尿中尿素の検出のためのジアセチルモノオキシム-チオセミカルバジド・スプレー試薬

概要

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A pink spot is revealed selectively by spraying the reagent of diacetylmonoxime-thiosemicarbazide-phosphoric acid to the urea which is isolated from several urine components by either paper or thin-layer plate chromatography. After development of the chromatograms, the reagent is sprayed on the chromatograms, and then they are heated at 80°C for 30 minutes in the case of paper, or heated at 105°C for 30 minutes in the case of thin-layer plate. The identification limit of urea is 3 μg. N-Monoalkyl and N-monoaryl derivatives of urea are also detected with the reagent as pink spots at a level of 310 μg. The spray reagent is prepared by the following procedure. A thiosemicarbazide solution is prepared by dissolving 0.1 g of thiosemicarbazide in 3 ml of water by heating in a water bath and by mixing the solution with 7 ml of ethanol. A diacetylmonoxime solution is prepared by dissolving 0.3 g of diacetylmonoxime in 1 ml of ethanol and by mixing the solution with 9 ml of phosphoric acid solution which consists of 85% phosphoric acid with water in a 1 : 6 volume ratio. The thiosemicarbazide solution is mixed with the diacetylmonoxime solution, allowed to stand for 20 minutes with occasional swirling, and then filtered. The filtrate is used as the spray reagent.