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small quantities of lime salt on the lipolysis is not hard to test: 
Some drops of commercial olive oil are mixed with highly diluted 
soda and a few drops of lipase. After thorough intermixture by which 
the enzyme, left in solution by the weak alkali, is equally distributed 
in the fluid, and after addition of a little phenolphthalein, equal 
portions of the emulsion are put in two tubes, after which to the 
one calcium is added, for instance 1 drop of CaCl, 1°/, to 5 ec. 
of the fluid. The red colour will disappear, at least will get much 
fainter. Subsequently the fluid is made as red again as that of the 
other tube by cautious addition of sodium carbonate. When both 
tubes are heated to the temperature of the body, it will be seen that 
the one with calcium soon loses its red colour while its acidity is 
gradually increasing, whereas the colour of the other hardly changes 
or does not do so at all, in an hour’s time. The reaction should be 
made very slightly alkaline, because the enzyme, especially at the 
temperature of the body is soon destroyed by alkali. 
It thus appeared that, in order to confer activity on the almost 
inactive lipase, the only salt in it being a little sodiumcarbonate, 
we do not want the addition of the mixture of substances dissolved 
in water from the pancreatic extract, but that calcium chloride will 
do for the purpose. 
For a more exact investigation of the lipolysis 1 proceeded as 
follows: 3 to 4 ec. of the lipase was mixed with about double the 
quantity of a 0.2°/, solution of Na,CO, and on addition of ten drops 
of phenolphthalein diluted with water to 200 cc. This slightly opa- 
lescent fluid of reddish coloration was equally distributed among 
four bottles of 150 cc. capacity each; to each bottle 1 cc. of neutral 
olive oil was added, the oil being liberated from fatty acids by 
shaking up the ether solution with sodium hydrate. Beforehand the 
bottles were furnished with the substance whose action on the lipase 
was the object of our research. When OH-ions were fixed by the 
investigated matter, the pink colour was equalized in all the bottles 
by means of Na,CO,. Hereupon the well-corked bottles were put 
in the thermostat at 38° C. and turned slowly round an horizontal axis 
mostly for 6 hours, so as to ensure a constant regular intermixture 
of their contents. After 50 ce. of 92 °/, alcohol had been added to each 
n 
bottle, the amount of acid was determined by titration with z Nao. 
It now invariably appeared, that some acid had also been set free 
in the bottles containing only lipase, some sodium carbonate, water 
and oil. The quantity varied in different preparations of the enzyme, 
but was the same in each preparation on different days. It can hardly 
