Combined NMR and LC-MS Analysis Reveals the Metabonomic Changes in Salvia miltiorrhiza Bunge Induced by Water Depletion

Scientific Paper

10.1021/pr900995m

2010

AV III 600 MHz NMR spectrometer, micrOTOF-Q

Hui Dai,†,§ Chaoni Xiao,†,‡,§ Hongbing Liu,†,§ and Huiru Tang

Journal of Proteome Research

Abstract

Plant metabonomic analysis is essential for understanding plant systems responses to osmotic stresses.

To understand the comprehensive metabolic responses of Salvia miltiorrhiza Bunge (SMB) to continuous

and exhaustive water depletion, we characterized the SMB metabonomic variations induced by three

different drying processes using the combined NMR and LC-DAD-MS method. NMR results showed

that SMB extracts were dominated by 29 primary metabolites such as sugars, carboxylic acids and

amino acids, which were comprehensively reported for the first time, and 8 secondary metabolites

including polyphenolic acids and diterpenoids. LC-DAD-MS methods detected 44 secondary metabolites,

among which 5 polyphenolic acids together with genipin, umbelliferone and tormentic acid were found

for the first time in this plant. We found that aqueous methanol was efficient in extracting both primary

metabolites and polyphenolic acids, whereas chloroform-methanol was effective in selectively

extracting diterpenoids. We further found that air- and sun-drying markedly affected both primary and

secondary metabolisms of SMB by enhancing tanshinone and glutamate-mediated proline biosynthesis

and altering carbohydrate and amino acid metabolisms. The shikimate-mediated biosynthesis of

polyphenolic acids was promoted by air-drying but suppressed by sun-drying. These findings fill the

gap of our understandings to the metabolic responses of S. miltiorrhiza Bunge to water depletion and

demonstrated effectiveness of the combined NMR and LC-DAD-MS methods in plant metabonomic

analysis.

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