Abstract
A fully automated procedure for the determination of 5-hydroxytryptamine is described. The specificity and sensitivity of the assay depend upon a differential solvent extraction procedure to remove interfering 5-hydroxy-indoles and the measurement of the fluorescence of the amine after reaction with ninhydrin. The method is highly suitable for the analysis of tissue subcellular fractions since there is no interference by sucrose, and after a preliminary deproteinisation of the sample with ZnSO 4 NaOH , performed manually, the rest of the procedure is fully automated. The sensitivity of the assay depends upon the quality of the fluorimeter used but is better than 5 ng/ml of sample with a double monochromator spectrofluorimeter. There is no interference by 5-hydroxytryptophan, 5-hydroxyindole-3-acetic acid, or 5-hydroxytryptophol. The suitability of the procedure for the routine analysis of 5-hydroxytryptamine in platelet subcellular fractions prepared by zonal gradient centrifugation has been explored and the soluble phase and particulate bound amine stores can be identified.
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