Urobllinogenに関する研究 第二編 StercobilinogenとMesobilinogenのEhrlich氏Aedehyde反応呈色物質のChromatographyによる分離にて

The paper chromatography was attempted for the separation of stercobilinogen, predominating in urobilinogen of living body, and mesobilinogen, being supposed to partly exist in vivo, on the coloured substances of them by the addition of the Ehrlich's aldehyde reagent, and the columnchromatography with the column of a cellulose powder was also attempted for the separation of the urobilin coexisting in the coloured substances of urobilinogen by the addition of the Ehrlich's aldehyde reagent. And the results are as follows. 1. The coloured substance of urobilinogen and urobilin are definitely separated, but the separation between the coloured substances of stercobilinogen and mesobilinogen by the addition of the Ehrlich's aldehyde reagent is impossible, by the paper chromatography with the developing solvents of a methanol and water mixture (1:2), a methanol, butanol and ammouia mixture (1:2:1), a 28 % amonia and butanol mixture (4:96), N-Ammonia-condensed butanol, 10 % Ammoniacondensed butanol, and a butanol, and a butanol, propanol and ammonia mixture (2:2:1) etc. 2. The coloured substances of stercobilinogen and mesobilinogen with the Ehrlich'c aldehyde reagent display the Rf 0,81 and 0. 64 respectively and are definitely separated, on the use of the paper with the stationary phase made with glacial acetic acid and, with the developing solvents of a methanol, ammonia and water mixture (76:4:20) and a methanol, butanol and ammonia mixture (2:2:1). 3. The separation of stercobilin and urobilin IX α are easily obtained by the paper chromatography with the developing solvents of a methanol, ammonia and water mixture (76:4:20), and a methanol, butanol and ammonia mixture (2:2:1). 4. The coloured substances of urobilinogen with the Ehrlich's aldehyde reagent and urobilin in them can be separated by the column chromatography with the column of cellulose powder and the developing solvent of a methanol and water mixture (1:2). Therefore, as the absorption curve of the coloured substance with the Ehrlich's aldehyde reagent displays having only one absorption maximum at 558 mμ, it must he definitely distinguished from the urobilin which becomes coexisted during the process.