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Quantitative thin-layer chromatography ofNicotianatabacumleaf surface components

Authors
Journal
Journal of Chromatography A
0021-9673
Publisher
Elsevier
Publication Date
Volume
463
Identifiers
DOI: 10.1016/s0021-9673(01)84495-2

Abstract

Abstract A thin-layer chromatographic (TLC) method and scanning densitometry technique have been developed for quantitation of major primary and secondary metabolites of Nicotiana tabacum L. leaf surface components. These components were resolved after a single development with isopropanol—chloroform—methylene chloride—hexane (7:8:6:79) using laned silica gel G TLC plates. R F values were reproducible to ± 0.01 units. Quantitation of all components of interest was accomplished by charring with 30% fuming sulfuric acid followed by densitometry using white light. Overall, charring results were semiquantitative (⩽ 10% relative error), but were quantitative (⩽ 6% relative error) for all major secondary metabolites except the sucrose esters. Quantitation of three secondary metabolites was also accomplished by scanning uncharred plates at a wavelength of 200 nm. In general, UV scanning provided semiquantitative results. For both charring and UV quantitation methods, highly correlated, curvilinear responses between mass and integration area were obtained. Advantages and limitations of these procedures versus an existing gas chromatographic procedure and their potential implementation are discussed.

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