Background: Manufacturers of tobacco products in Belgium are legally required to register the exact composition of their products. In the context of this bachelor's thesis, a chromatographic fingerprinting method was developed to produce unique fingerprints of tobacco varieties. This method aims to enable the verification of declared compositions and contributes to the detection of potential fraud or evasion of regulations.
Aim: The aim of this bachelor's thesis was to develop a fingerprinting method to distinguish six tobacco varieties using ultra-high-performance liquid chromatography (UHPLC) with diode-array detection (DAD). Additionally, the study examined whether this method could be transferred to a UHPLC-system with mass spectrometry as the detection technique.
Method: During method development, various parameters were tested and optimized, including the pH and organic component of the mobile phase, the gradient, column type and the influence of different extraction solvents. After analyzing the obtained chromatograms, the most suitable method was selected and further optimized. The method was then applied to mixtures of tobacco varieties prepared through trituration. Finally, the method was used with mass spectrometry for a more detailed analysis of tobacco mixtures.
Results: The method produced largely unique, stable and reproducible chromatograms, allowing the six studied tobacco varieties to be separated from one another. Flue-Cured and Oriental tobacco, in particular, showed clear differentiation. The application of mass spectrometry gave more peaks and visual distinctions between varieties, although further optimization is necessary.
Conclusion: The developed fingerprinting method is suitable for distinguishing the studied tobacco varieties, both in pure form and in mixtures. With further optimization of chemometrics and mass spectrometry, the method has potential for application to commercial tobacco products.
