Separation of Plant Growth Regulating Substances on Silica Gel Columns

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Abstract
By placing a tissue sample on top of a silica gel column, it is possible to extract and separate several neutral, acidic, and basic indole derivatives and unidentified naturally occurring growth regulating substances in 1 operation by a suitable choice of solvents. A bioassay is carried out in the test tubes used to collect the various fractions. The very fine particles of 100 mesh silica gel are discarded by decanting. The remaining silica gel is dried at 100[degree]C. An 8.0 sample of dried silica gel is hydrated with 5.0 ml of 0.5 [image] formic acid, slurried in the 1st eluting solvent, and packed into a glass column 1.4 cm inside diameter. Eluting solvents consist of petroleum ether (b.p. 30-60[degree]C): n-butyl alcohol saturated with 0.5 [image] formic acid. The following solvent schedule is recommended for routine survey work, using 10 ml fractions: 0.2% butyl alcohol solvent-150 ml; 3%-150 ml; 10%-100 ml; 25%-100 ml; 50%-100 ml; 75%-100 ml; 100%-100 ml. Indole, skatole, ethyl indoleacetate, and indoleacetonitrile are eluted in the 0.2% solvent; indoleacetic acid in the 3% solvent; indoleacetamide in the 10% solvent; tryptophan and tryptamine in the 75% solvent.