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1. Pregnane-3:17:20-triols give a blue fluorescent spot after heating a chromatogram sprayed with approximately 25% trichloracetic acid in ethanol. A spot containing 2 µg may be detected by this reaction. Out of forty-eight steroids examined only atiocholanediol gave a similar fluorescence. Certain other steroids give different characteristic colours and fluorescence.

2. The test has been successfully used to detect pregnanetriol in the non-ketonic fraction of urine extracts from a normal male and from a patient with adrenal hyperplasia. It has been used to detect the production of pregnanetriol by in vitro reduction of pregnane-3:17-diol-20-one.

3. The identity of the substances which gave this reaction and which were obtained from urine and from in vitro reduction was confirmed by isolation and characterization.