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BILE PIGMENTS. 801
and then with a solution of sodium or ammonium carbonate. The pre-
cipitate which contains the bile-coloring matter is filtered, washed, dis-
solved in alcohol which contains 5 cc. of concentrated hydrochloric acid
in 100 cc. (I. Munk), and heated to boiling, when the solution becomes
green or bluish green. According to Xakayama i
this reaction is more
delicate on using a mixture of ferric chloride, acid, and alcohol.
Hammarsten’s Reaction. For ordinary cases it is sufficient to add
a few drops of urine to about 2-3 cc. of the reagent (see page 432), when
the mixture immediately after shaking turns a beautiful green or bluish
green, which color remains for several days. In the presence of only
very small quantities of bile-pigments, especially when blood or other
pigments are simultaneously present, pour about 10 cc. of the acid or
nearly neutral (not alkaline) urine into the tube of a small centrifugal
machine and add BaCb solution and centrifuge for about one minute.
The liquid is decanted and the sediment stirred with about 1 cc. of the
reagent and centrifuged again. A beautiful green solution is obtained
which may be changed, by the addition of increased quantities of the acid
mixture, to blue, violet, red, and reddish yellow. The green color may
be obtained in the presence of 1 part bile-pigment in 500,000-1,000,000
parts urine. In the presence of large amounts of other pigments calcium
chloride is better suited than barium chloride.
Bouma 2
has suggested the use of alcohol containing ferric chloride
and hydrochloric acid instead of the above-mentioned acid mixture. He
has also wr
orked out a colorimetric method of quantitative estimation
of bilirubin in urine by means of this reagent.
As above indicated, we have a great many other tests besides these
given above. A very complete summary of these tests and the literature
thereof can be found in the work of Obermayer and Popper.
For ordinary purposes the above-mentioned tests are sufficiently
delicate, and according to Hammarsten it is not advisable, as also in the
case of the detection of proteid, sugar, etc., to increase the delicacy of
a test so that it shows the presence of the traces of the questionable
substance in normal urine. If in certain cases a greater delicacy is
required than is obtained with the above tests, then we must recommend
the flotation test of Obermeyer and Popper 3
with iodine and salt.
Medicinal coloring-matters produced from santonin, rhubarb, senna, etc.,
may give an abnormal color to the urine and may be mistaken for bile-pigments,
or, in alkaline urines, perhaps for blood-eoloring matters. If hydrochloric acid
is added to such a urine, it becomes yellow or pale yellow, while on the addition
of an excess of alkali it takes on a more or less beautiful red color.
1
Munk, Arch. f. (Anat. u.) Physiol., 1898; Nakayama, Zeitschr. f. physiol. Chem.,
36.
* Deutsch. med. Wochenschr., 1902 and 1904
* Wien. med. Wochenschr., 21.
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