Abstract
Triacylglycerols in tobacco leaves were investigated to further understand their compositional and metabolic differences among various tobacco leaves and their changes during the curing processes. Non-aqueous reverse-phase chromatography for efficient separation of triacylglycerols and atmospheric-pressure chemical-ionization mass spectrometry for their detection were selected for quantitative analysis. For measuring their absolute amounts, authentic triacylglycerols comprised of the same fatty acids were introduced to generate calibration curves and to quantify the analytes. A validation study using the combination of appropriate separation and authentic triacylglycerols produced very good results in terms of linearity (> 0.999), limit of detection (3-4 ng), limit of quantification (10-15 ng) and recovery ratio (80-120%). The quantification of triacylglycerols in tobacco leaves and a subsequent principal component analysis clarified for the first time their compositional and metabolic differences among various tobacco leaves.