89 



to lecithin chloroform, were not precipitated out by adding a large quantity 

 of ether. 



Michaelis and Rona 1 add further knowledge as to the mechanism of lecithid 

 formation. These authors found that chloroform solution of mastix when 

 shaken with aqueous solution of rennet takes up a certain part of this enzyme 

 as chloroform-alcohol solution forms and can equally be precipitated out by 

 means of ether, an exact parallel phenomenon to the process of lecithid prepa- 

 ration of Kyes. 



Landsteiner and Jagic and Michaelis and Rona are inclined to regard the 

 phenomenon as of merely physical nature, namely, colloidal reaction. 



Kyes, 2 however, does not consider the physical explanation of Michaelis 

 and Rona of the mastix-rennet phenomenon applicable to the formation of 

 venom lecithid, inasmuch as there is a very great and important difference 

 between these two sets of superficially analogous processes. Venom lecithid 

 does not contain any trace of the original materials from which it has been 

 derived. No evidence of free venom-haemolytic amboceptors or of native 

 lecithin can be brought out. Its chemical and physical properties are quite 

 different from the original materials. On the other hand, the precipitate of 

 mastix-rennet shows at least that the ferment is intact in all its original charac- 

 teristics. Here it has been a mere physical process, not comparable with 

 the venom-lecithid formation. 



According to von Dungern and Coca 3 a very large quantity of oleic acid 

 is separated out during the preparation of cobra lecithid. n c.c. of 20 per 

 cent lecithin solution in chloroform plus 22 c.c. of i per cent cobra venom 

 solution in 0.8 per cent NaCl solution were shaken 2 hours; 5 times volume of 

 ether; precipitation of lecithid. From 2 gm. of lecithin about 1.206 gm. 

 lecithid were obtained. 



From 10 c.c. chloroform solution 0.5642 gm. of pure acid oil went into 

 ether. Its acidity agreed with that of oleic acid. 0.5079 gm. of this oil was 

 dissolved in absolute alcohol and then determination of acidity was made. 

 It required 16.5 c.c. of N/io NaOH, whereas 0.5079 gm. of pure oleic acid 

 took 17.6 c.c. N/io NaOH. Baeyer's double bond test with permanganate 

 was positive; lead salt perfectly soluble in ether. In the separated saline solu- 

 tion there was still cobra venom, which, when used in 5 times the original, 

 dissolved the blood in the presence of 0.5 c.c. of 0.05 per cent lecithin just as 

 rapidly as the original i per cent venom solution. The haemolytic activity of 

 the lecithid was: 0.00002 gm. dissolved i c.c. of 5 per cent suspension of ox 

 corpuscles, but o.ooooi gm. did not act. 



A preparation of lecithin purchased from E. Merck they found to contain 

 a large amount of acid (5.5 c.c. N/io NaOH to i gm. of lecithin in alcohol, 

 phenolphthalein indicator). With this lecithin haemolysin was also formed. 

 But the haemolysin did not precipitate until 40 parts N/io Na 2 CO 3 were added 



1 Michaelis and Rona. Ueber die Loslichkeitsverhaltnisse von Albumosen und Fermenten mit Hin- 



sicht auf ihre Beziehungen zu Lecithin und Mastix. Biochem. Zeitschr., 1907, IV, n. 

 J Kyes. Bemerkung iiber die Lecithidbildung. Biochem. Zeitschr., 1908, VIII, 42. 

 a v. Dungern and Coca. Ueber Hamolyse durch Schlangengifte. Munch, med. Woch., 1907, LIV, 2317. 



