Relation of 3-Hydroxykynurenine to the Ehrlich Diazo Reaction of Urine in Severe Tuberculosis

Abstract

IT is well known that the urines of cases of severe tuberculosis often show the diazo reaction and the urochromogen test of Weiss. Recently, we have been trying to separate and to identify the substance responsible for these reactions. The results obtained are as follows. 700 ml. of urine obtained from a severe case of tuberculosis, which gave a strong diazo reaction and the urochromogen test of Weiss, was, after being saturated with ammonium sulphate and filtered, shaken vigorously with 140 ml. of 90 per cent phenol. After standing, the separated phenol layer (115 ml.) was shaken with a mixture of 115 ml. of ether with 55 ml. of water. After standing again, we took the separated water layer, which showed a brown colour, and washed it repeatedly with ether until the ether washing showed no more colour. The water extract was concentrated in vacuo to 15 ml., in which the greater part of the substance in question was collected. Further purification was performed by chromatographing on filter paper No. 50 of the Tōyō Filter Paper Company, Tokyo, using a system of n-butanol, acetic acid and water (4: 1: 5) as the developer, ascending method. The concentrate was developed as spots on filter paper strips or wider papers (40 cm. × 40 cm.) for 20 hr. Under ultra-violet illumination, six strongly fluorescent spots appeared (A, B, C, D, E, F in diagram).