ABSTRACT
BACKGROUND: Flavor constituents play an important role in the flavor characteristics of tobacco leaves and cigarettes. Sensitive, selective, and high-throughput multi-analyte analytical methods are needed to satisfy the demand for analyzing trace-level flavor constituents in tobacco. However, trace analysis of multi-targets in a complex tobacco matrix is significantly challenging. OBJECTIVE: This study was undertaken to develop and validate a fast, selective, sensitive, and accurate GC-tandem mass spectrometry (GC-MS/MS) method for the simultaneous analysis of 250 flavor constituents in tobacco using a modified QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) extraction procedure and backflushing technique. METHODS: The samples were extracted with a mixture of acetonitrile and phosphate buffer. GC-MS/MS served as a reliable tool to quantify the flavor constituents due to its high sensitivity, selectivity, and good repeatability. RESULTS: Our evaluation showed that 243 flavor constituents presented good linearity. Average recoveries of 216 target compounds in tobacco ranged from 70 to 120% with RSDs less than 20% at three fortification levels. The limits of quantitation of 225 and 25 compounds were in the range of 2-50 and 51-112 µg/kg, respectively. This method was then successfully applied to the analysis of 15 commercial cigarette samples with different style characteristics. CONCLUSION: The modified QuEChERS method worked very well for a wide range of flavor constituents that have not been reported by QuEChERS pretreatment previously, and the use of concurrent backflushing offered significant increase in system robustness and sample throughput. The method greatly improved the detection performance and the range of the flavor constituents, and proved to be more accurate, sensitive, selective, convenient, and practical than the reported method, and thus, can be applied in routine analysis. HIGHLIGHTS: A validated QuEChERS-based GC-MS/MS method for multiple flavor constituents analysis in tobacco was reported for the first time. The use of concurrent backflushing markedly improved the instrument robustness and sample throughput.