Metabolic Profiling of Chinese Tobacco Leaf of Different Geographical Origins by GC-MS

Abstract

Tobacco leaf obtained from different geographical areas in China was profiled using gas chromatography-mass spectrometry (GC-MS) coupled with multivariate data analyses. Hierarchical cluster analysis (HCA) and principal component analysis (PCA) showed that the tobacco metabolome was clearly dependent on geographical origins; climatic conditions, such as temperature and precipitation, imposed a greater impact on metabolite levels than the cultivars. By orthogonal partial least-squares-discrimination analysis (OPLS-DA), 20 metabolites that contributed to the discrimination were screened, including primary metabolites (sucrose, D-fructose, D-mannose, D-glucose, inositol, maleic acid, citric acid, malic acid, L-threonic acid, L-proline, L-phenylalanine), secondary metabolites (chlorogenic acid, α- and β-4,8,13-duvatriene-1,3-diol, nicotine, quinic acid), and four unknown metabolites. The results suggest that metabolic profiling using GC-MS combined with multivariate analysis can be used to discriminate tobacco leaf of different geographical origins and to provide potential indicators of tobacco origins.

Genetic variation in alkaloid accumulation in leaves of Nicotiana

“…Different from our results, Lin et al (2002) found that the total alkaloids contents of upper, middle, and lower leaves of plants grown in Cuibiyihao in Fujian Province, Southeast China, were 51.35, 41.19, and 28.06 mg/g, respectively. Yunnan Province is located in Southwest China and is influenced by a low latitude plateau, mountainous terrain, a monsoon climate with strong ultraviolet light, long total sun exposure time, relatively stable average temperature, and distinct, alternating dry and rainy seasons (Zhang et al, 2013). The huge differences between locations in temperature, illumination, water, and soil conditions might be responsible for the large differences found in the alkaloid contents from the same cultivars and leaf positions.…”

A Simultaneous Analytical Method to Profile Non-Volatile Components with Low Polarity Elucidating Differences Between Tobacco Leaves Using Atmospheric Pressure Chemical Ionization Mass Spectrometry Detection

“…The term is nowadays described as a methodology which "seeks to identify and quantify the complete set of metabolites in a cell or tissue type and to do so as quickly as possible and without bias" (7). Metabolomics was therefore expected to become a useful methodology for tobacco leaves and has already shown the usefulness to correlate differences in components with growing districts within a nation (8,9) and with differences resulting from systematic acquired response (SAR) caused by numerous natural stresses (10,11). However, the methodologies applied to those studies has employed only gas chromatography mass spectrometry detection (GC/MSD) oriented to volatile components (8,9), or liquid chromatography electrospray ionization mass spectrometry detection (LC/ESI-MSD) for components with middle to high polarity (11).…”

“…Metabolomics was therefore expected to become a useful methodology for tobacco leaves and has already shown the usefulness to correlate differences in components with growing districts within a nation (8,9) and with differences resulting from systematic acquired response (SAR) caused by numerous natural stresses (10,11). However, the methodologies applied to those studies has employed only gas chromatography mass spectrometry detection (GC/MSD) oriented to volatile components (8,9), or liquid chromatography electrospray ionization mass spectrometry detection (LC/ESI-MSD) for components with middle to high polarity (11). Since non-volatile components with low polarity such as solanesols (12,13), chlorophyll derivatives (14), triacylglycerols (15), solanachromene (16) and phytosterols (17) (Figure 1) are also included in tobacco leaf and have been considered unsuitable for detection in using the previously employed instruments (8)(9)(10)(11), the metabolomics applied to tobacco leaves has yet to cover all the components in tobacco leaves.…”

“…However, the methodologies applied to those studies has employed only gas chromatography mass spectrometry detection (GC/MSD) oriented to volatile components (8,9), or liquid chromatography electrospray ionization mass spectrometry detection (LC/ESI-MSD) for components with middle to high polarity (11). Since non-volatile components with low polarity such as solanesols (12,13), chlorophyll derivatives (14), triacylglycerols (15), solanachromene (16) and phytosterols (17) (Figure 1) are also included in tobacco leaf and have been considered unsuitable for detection in using the previously employed instruments (8)(9)(10)(11), the metabolomics applied to tobacco leaves has yet to cover all the components in tobacco leaves. In the first place, comprehensive analyses for metabolomics consist of a separation and detection part which are hyphenated so as to maximize comprehensiveness of method (7,(18)(19)(20).…”

“…In the first place, comprehensive analyses for metabolomics consist of a separation and detection part which are hyphenated so as to maximize comprehensiveness of method (7,(18)(19)(20). Gas chromatography (GC) for volatile components (8,9,21), liquid chromatography (LC) for components with low to high polarity (11,22) and capillary electrophoresis (CE) for hydrophilic components (23) have therefore been selected in comprehensive analysis according to the chemical characters of target components. Detection for comprehensive analysis in using chromatography has basically been accomplished by mass spectrometry detection (MSD).…”