ON THE OCCURRENCE OF CREATININ IN LEGUMINOUS SEEDS. 
^3 
Horse bean. The characteristic crystals appeared after about 3 weeks 
in the form of radiated needles or crosses. 
15. From purified syrup. 
The syrup II seemed to contain still some impurities which prevented 
more or less* the crystallisation of Creatinin zinc chloride. The syrups 
obtained from both kinds of soy bean which showed apparently strong 
color reactions of Creatinin did not show any sign of forming characteristic 
crystals of Creatinin zinc chloride under the similar conditions in whicli 
the syrups from other legumes formed the crystals. The syrups from 
soy beans evidently contained some impurities which had preventive in- 
fluence on the formation of the crystal. Starting from this point of 
view we have attemjjted first to separate Creatinin from admixed substances 
and then to try the isolation of Creatinin as its zinc chloride salt. For the 
separation of Creatinin from the syrups of leguminous seeds, we have 
adopted the following procedure. 
The alcoholic extract of the seed was concentrated to a small volume 
under reduced pressure and treated with neutral lead acetate to remove 
impurities. The excess of lead in the filtrate was removed by hydrogen 
sulphide. The lead-free filtrate was concentrated to about 30 cc. and a 
small quantity of glucose was added. The hot solution thus prepared 
was then poured into 50 cc. of the boiling Fehling's solution. The 
solution was kept boiling for 2 minutes and then allowed to cool, when a 
greyish wählte gelatinous precipitate of Creatinin cuprous oxide, mixed 
with brownish red precipitate of cuprous oxide was separated. The 
l^recipitate formed was filtered, well washed with 95 alcohol, suspended 
in water and decomposed with hydrogen sulphide. The copper sulphide 
was separated by filtration and the filtrate was concentrated to a small 
volume under reduced pressure and, with a small portion of it, color 
reactions were tested, after the complete removal of the hydrogen sulphide 
by careful evaporation, since its presence makes the Jaffe's and Wey I's 
reactions obscure. The results are shown in the following table. 
