E. V. McCollum and N. Simmonds 65 
Lot 952 A. Period 1.—Ration: Purified food mixture plus 5 per cent of 
butter fat. 
Period 2.—Ration: Same as Period 1 plus the alcohol- sane portion of 
-ether-extracted raw navy beans evaporated on dextrin and extracted with 
acetone; 7.e., alcohol-soluble, acetone-insoluble part of beans. The ration 
contained the equivalent of 30 per cent of beans. 
Period 3.—Ration: Same as Period 2 but with the alcohol-soluble, 
acetone-insoluble part of the raw beans heated 40 minutes at 15 pounds’ 
pressure in an autoclave. 
Period 4.—Ration: Same as Period 1 plus the acetone extract of the 
alcoholic extract of ether-extracted raw beans, plus the acetone-insol- 
uble residue, 7.e., the equivalent of the total alcoholic extract, after being 
heated with acetone. The ration contained the equivalent of 30 per cent 
of beans (see preparation 951 B, under Chart 3). 
In Chart 4, Lot 951 B, it is shown that growth could not be 
secured with the acetone-soluble part of the alcoholic extract of 
30 per cent of raw beans in the diet. Since 10 per cent of beans 
supplies enough of the water-soluble B to induce growth (Chart 
10, Lot 933) and neither the acetone extract or residue from the 
alcoholic extract cf 30 per cent of beans in Lot 952 A, Chart 1 
and Lot 951 B, Chart 4, induced growth, it seems certain that 
there is a slow destruction of the physiologically active substance 
in hot acetone. 
Lot 952 B. Period 1.—Ration: Purified food mixture plus 5 per cent of 
butter fat. 
Period 2.—Ration: Same as Period 1 plus 25 per cent of raw beans 
extracted with ether, then with benzene. 
Period 3.—Ration: Same as Period 2, but with the beans moistened 
with distilled water and heated at 15 pounds’ pressure for 40 minutes in 
an autoclave. 
Neither ether nor benzene removes the water-soluble B from 
beans. In the second period the rats apparently did not grow 
because of failure to eat the ration containing raw beans. After 
heating the bean residue with water and thus making it more 
palatable, growth at once took place, which shows that the 
factor B was still in the residue. 
Preparation of Extract and Residue Used with Lots 953 A and 
952 B.—1,000 gm. “of finely ground raw navy beans were ex- 
tracted sangre for 25 hours with alcohol-free ether. The 
residue was then extracted with benzene in three successive 6 
THE JOURNAL OF BIOLOGICAL CHEMISTRY, VOL. XXXIII, NO. 1 
