90 



Mr. E. Ponder. The Hemolytic 



The second column gives the concentration of glycocholate in the tube if, of 

 any of these dilutions, 2 c.c. be taken, and have 2 c.c. of saline, or saline 

 solution of albumin, and also 1 c.c. of standard blood suspension, added. The 

 third column gives the value of S for such a mixture. 



In all the experiments recorded below, for convenience, quantities one-fifth 

 of these are used, the final concentrations being the same, e.g., instead of 

 2 c.c. of glycocholate plus 2 c.c. of saline plus 1 c.c. of suspension, 0'4 c.e. of 

 glycocholate, 04 c.c. of saline, and 2 c.c of suspension are used. 



(2) A solution of serum albumin in saline. The serum albumin was 

 prepared from blood, dried, and kept for some months before use. The 

 strength of the solution is 1 per cent. 



(3) A standard blood suspension, as described in the previous paper (I). 

 This is essentially a 5 per cent, suspension of thrice washed human 

 erythrocytes in normal saline. 



If to - 4 c.c. of 2 - 5 per cent, glycocholate be added - 2 c.c. of suspension, 

 and, after an interval of 5 seconds, 0'4 c.c. of serum albumin solution be 

 added, haemolysis occurs very rapidly, in about 30 seconds. It has been 

 noted above that a 1 per cent, solution of glycocholate takes over an hour to 

 produce haemolysis of this quantity of suspension. Since, after adding the 

 albumin, the concentration of glycocholate is 1 per cent., and since haemolysis 

 occurs in about 30 seconds, it is obvious that the serum albumin solution 

 has a powerful accelerating effect on the action of the bile salt. It may be 

 observed that the serum albumin solution is of itself non-haemolytie : and 

 that the rapid haemolysis is in no way explained by the fact that the blood 

 cells remain in contact with a 2 - 5 per cent, solution of glycocholate for 

 5 seconds, since the salt in this concentration will not produce any haemolysis 

 in this short time. Control experiments, using saline instead of the serum 

 albumin solution, render the accelerating action of the latter quite clear. 



The occurrence of this rapid haemolysis depends on several factors. The 

 rapid haemolysis occurs with a mixture of serum albumin, sodium glyco- 

 cholate, and blood cells. These three substances may, however, be mixed in 

 three different ways : — 



Method 1.— Put - 4 c.c. of glycocholate solution, 2 - 5 per cent., in a tube, 

 add 0"2 c.c. of blood suspension, and then, after an interval add - 4 c.c. of 

 serum albumin solution. 



Method 2. — Put - 4 c.c. of serum albumin solution in the tube, add - 2 c.c. 

 of blood suspension, and after an interval, add 0*4 c.c. of 2*5 per cent, 

 glycocholate solution. 



Method 3. — To 04 c.c. glycocholate solution add 04 c.c. of the serum 

 albumin solution, and then - 2 c.c. of the blood suspension. 



