Comprehensive analysis of lipid compounds in tobacco leaf
In recent years, lipidomics has been a vastly expanding field of research due to the development of new technologies in liquid chromatography and from recognising the role of lipids in metabolism.
Tobacco leaf consists of thousands of chemical compounds that differ in molecular weight and polarity. A comprehensive analysis combined with a statistical approach is of interest in revealing differences between tobacco samples for which further information is not available. Focusing on the higher molecular weight fraction, lipid compounds are especially of interest because they provide information on the lipidome and may help to understand changes during the curing process. However, there are only a few reports available which are related to the comprehensive and comparative measurement of lipids in tobacco leaf.
The purpose of this study was to develop a comprehensive analysis for lipid compounds in tobacco leaf.
The sample preparation consists of two steps: 1) Lipid extraction, 2) Lipid enhancement/fractionation. Lipids were extracted from tobacco leaf by the Folch method (2:1 Chloroform/Methanol), and solid-phase extraction (SPE) was performed to separate and collect fractions of neutral lipids, fatty acids and polar lipids. These three fractions were analysed using reversed-phase liquid chromatography (RPLC) with high resolution time-of-flight mass spectrometry (TOF).
As a result, more than 500 lipids could be identified in tobacco leaf. In conclusion, the entire workflow from sample preparation down to data analysis was performed in a highly automated manner.