9 8 



Scientific Proceedings (57). 



Immunization experiments were successfully carried out, 

 increasing doses of I to 15 c.c. being administered. Dogs survived 

 such doses, and developed agglutinins to B. pyocyaneus. 



In animals which survived ten days or longer, areas of un- 

 resolved pneumonia and thickened pleura were the rule. 



58 (875) 



A method for the separation of lipins from lipin extracts. 



By Jacob Rosenbloom, M.D., Ph.D. 



[From the Biochemical Laboratory of the Western Pennsylvania 

 Hospital, Pittsburgh, Pa.] 



Owing to the solubility of lipins in ether and alcohol we have 

 a basis for the isolation of these substances. To separate them 

 from their ether and alcohol solution we have several methods. 

 As a rule these methods are expensive as most of the solvent is lost, 

 they require considerable time and in some of the methods heat is 

 applied, a bad procedure on account of the labile nature of some 

 of these substances. 



I have devised a method which gives good results, allows the 

 solvent to be regained and takes little time without using heat. 

 It is based on the fact that lipins are insoluble in water. The 

 method is as follows; to the ether or alcohol extract of the lipins 

 add cold water containing 0.5 per cent, of sodium chloride, 1 

 till no further precipitation occurs. The water should be added 

 slowly without shaking or stirring, otherwise some of the lipins 

 will be emulsified. Any of the precipitate not coming to the 

 surface may be obtained by filtration. 2 It will be found that this 

 precipitate contains the lipins and if one wants to obtain the 

 phospholipins from the precipitate, simply wash thoroughly with 

 acetone until no residue is obtained, when the washings are evapo- 

 rated to dryness. The acetone removes all the lipins except the 

 phospholipins. 



1 The addition of sodium chloride to the water helps .to flocculate the lipins and 

 by raising the specific gravity of the solution, allows the lipins to come to the surface, 

 where they may be skimmed off by means of a spoon. Percentages of sodium 

 chloride under 0.5 per cent, do not give good results. 



2 The solvent can now be obtained by distilling it from the filtrate. 



