Quantitative Estimation of 5‐Hydroxy‐3‐Indole Acetic Acid and 5‐Hydroxytryptophan in the Brain Following Isolation by Means of a Strong Cation Exchange Column

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Abstract
Brain tissue is extracted with perchloric acid. The pH of the extract is adjusted to 2.5 and passed through a Dowex 50 W‐X4 column. 5‐Hydroxy‐3‐indole acetic acid is eluted with 60 % aqueous methanol, and then 5‐hydroxytryp‐tophan is eluted with 0.1 M phosphate buffer, pH 6.5. Fluorometric assays are performed according to the method for 5‐hydroxytryptamine described by Andéen & Magnusson (1967). The results are satisfactory with regard to reproducibility, recovery, specificity, and sensitivity. If the above procedure is combined with that published by Atack & Magnusson (1970), then 7 different compounds can be estimated after a single column procedure.