A NEW STANDARD FOR USE IN THE COLORIMETRIC

DETERMINATION OF IODINE.
BY ATHERTON SEIDELL.

(From the Division of Pharmacology, Hygienic Laboratory, Washington, D. C.)

(Received for publication, July ~9, 1907.)

Of the methods which have been proposed and used for the
determination of small amounts of iodine in such substances as
thyroid glands, blood, muscle, and various animal and plant

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substances the one which has apparently been found the most
satisfactory is that adapted by Baumann and Roosl from the
old process of Rabourdin and at present known as Baumann’s
method. According to this process the weighed sample of mate-
rial which should contain not more than a few milligrams of
iodine is moistened with water and mixed thoroughly with about
five times its weight of powdered sodium hydroxide. After
warming gently the heating is increased until the mass fuses to
a liquid or semi-solid paste. If much charred organic matter
is present a little sodium nitrate may be added for its more com-
plete oxidation, and the heating continued until a clear melt is
obtained. After cooling, water is added and the aqueous solu-
tion containing all of the iodine in the form of an inorganic salt
is filtered into a separatory funnel. The solution is acidified
with sulphuric acid while being kept cool and is then treated with
IO--20 drops of I per cent aqueous sodium nitrite solution and
the liberated iodine shaken out with chloroform. The intensity
of the pink color in the chloroform layer is proportional to the
amount of iodine present, and it is only necessary to compare this
color with standards of known iodine content in order to ascertain
the amount of iodine in the unknown sample. The standards so
far used have been prepared from aliquot portions of standard
potassium iodide solution, shaking out the liberated iodine with
the same volume of chloroformused for the unknown sample.

lZeit.dw. f. pkysiol. Chem., xxi, p. 489, ‘895-96.

30 1
392 Calorimetric Iodine Determination

The experience of Baumann and of many other investigators,

has shown that the pink colors in the chloroform layers fade
rapidly and therefore it is absolutely necessary that the stand-
ards be freshly prepared for practically all determinations.
Many efforts have been made in this laboratory to prevent or
even retard the fading of the iodine color in the chloroform but
without success. In endeavoring to overcome this difficulty
in another way, an examination of the tints of the aqueous solu-
tions of a large number of pink dyes was made, in the hope of
finding one, which resembled near enough the color of the chloro-
form solution of iodine, to be used as a standard for the colori-

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metric estimation of the latter. Although none of the several
dozen pink dyes which were examined gave solutions possessing
a tint matching exactly that of iodine dissolved in chloroform,
several gave colors which corresponded near enough for all prac-
tical purposes. Among these may be mentioned Neutral Red,
Rubin (patent acid) and Fuchsin S (acid fuchsin according to
Weigert), all of which dyes in the present case bore the labels
of Dr. G. Grubler & Co., Leipzig. The Fuchsin S was, on the
whole, found to be the most satisfactory and was selected for use
in the tests here mentioned.
Experiments in making the iodine determinations with stand-
ards of iodine in chloroform indicated that a very satisfactory
color was one having the intensity produced by 1.0 milligram
of iodine dissolved in IO cc. of chloroform. To reproduce such
a tint with Fuchsin S it was found necessary to dissolve 0.02
gram of the dye in 200 cc. of water made acid with 3-5 per cent
of hydrochloric acid and dilute in turn IO cc. of this solution to
200 cc. with water similarly acidified with hydrochloric acid. A
standard prepared in this way and compared in a Duboscq color-
imeter against a solution of I .52 milligrams of iodinedissolved
in IO cc. of chloroform gave a series of readings the average of
which was 14.95 of the Fuchsin S solution equaled IO of the chloro-

form solution, from which it is seen that I 52- = o. 102 per I cc.
14.92
or 1.02 milligram of iodine equivalent to IO cc. of the standard
Fuchsin S solution. Similar readings were made with other
Fuchsin S solutions prepared fresh and allowed to stand various
lengths of time and it was found that the desired tint could be
 Atherton Seidell 393
reproduced at pleasure and remained unaltered for at least three
months, provided the solutions were kept acid. In addition to
this standard corresponding to 1.0 milligram of iodine per IO cc.
of solution it was also found advantageous to prepare a series of
test tubes with gradually increasing intensities of the pink color
corresponding to 0.025, 0.05, 0.075, 0.10, 0.15, 0.20 and 0.25
milligram of iodine per 5 cc., and also in smaller test tubes a
series corresponding to 0.01, 0.02, 0.03 and 0.04 milligram
iodine per I cc. Provided with such standards it is a simple
matter to shake out the liberated iodine from the solutions of
the melt of the samples examined, with I, 5 or IO cc. of chloro-

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form as the amount of iodine might require and compare the
solution so obtained with the corresponding standards.
The several points of advantage, such as saving of chloroform,
permanence and uniformity of standards resulting through the
use of such pink dye solutions as above described will be readily
recognized by anyone making frequent calorimetric determina-
tions of iodine. The dye standards have been in use in this
laboratory with entire satisfaction for several months.
Attention is to be directed also to the applicability of the
above described method to the rapid estimation of iodine in
many pharmaceutical preparations-such as U. S. P. desiccated
thyroid glands, etc.
Finally, it is of interest to note that Dr. J. H. Kastle of the
Hygienic Laboratory has found that the pink colors obtained
with Griess’s reagent for nitrites closely corresponds with the
color of iodine dissolved in chloroform. He has therefore made
use of an acid solution of Fuchsin S as a permanent standard
for nitrites in potable waters with good results.