156 J. M. HAMILL. 



on a water-bath and the residue extracted with ether. The ethereal 

 extract was then evaporated and the residue consisting of crystals of 

 cholesterin was weighed. From 4'320 gms. of ether extract '225 gms. 

 of cholesterin were obtained, i.e. 5'2 gms. of cholesterin per 100 gms. of 

 ether extract. 



No reducing sugar could be detected in the chyle ; Fehling's 

 solution showed no reduction when heated with the chyle itself or with 

 the filtrate obtained from it after the addition of trichloracetic acid. 



Enzymes. Grohe (10) , and later Hensen' 15 ' have demonstrated the 

 presence of amylase in the chyle; Munk and Rosenstein (28) , in the 

 case which they examined have confirmed these authors observations. 

 The method I employed was as follows : The chyle was filtered under 

 pressure through a clay filter, the operation was a slow one owing to the 

 readiness with which the fat in the chyle blocked the pores of the filter. 

 Eventually, however, a slightly opalescent filtrate was obtained. Two 

 test-tubes, each containing 1 c.c. of a boiled starch solution, were taken ; 

 3 c.c. of chyle filtrate were added to one and 3 c.c. of boiled chyle filtrate 

 to the other. Both tubes were kept at 37 C. with toluol for 15 hours. 

 At the end of this time the boiled tube gave no reduction when 

 heated with Fehling's solution whereas the unboiled tube gave a 

 heavy precipitate of cuprous oxide.. 



The presence of lipase in the filtrate was proved by two different 

 methods : (1) Two test-tubes each containing 3 c.c. of the filtered 

 chyle were taken; one of these was boiled and served as a control. 1 c.c. 

 of neutral olive oil was then added to each tube and both were kept in 

 the presence of toluol for 15 hours. At the end of this time the boiled 

 tube was found to be still neutral whereas the unboiled tube required 



N 

 1 c.c. of y^NaOH to neutralise it. The second method employed 



depends upon the fact that amyl salicylate is split by a lipase into amyl 

 alcohol and salicylic acid, which latter gives a violet coloration with a 

 drop of ferric chloride. Amyl salicylate is for this purpose superior to 

 other esters since it is very stable and does not easily hydrolyse in the 

 presence of water alone. Two test-tubes each containing 1 c.c. of amyl 

 salicylate were taken ; 3 c.c. of the filtered chyle was added to one tube 

 and 3 c.c. of the boiled filtered chyle was added to the other as a control. 

 The tubes were then kept at 37 C. for 15 hours in the presence of toluol. 

 At the end of this time a drop of ferric chloride was added to each of 

 the tubes. No alteration was observed in the boiled tube but in the 

 other a deep violet coloration was produced. 



