JULY 24, 1954THE CHILD WITH HEART DISEASE
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A NEW TABLET TEST FOR BILIRUBIN
J. A. TALLACK, M.B., Ch.B.
SHEILA SHERLOCK, M.D., F.R.C.P.
Physician and Lecturer
Department of Medicine, Postgraduate Medical School of
This paper describes a simple method for the detection
of small amounts of bilirubin in the urine.
first concentrated by adsorption on a special test mat
and then allowed to react with a stable diazonium com-
pound incorporated in a tablet.
urine from normal and jaundiced subjects have been
compared with those using two standard routine methods
for urinary bilirubin, Fouchet's test and the iodine ring
The tablet test is performed as follows: (1) Place 5 drops
of urine on the test mat, which is 19 mm. square and
2 mm. deep, and is composed of a mixture of asbestos
and cellulose fibres.
of the moistened area of the mat.
dye (p-nitrobenzene diazonium p-toluene sulphonate) and
also sulphosalicylic acid, sodium bicarbonate, and boric
(4) The colour developing on the mat is recorded
within 30 seconds.
The bilirubin is concentrated on the test
The diazo dye is taken into solution by the water,
aided by the effervescence caused by the presence of sul-
phosalicylic acid and sodium bicarbonate in the tablet;
the boric acid is a vehicle.
urine the mat around the tablet turns purple and the amount
of bilirubin is roughly proportional to the speed of develop-
ment and intensity of the colour.
after 30 seconds is ignored.
satisfactory colour reaction
water is allowed to remain on the tablet for a few seconds
before the second drop is put on to wash the fluid on to
the test mat.
Fouchet's test is performed by adding 2 ml. of 10%
barium chloride to 10 ml. of urine, filtering and adding
Fouchet's reagent to the filter paper.
indicated by a green colour.
The iodine ring test is performed by layering 2 ml. of
tincture of iodine diluted
of urine in a test-tube.
A positive result
by a green ring at the interface.
A modification of the tablet test has been employed, using
5 drops of urine on the test mat as before, but replacing
the tablet with
result is indicated by the development of a green colour
on the mat.
Results obtained with
(2) The tablet is placed on the centre
It contains a stable diazo
(3) Two drops of water are allowed to flow on the
If bilirubin is present in the
Any colour developing
It is found that the most
is given if the first drop of
A positive result is
1 in 4 with water on to 10 ml.
1 drop of Fouchet's reagent.A positive
Quantitative urinary bilirubin estimations were performned
by the method of Golden and Snavely (1948).
Schlesinger's test with alcoholic zinc acetate and by Ehrlich's
Serum bilirubin levels were estimated by
the method of King and Coxon (1950).
Samples of urine from 100 non-jaundiced patients picked
at random from the wards of a general hospital and 200
urines from jaundiced patients were tested.
urines containing urobilin or an excess of urobilinogen but
no bilirubin were also studied.
bilirubin level was estimated at the time of testing the
The 100 urines from non-jaundiced patients were tested
to determine whether false-positive reactions might result
from the administration of drugs or from the presence of
other abnormal substances in the urine.
receiving many varied forms of treatment and suffering from
a wide range of disorders.
give a positive tablet reaction, and it therefore seems unlikely
that false-positive reactions occur owing to the presence of
commonly administered drugs in the urine.
there was seen on the mat a faint orange-brown colour
which became more conspicuous after the mat had been
standing for some time.
This colour did not interfere
with the reading of the test, and was presumably due to
in the urine of diazotizable non-bilirubin
The presence of urobilin or an excess of urobilinogen
does not give
36 urines with a positive Ehrlich or Schlesinger reaction
were negative both by the tablet test and by Fouchet's
These results agree with those previously reported with
the tablet test.
Free and Free (1953) tested approximately
2,000 urine samples from general-hospital patients and failed
to find any substance giving false-positive reactions.
also tested 100 urine samples from normally healthy adults;
99 gave negative reactions and one an unexplained trace
reaction. Klatskin and Bungards (1953) found two false-
positive tablet reactions in a series of 478 urines, and in both
these instances there might have been some hepatic dysfunc-
Giordano and Winstead (1953) found that the pre-
sence of increased amounts of urobilinogen in the urine
did not mask the colour reaction as it did in the Harrison
spot test, in which Fouchet's reagent is used.
Of the 200 urines tested which gave either a positive
Fouchet or a positive tablet reaction, 185 were positive for
both methods; 192 gave a positive Fouchet and 193 a
positive tablet reaction, while only 135 were positive with
the iodine method.
Ninety urines giving a positive tablet reaction were also
tested with the modification using the test mat and replacing
the tablet by Fouchet's reagent: 58 gave a positive result
and 32 a negative one.
These results suggest that the tablet test and Fouchet's
method are of about equal sensitivity and are considerably
superior to the iodine ring test and to the modification
for the detection of bilirubin in the urine.
found that the tablet test was more sensitive than methods
using Fouchet's reagent (Klatskin and Bungards,
Giordano and Winstead, 1953).
Quantitative analysis of bilirubin in urine is made difficult
by the presence of interfering substances.
attempt was made to compare the quantitative urinary
bilirubin values with the results for the qualitative tests.
Eighteen urines were analysed and then diluted serially, and
the lowest concentration giving a positive qualitative reac-
tion was recorded (Table I).
the tablet test could usually detect bilirubin between 0.1
In 92 instances the serum
The patients were
In no instance did the urine
Altho>ugh -the results varied,