QUALITATIVE ANALYSIS

OF ABNORMAL
CONSTITUENTS OF URINE
 Physical properties of urine
• Volume
• Color
• Odor
• Specific gravity
• pH
 Volume
• Normal adult urine volume is 1.2 to 1.5 L/day

• Polyuria: Urine output > 3L/day. It is seen in diabetes mellitus, diabetes

Insipidus, end-stage renal disease etc.

• Oliguria: Urine output < 500ml/day. It is seen in condition associated with

dehydration.

• Anuria: Complete absence of urine formation usually or < 50ml/day. It is seen

in acute renal failure.

• Nocturia: Excess excretion of urine at night. It is seen in patients with prostatic

enlargement, diabetes etc.
 Colour
• Normal color of urine is amber, caused by urochrome pigment.

• Dark colored urine is seen in condition associated with dehydration like

diarrhea, dysentery etc.

• Dark yellow colored urine is seen in obstructive jaundice.

• Red colored urine is seen in condition associated with hemoglobinuria,

hematuria etc.

• Black colored urine is seen in alkaptonuria.

• Reddish or brown colored urine is seen in porphyria

 Odor
• Normal - Faintly aromatic odor.

• Long standing urine without any added preservative gives ammoniacal

odor, due to bacterial action which converts urea to ammonia.

• Fruity odor - Diabetic ketoacidosis

• Specific odor :

i. Mousy odor is associated with Phenylketonuria

ii. Burnt sugar odor is associated with Maple syrup urine disease etc.
 Specific gravity
• Normal specific gravity of urine sample ranges from 1.005 - 1.030.

• Minimum specific gravity after a standard water load should be > 1.003 in normal
individuals.

• Increased in diabetes mellitus, dehydration, and Syndrome of Inappropriate ADH

secretion (SIADH)

• Decreased in diabetes insipidus, acute tubular necrosis, and psychogenic polydipsia.

• Fixed specific gravity of 1.010 seen in chronic renal failure.

• Long's coefficient refers to 'total solids excreted in the urine'. The solid content of
1000ml of urine is calculated by multiplying the last 2 digits of specific gravity at 25°C
by 2.6 and is expressed in gm/L.)
 pH
• Normal pH range of urine is 4.8 - 8.
• Pure vegetarian diet increases alkalinity of urine.
• High protein diet decreases pH of the urine.
 AIM

To analyze the abnormal constituents present in the given

urine sample
 TEST FOR PROTEINURIA
PROCEDURE OBSERVATION INFERENCE
HEAT COAGULATION TEST:
Take 3/4th of the test tube with
the given urine sample. Heat
the upper portion of the test
tube by holding the bottom of A white coagulum is formed in
Presence of protein
the test tube using the test the upper part of the test tube
tube holder. Coagulation takes
place. The lower portion serves
as control. Then, add 2 drops of
1% acetic acids.

PRINCIPLE:
It indicates the presence of heat coagulable protein such as albumin. The appearance of coagulation
indicates the denaturation of proteins and addition of acetic acid precipitates protein.
 TEST FOR REDUCING SUGARS
PROCEDURE OBSERVATION INFERENCE

BENEDICT’S TEST: Green or yellow or orange or

Take 5 ml of Benedict’s reagent. brick red color/ precipitate
Add 8 drops of urine sample depending upon the Presence of reducing sugars
and mix well. Heat the test tube concentration of the reducing
for 2 minutes. sugar present in the sample

PRINCIPLE:
Copper sulphate is reduced by the reducing sugar to form red cuprous oxide. The colour formed depends upon the
concentration of the reducing sugar
 TEST FOR KETONE BODIES
PROCEDURE OBSERVATION INFERENCE

ROTHERA’S TEST:
Take 5 ml of urine sample. Add
solid ammonium sulphate till it
get saturates. Add 4 drops of Purple colour ring is formed at Presence of ketone bodies
5% sodium nitroprusside mix the junction of the two layers.
well. Add 2 ml of liquid
ammonia along the sides of the
test tube.

PRINCIPLE:
Ketone bodies reacts with sodium nitroprusside in the presence of alkali to give purple coloured ring.
The test is answered only by acetone and acetoacetic acid
 TEST FOR BILE PIGMENT
PROCEDURE OBSERVATION INFERENCE
FOUCHET’S TEST:
Take 3 ml of urine sample in a test
tube, add 2ml of 10% Barium
chloride (BaCl2) and few crystals of A green precipitate is obtained Presence of bile pigment
magnesium sulfate. Wait for 5 in the filter paper
minutes for the formation of
precipitate. Filter the contents of the
tube using filter paper and discard
the filtrate. Unfold the filter paper
and blot the excess fluid. Add 1 drop
of Fouchet's reagent over the
precipitate in the filter paper.

Principle:
Barium sulphate is formed as precipitate, which adsorbs bile pigments. Ferric chloride in Fouchet's
reagent oxidizes Bilirubin to green Biliverdin.
 TEST FOR BILE SALTS
PROCEDURE
HAY'S SULPHUR TEST:
Take 2 test tubes and label
them as "T" for test and "C" for
control. Take 2ml urine sample
in "T tube and distilled water in
"C" tube. Sprinkle Sulphur
powder over the surface in
both the test tubes.
OBSERVATION INFERENCE
Sulphur powder sinks to the Presence of bile slats
bottom of the test tube
containing bile salts in urine. (T)
Sulphur powder floats in (C) test
tube.

Principle:
Bile salt present in urine lowers the surface tension of urine and so sulphur powder
sinks.
 TEST FOR BLOOD
PROCEDURE OBSERVATION INFERENCE

BENZIDINE TEST: A transient blue or green colour Presence of blood in urine

Take a clean dry test tube, add is obtained
a pinch of Benzidine powder,
1ml Glacial Acetic acid, 1 ml
hydrogen peroxide followed by
1ml of urine sample

PRINCIPLE:
Hb decomposes hydrogen peroxide releasing nascent oxygen which oxidizes Benzidine reagent to
form the characteristic colour.
 RESULT

The given urine sample contains protein, reducing sugars, ketone

bodies, bile pigment, bile salts and blood.
Thank you