Extraction, Purification, and Identification

The chromogenic reagent described has been tested with seventy-nine indole derivatives and found to be very sensitive and indole-specific. The lower limit of detection on silica gel thin-layer plates was between 25 and 50 ng for most indoles. Phenols and hydroxy-, and amino-benzoic acids, hydroxy-, and methoxy-cinnamic acids did not yield chromophores with the exception of p-amino-benzoic acid and p-hydroxy-cinnamic acid which gave yellow and pink chromophores at concentrations greater than 1 and 2 mug. Although many of the C-3 substituted indoles such as indole-3-acetic acid and tryptamine had colors in the reddish-violet-blue color region, most exhibited sufficient color differentiation to allow their identification by thin-layer chromatography. The procedure was simple and required only 10 min from the time of spraying the thin-layer plate until full color development was reached. The colors had a wide spectral range from yellow of the indole-3-glyoxylamide chromophore to blue of the melatonin chromophore, and were extremely stable.