Determination of capsidiol in tobacco cells culture by HPLC

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Authors

LITERÁKOVÁ Petra LOCHMAN Jan ZDRÁHAL Zbyněk PROKOP Zbyněk MIKEŠ Vladimír KAŠPAROVSKÝ Tomáš

Year of publication 2010
Type Article in Periodical
Magazine / Source Journal of Chromatographic Science
MU Faculty or unit

Faculty of Science

Citation
Field Biochemistry
Keywords Capsidiol Tobacco HPLC
Description Capsidiol is a bicyclic sesquiterpene, which accumulates extracellularly in plants, and has been isolated from many types of Solanaceae. It acts as a phytoalexin produced by Nicotiana tabacum in response to pathogens. Capsidiol has antifungal activity and is formed as first in tobacco and pepper plants after infestation. The amount of capsidiol in tobacco cell suspension culture has been previously determined by solid-phase extraction and organic solvent extraction with TLC or GC analysis. An HPLC method with UV detection at 210 nm on a C8 column utilizing both extraction methods was developed to analyze capsidiol in suspension cell culture. The HPLC method was linear in the concentration range of 0.1-2.0 mg/l. The lower limit of quantitation was 0.1 mg/l. Organic solvent extraction and solid phase extraction methods were compared. Both methods are generally similar in their overall efficiency (82% and 75%,respectively) but eliminations of interfering compounds are different. The relative standard deviation across five extractions of known amounts of capsidiol from plant sample was less than 5.1%. The relative standard deviation across five elicitations of cell cultures was less than 5.9%. GC-MS and LC-MS analysis of capsidiol was performed and corresponding MS spectra are presented.
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