmation and quantification for the tobacco flavor additives. The linear range of the 23 flavor additives is 0.2~500.0 μg/L with good correlation coefficients(r2 > 0.9963). The limits of detection(LODs) and limits of quantification(LOQs) of these compounds were in the range 0.1~2.0 g/L and 0.4~6.0 μg/kg, respectively. The average recoveries at three spiked levels(LOQ, 2LOQ, 4LOQ) were all in the range 62.1~93.8% with RSDs(n = 6) lower than 7.8%. The method of GC–MS/MS developed in this study was initially applied to the research of flavor additives in 12 retail cigarette samples and proved to be accurate, sensitive, convenient and practical.