TEST FOR ABNORMAL

CONSTITUENTS OF URINE
BI 11.4 & 11.2
EXPERIMENT OBSERVATION INFERENCE

1.BENEDICT’S REDUCTION
TEST-

5ml of Benedict’s reagent is Brick red precipitate is Reducing sugar is present

taken in a test tube, to it 8 formed. in the given sample.
drops of urine sample is
added & mixed and boiled for
2 minutes.
1.BENEDICT’S TEST-
Principle- Reducing sugar in hot alkaline
medium are converted into ENEDIOL form
which are strong reducing agent. They reduces
the cupric ion in benedict’s solution to cuprous
ions which gives red precipitate of Cu₂O .
• Benedict’s test is qualitative as well as
semi-quantitative test.

A precipitate of either green, yellow, red

colour indicates the presence of reducing
sugar and is depends upon concentration of
sugar in urine and therefore a rough
estimation of sugar in urine can be made in
the following way.
 COLOUR SUGAR CONTENT

Green colour / Light upto 0.5 g%

green precipitate
Green precipitate 0.5 to 1 g%(+)

Yellow precipitate 1 to 1.5 g%(2+)

Orange precipitate 1.5 to 2 g%(3+)

Red precipitate >2 g%(4+)

• Q. Name some condition in which benedict’s
test is positive?

• Q. Name non-carbohydrate compounds which

also gives positive benedict’s test?

• Q. What do you mean by “renal threshold”?

• Q. What is glycosuria? Enumerate the causes

of glycosuria?
EXPERIMENT EXPERIMENT INFERENCE

2. MODIFIED BOILING TEST

FOR PROTEIN-

3/4th of test tube is filled Turbidity appeared in

with urine sample. Upper heated portion of urine. Protein is present in given
one third of test tube is sample.
heated.

1-2 drops of 33% of acetic Turbidity did not dissolved.

acid is added.
Modified boiling test-

Principle- On boiling heat coagulable protein

coagulates which appears as turbidity.

Q. What do you understand by the term

“proteinuria”?

Q. Causes of proteinuria?
EXPERIMENT OBSERVATION INFERENCE

3. ROTHERA’S TEST FOR

KETONE BODIES

5 ml of given urine sample is

taken in a test tube and Permanganate colour ring Ketone bodies present
saturated with ammonium is observed at the junction in the given urine
sulphate. 2 drops of freshly of two liquid. sample.
prepared solution of sodium
nitroprusside is added and
mixed. 2 ml of ammonia
(ammonium hydroxide) is added
by the side wall of test tube.
• ROTHERA’S TEST-

Principle- Ketone bodies forms a complex with sodium

nitroprusside in alkaline medium developing a permanganate

colour.

Q. Name some condition in which ketone bodies are present in

urine?

Q. Which ketone body does not give positive rothera’s test &

why?
EXPERIMENT OBSERVATION INFERENCE

4. HAY’S SULPHUR TEST FOR

BILE SALTS-

Two test tube is taken. In first

test tube 2/3rd is filled with Sulphur powder sinks Given urine sample
urine sample and second test in the test tube contains bile salts.
tube is filled with water. Pinch containing urine
of sulphur powder is sprinkled sample.
in both test tube.
• HAY’S SULPHUR TEST
Principle- Bile salts reduces the surface tension of
urine due to which sulphur powder sinks.
Q. Name bile acids and bile salts?
Q. Bile salts are formed from which compound?
Q. Functions of bile salts in body?
Q. Condition in which this test is positive?
EXPERIMENT OBSERVATION INFERENCE

5. FOUCHET’S TEST FOR BILE

PIGMENT-

15-20ml of urine sample is taken

in a test tube, to it 1-2 drops of Greenish blue colour Bile pigment is present
saturated magnesium sulphate appeared at the place in given urine sample.
solution and 5 ml of 10% BaCl2 where fouchet’s reagent
is added and mixed. Let the test is added.
tube stands for 10 minutes. It is
then filtered with filter paper.
Spread the filter paper over a
watch glass and one drop of
Fouchet’s reagent is added to
one part of precipitate on filter
paper.
• FOUCHET’S TEST-
Principle- Barium chloride reacts with sulphates
present in urine to form barium sulphate. It
adsorbs bilirubin present in urine which is
precipitated. Bilirubin is oxidised to green biliverdin
by fouchet’s reagent(FeCl₃).

Q. Name bile pigments?

EXPERIMENT OBSERVATION INFERENCE
6. BENZIDINE TEST FOR BLOOD
PIGMENTS-

2ml of urine is boiled and

cooled in one test tube.

In another test tube knife point

of benzidine powder is
dissolved with 2 ml of glacial
acetic acid then 2 ml of Deep blue colour was Blood pigment is
hydrogen peroxide is added observed in the test tube present in given urine
and mixed. containing urine sample. sample.
Then it is divided in two parts.
To first part 5-10 drops of
boiled and cooled urine sample
is added.
To second part 5-10 drops of
water is added
• BENZIDINE TEST-
Principle- The peroxidase present in RBC splits
H₂O₂ to produce nascent oxygen which reacts
with benzidine in acidic medium to produce blue or
green colored complex.

Q. What is haematuria? Enumerate its causes?

THANK
YOU