E. V. McCollum and N. Simmonds 67 
Period 3.—Ration: Same as Period 2 but with alcohol-soluble, benzene- 
insoluble residue of raw beans heated 40 minutes in an autoclave at 15 
pounds’ pressure. 
Period 4.—Ration: Same as Period 3 but with the benzene-soluble por- 
tion of the alcohol-soluble part of the beans included; 7.e., the total alco- 
holic extract of beans after treatment with benzene (used extract equiva- 
lent to 30 per cent of beans). 
The alcoholic extract of raw navy beans contains the dietary 
essential water-soluble B. Although benzene does not extract 
this substance from the beans directly (Chart 1, Lot 953 A, 
Period 2; Chart 2, Lot 994 B), once it has been removed from 
the beans by means of hot alcohol, and the alcohol evaporated, 
<y 7 ” (Oa 
ial a Zs 
benzene dissolves the physiologically active substance fairly 
readily (Chart iyo Vbtahe 953 A, Period 3; Chart 2, Lot 953 B, 
Period 4). 
Lot 994 A. Period 1,—Ration: Purified food mixture plus 5 per cent of 
butter fat. 
Period 2.—Ration: Same as Period 1 but with the benzene extract of 
ether-extracted cooked navy beans, equivalent to 30 per cent of beans in 
the diet. 
Benzene does not take out the water-soluble B from the beans 
directly (Chart 1,.Lot 953 A, Period 2). It must be first re- 
moved by hot alcohol, after which it is fairly soluble in hot ben- 
zene (compare Chart 1, Lot 953 A, Periods 2 and 3, and Chart 2, 
Lot§.953 B, Period 4). Lot 994B, Period 2, shows that the 
