PHYSICAL AND CHEMICAL PROPERTIES OF SNAKE VENOM 89 
to lecithin chloroform, were not precipitated out by adding a large quantity 
of ether. 
Michaelis and Rona! 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, 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-hemolytic 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® a very large quantity of oleic acid 
is separated out during the preparation of cobra lecithid. 11 c.c. of 20 per 
cent lecithin solution in chloroform plus 22 c.c. of 1 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 to 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/1o NaOH, whereas 0.5079 gm. of pure oleic acid 
took 17.6 c.c. N/to 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 1 per cent venom solution. The hemolytic activity of 
the lecithid was: 0.00002 gm. dissolved 1 c.c. of 5 per cent suspension of ox 
corpuscles, but 0.coce1 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/1o NaOH to1 gm. of lecithin in alcohol, 
phenolphthalein indicator). With this lecithin hemolysin was also formed. 
But the hemolysin did not precipitate until 40 parts N/ 10 Na,COs were added 
1 Michaelis and Rona. Ueber die Léslichkeitsverhiltnisse von Albumosen und Fermenten mit Hin- 
sicht auf ihre Beziehungen zu Lecithin und Mastix. Biochem. Zeitschr., 1907, IV, 11. 
2Kyes. Bemerkung iiber die Lecithidbildung. Biochem. Zeitschr., 1908, VIII, 42. 
3 y. Dungern and Coca. Ueber Hamolyse durch Schlangengifte. Minch. med. Woch., 1907, LIV, 2317. 
