Spermine, Choline, Ergothioneine 163 



Not only was he able to refute the mistaken belief in the identity 

 of spermine and diethylenediamine but he was also the first 

 to analyse correctly the gold salt and the chloroplatinate of sper- 

 mine and to establish that 'the organic base which is at the bottom 

 of these double salts would have the composition, C5H14N2 (1898, 

 1906). 



However, not until 1924 was conclusive chemical and crystallo- 

 graphic evidence brought forward to prove the identity of spermine 

 isolated from semen with the base obtained by similar methods from 

 various animal organs and also from yeast. Credit for this is due to 

 Otto Rosenheim (1924). In 1924, Dudley, Mary Rosenheim and O. 

 Rosenheim in England, and Wrede in Germany, concluded from 

 the molecular weight estimations of benzoylspermine and m-mixo- 

 benzoylspermine, respectively, that the molecular formula of sper- 

 mine is C10H26N4, and not as formerly assumed C5H14N2. 



Rosenheim and his colleagues obtained spermine in the free state 

 as a crystalline, optically inactive substance which melts between 

 55° and 60°, and distills at about 150° in vacuo without decomposi- 

 tion. They also found that the base is stable in hot concentrated 

 alkali and in boiling hydrochloric acid. Their relatively simple 

 method of isolation depends on the steam-distillation of spermine 

 from a strongly alkaline solution. 



Derivatives of spermine and their use in forensic medicine 



There are several well-defined compounds of spermine, a list of 

 which is given in Table 22. Apart from the highly characteristic 

 insoluble phosphate, spermine can be identified particularly easily 

 as a picrate, which can be prepared from the free base, the phos- 

 phate, or directly from semen. Spermine picrate, like the phosphate, 

 is extremely insoluble in water. Crystallographic analysis has shown 

 that it is identical with the substance responsible for the so-called 

 Barberio reaction, a chemical test of diagnostic value in forensic 

 medicine. Barberio's (1905) test consists in the addition to semen, 

 or to an aqueous extract from the seminal stain, of picric acid in 

 concentrated aqueous or ethanolic solution; in the presence of picric 

 acid, there follows within a few minutes, the formation of abundant 

 yellow crystals, resembling in shape the crystals of spermine phos- 

 phate. The statement by Barberio that the reaction appears to be 

 12 



