A novel method for analyzing solanesyl esters in tobacco leaves using atmospheric pressure chemical ionization/mass spectrometer

2010 ◽  
Vol 1217 (37) ◽  
pp. 5794-5801 ◽  
Author(s):  
Naoyuki Ishida
Author(s):  
Naoyuki Ishida

Summary A comprehensive analytical method using liquid chromatography atmospheric pressure chemical ionization mass spectrometry detector (LC/APCI-MSD) was developed to determine key non-volatile components with low polarity elucidating holistic difference among tobacco leaves. Nonaqueous reversed-phase chromatography (NARPC) using organic solvent ensured simultaneous separation of various components with low polarity in tobacco resin. Application of full-scan mode to APCI-MSD hyphenated with NARPC enabled simultaneous detection of numerous intense product ions given by APCI interface. Parameters for data processing to filter, feature and align peaks were adjusted in order to strike a balance between comprehensiveness and reproducibility in analysis. 63 types of components such as solanesols, chlorophylls, phytosterols, triacylglycerols, solanachromene and others were determined on total ion chromatograms according to authentic components, wavelength spectrum and mass spectrum. The whole area of identified entities among the ones detected on total ion chromatogram reached to over 60% and major entities among those identified showed favorable linearity of determination coefficient of over 0.99. The developed method and data processing procedure were therefore considered feasible for subsequent multivariate analysis. Data matrix consisting of a number of entities was then subjected to principal component analysis (PCA) and hierarchical clustering analysis. Cultivars of tobacco leaves were distributed far from each cultivar on PCA score plot and each cluster seemed to be characterized by identified non-volatile components with low polarity. While fluecured Virginia (FCV) was loaded by solanachromene, phytosterol esters and triacylglycerols, free phytosterols and chlorophylls loaded Burley (BLY) and Oriental (ORI) respectively. Consequently the whole methodology consisting of comprehensive method and data processing procedure proved useful to determine key-components among cultivars of tobacco leaves, and was expected to additionally expand coverage that metabolomics study has ensured. [Beitr. Tabakforsch. Int. 27 (2016) 60-73]


Author(s):  
N Ishida

AbstractTriacylglycerols 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.


Author(s):  
Naoyuki Ishida

SUMMARYThere has been focus on the components with low polarity in tobacco leaf resin due to their probable relation with taste and aroma of tobacco products, the lack of a feasible analytical method and instrument has long been an obstacle to identifying the components with low polarity. The author thereby paid attention to the analysis employing nonaqueous reversed-phase chromatography hyphenated with a photo diode array detector and an atmospheric pressure chemical ionization mass spectrometry detector which has been considered applicable to the separation of significant but unknown non-volatile. The application succeeded in simultaneously separating, detecting and quantifying more than 100 non-volatile components with different low polarity such as solanesols, triacylglycerols, phytosterols, and chlorophylls. However, their compositional differences among various tobacco leaves still remained partial knowledge based on targeted analysis instead of global knowledge based on comprehensive analysis. No investigation searching for key components elucidating different tastes, aromas, species, cultivars, curing processes, and growing districts among tobacco leaves has been carried out so far. For this reason, all the quantification data were consolidated to form complete multidimensional matrix and were statistically processed to observe the categories and the key components of various tobacco leaves by principal component analysis and hierarchical clustering analysis. Tobacco leaves were first classified into three categories consisting of flue-cured Virginia, air-cured leaf, and Oriental. Solanesyl esters, phytosteryl esters and solanachromene contributed to the category of flue-cured Virginia, while air-cured leaf was characterized by free phytosterols. Oriental was featured by chlorophyll in addition to the contributory components to flue-cured Virginia. Non-volatile components with low polarity seemed to be degraded during curing process and to therefore characterize the different curing processes among various tobacco leaves.


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