Germination of Heliant hits annnus. 705 
The material obtained in this manner was not oxidized in the least and 
was a white or yellow powder. 
Upon the material thus prepared duplicate analyses were made for the 
following constitutents: ash, moisture, ether extract, total sugar, reducing 
sugar, cellulose, total nitrogen, and proteid-free nitrogen. The amount of 
free fatty acid and the iodine number are determined for the ether extract. 
The determinations of most of these constituents were made by the 
ordinary official methods of the American Association of Official Agri¬ 
cultural Chemists. In order to obtain the amount of protein, the protein 
nitrogen was multiplied by the factor 5-5, a factor which is based upon the 
work of Osborne and Campbell 1 upon the proteid of the sunflower seed, 
which showed it to contain 18 % of nitrogen. 
For the estimation of the proteid-free nitrogen the following method 
was used:— 
A weighed portion of the finely ground material was extracted with 
ether and then stirred up with a definite volume of cold water and allowed 
to stand with frequent stirring for one half-hour. At the end of this period 
the mixture was heated to boiling, a few drops of acetic acid added, and the 
boiling continued for several minutes. The mixture was then filtered, and 
to an aliquot portion of the filtrate an equal volume of a solution of 7 grams 
tannic acid and 2 c.c. of glacial acetic acid per 100 c.c. was added. The 
tannin precipitate of the protein was then filtered and an aliquot portion 
of the filtrate taken for the nitrogen determination. 
The iodine number was determined by the Hanus method. 
The result of these analyses are expressed in Table I. The figures in 
any one column of that table represent the averages of the duplicate 
analyses made. In order to be certain that the results obtained in the 
analyses of the seedlings at any given stage were indicative of the processes 
normally taking place at that period, a second lot of seedlings was grown, 
reproducing the conditions as nearly as possible, and duplicate analyses of 
these made. Thus in Table I the figures in column a , Stage I, represent 
the average of the duplicate analyses on the hypocotyls or cotyledons for the 
first lot of seedlings. The figures under b for the same stage represent the 
average of the duplicate analyses on the hypocotyls or cotyledons of the 
second lot of seedlings grown. In Stages I and II a third lot was grown 
and analysed for certain constituents as shown. Two series of duplicate 
analyses were made on the cotyledons of the seeds as shown in the table, 
thus ensuring against possible error in sampling. 
It is seen that the results obtained from the two lots of seedlings of 
any one stage correspond as closely as could be expected. It was found 
that under approximately like conditions, the time required for the different 
lots of seedlings to reach the desired stage varied only a few hours. 
1 l.c. 
