TABULATION FOR CHEMICAL TESTS FOR URINE

TESTS FOR BILIRUBIN

TEST/METHOD

PRINCIPLE

Smiths Test (Iodine

test)
Gmelins Test

Ultzmans Test

After precipitation of the bile pigments by HNO3, it is

oxidized by acids to derivitives such as biliverdin (green),
bilicyanine (blue) and choletelin (yellow).

REAGENTS

RESULTS

*2mL of 0.7% iodine in 95% ethyl

alcohol

Positive: Emerald green color at the zone of

contact.

*Concentrated nitric acid (with

yellow nitrous acid)

Positive: Band of different colors (green,

violet, yellow, red, blue-green) between the
zones of contact.

*25%
potassium
solution,

hydroxide
Postive: A definite color green

*pure hydrochloric acid

Positive: Foam has a yellow color that
persists for several minutes.
Foams Test

*Ictotest tablet
Ictotest Tablet Test
*H2O

NOTE: Drugs such as pyridium, serenium,

acriflavin, and urobilin compound also
gives yellow foam.
Positive: Bluish-purple color develops
around the tablet on the mat.
NOTE: The color on the mat around the
tablet should be observed exactly 30
seconds later.

Nitric Acid Test

Harrison Spot Test

Methylene Blue Test

Concentrated nitric acid

After precipitation of the bile pigments by barium

chloride (BaC12), it is oxidized by acids to derivatives
such as biliverdin (green), bilicyanin (blue) and choletelin
(yellow).

*10% Barium chloride

Positive: A display of colors, (yellow,

green and blue).
Positive: A blue to green color.

*Fouchets reagent (trichloracetic acid

25 gm., FeCl3 solution 10% - 10 cc.,
and distilled water100 cc.)
*0.2% aqueous solution (methylene
blue drop by drop to 5 cc., of a pre
breakfast urine)

Positive: If more than 4 drops was

added when the blue color appeared.

* methylene blue

Negative: If 4 drops or less has been

added when the blue color appeared.

Rosenbach Test

*Nitric acid containing yellow nitrous

acid

NOTE:
*Multiply the number of drops by the
dilution if more than 8 drops is required.
Make a 1-5 dilution of urine and repeat
the test.
* False positive result may be obtained
with urine having high SG and after
large doses of penicillin, riboflavin or
quinacrine (atabrine).
Positive: A band of colored rings
around the nitric acid.

Maher Test

*20% solution of paratoluene sulfonic

acid (10% HCL may also be used)
*0.1% freshly prepared
sodium nitrate

solution of

Positive: Green color

Huppert Test

Ferric Chloride
Test

The bile pigments are precipitated as compound with

calcium hydroxide. The compound is dissolve and the bile
pigments are oxidized to a green-colored compound in an
acid alcohol solution.

The bilirubin in the urine is concentrated by absorption on

barium sulfate and phosphate and then is oxidized by
ferric chloride in acid solution to biliverdin (green) and
cholecyanin (blue)

*Milk of lime (suspended 15 gm. Of

reagent CaO in 1 liter of water)
*Concentrated HCL,
*Ethyl alcohol, 95%

*10% Barium chloride

Positive: Green or blue color
*Fouchets reagent

*Thick filter paper (Schleicher and

Schuell, No. 470) that has been soaked
in saturated barium chloride, dried and
cut into small strips,

Oxidation Method

*Fouchets reagent

Fouchets Test

Barium chloride is added to te urine. A precipitate of

barium sulfate is produced on to which the bilirubin is
absorbed. The barium sulfate absorbed bilirubin is filtered
off and a drop of Fouchets ferric chloride solution added
to the precipitate. The ferric chloride oxidizes the bilirubin
to biliverdin, producing a greenish-blue spot.

Positive: Green color

Postive: A green color usually

deepening on standing because the
formation
of
biliverdin.
NOTE: High levels of urobilin or
indicant may cause a brown-purple
color; salicylates give purple color; high
levels of urobilinogen may cause red
color of the green of biliverdin.

*10% barium chloride

Positive: Greenish-Blue color

*Whatman No. 1 filter

NOTE: Filtrate from this test may be

used for the Schlesingers test for
urobilin.

*Fouchets reagent

Positive: Green color

*Tablets of plastic of Paris
Franklin Test
*Fouchets reagent

NOTE: Fouchets reagent gives a purple

color with salicylates nitric acid does
not do so. Bile gives a green color with
Obemayer reagent (color not soluble in
chloroform).

BiliLabstix

TESTS FOR CATHECHOLAMINES

TEST/METHOD
Fluorometric Test

PRINCIPLE

REAGENTS

RESULTS