TOTAL HEXANE AND ETHANOL EXTRACTIVES OF TOBACCO 
by 
R. L. Stedman, W. R. Bilinsky, and A. P. Swain 
A composition study was made of the substances removed from flue- 
cured tobacco by large-scale serial extraction with n-hexane and ethanol. 
The composition of the n-hexane extractives has been described (10, 1),.1/ 
The nature and amounts of the substances in the ethanol extract anda 
material balance of'all extractives are given in this report. 
As previously described (2), 39.6 kg. of ground (10 mesh) leaf webs of 
unaged, cured Type 12 tobacco (mixed grades) were serially extracted by 
n-hexane and ethanol. After the ethanol extract was concentrated and 
refrigerated at 0° C., it gave a precipitate (EP1) and filtrate (El). An 
outline of the subsequent fractionation is given in figure 1. After removal 
of the water solubles in EPl, the water insolubles were fractionated into 
neutral and acidic and basic substances. The neutrals were separated, 
as previously outlined (9), and gave campesterol, the methyl and ethyl 
esters of higher fatty acids, and neutral resins. The acids and bases were 
not separated further. 
On continued storage at 0° C., fraction El yielded a precipitate (E2), 
which was removed by centrifugation. The precipitate showed an infrared 
spectrum identical with that of sodium propionate. It was tentatively 
identified as a salt (or mixture of salts) of propionic or other lower fae 
acids or both. To the decantate (E3) was added sufficient Skellysolve B2/ 
to give two layers (1 part E3:2 parts Skellysolve). The layers were then 
separated and to the lower layer were added sufficient ethanol (8. 47 liters) 
and Skellysolve (29.5 liters) to give two layers, and the layers were again 
separated. A further addition of the same amounts of ethanol and 
Skellysolve B to the resulting lower layer again gave two layers, which 
were separated. This final lower layer was E4,. The pooling of all upper 
layers gave E5. 
E5 was concentrated and the precipitate (E6) that formed was filtered 
off. The filtrate was evaporated to a residue, which was impregnated on 
Oy gions in parentheses refer to literature cited at the end of this report’ 
_2/Essentially n-hexane. Mention of a specific commercial product does 
not constitute endorsement by the United States Department of 
Agriculture over similar products not named. 
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