engleski

The separation of indole-3-acetic acid and related compounds in plant extracts by Sephadex chromatography

Eleven indole derivatives of biological significance were added to an extract of etiolated pea seedlings and the mixture was separated on a column of Sephadex G-15, by the following sequence of eluents: ethylenediamine-acetate, pH 4.2, ethylenediamine-acetate, pH 7.4, distilled water, and a methanol-water gradient. The bulk of plant constituents was eluted before the indolic compounds. Tryptophan and indole-3-acetic and indole-3-carboxylic acids, as well as indole-3-carboxaldehyde emerged as separate peaks. Three further peaks contained mixtures of indolic compounds easily separable by thin-layer chromatography, i. e.: 1. tryptamine and tryptophol glucoside (2-(indole-3-ylethyl)-beta-D-glucopyranoside) ; 2. indole-3-acetamide, indole-3-acetic acid methyl ester, and tryptophol ; and 3. indole-3-acetonitrile and indole-3-carboxylic acid methyl ester. Detection limits in combined Sephadex and thin-layer chromatographies ranged from 0.1 to 10 micrograms. Estimated recoveries were above 50% for indole-3-carboxaldehyde and above 80% for five other representative compounds. Thus, the described method permits the isolation, from plant extracts, of acidic, neutral, and basic indolic compounds of widely different polarity.

Sephadex chromatography ; isolation of plant metabolites ; IAA ; indole-3-acetic acid related compounds

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