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Pathlogical Examination of Urine Sample

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11 views27 pages

Pathlogical Examination of Urine Sample

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ansarifaraz1885
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Pathological examination of urine

COMPETENCY NO. – BC 14.4

Dr. Randhir Kumar Pandey


Associate Professor
Department of Biochemistry
PHYSICAL EXAMINATION OF URINE

1. Colour - amber yellow


2. Appearance - clear and transparent
3. Odour - aromatic odour
4. Volume - 1–2 L / day
5. pH - 5 to 8
6. Specific gravity - 1.006-1.024
BIOCHEMICAL EXAMINATION OF URINE

1. Proteins
2. Reducing sugars
3. Ketone bodies
4. Bile salts
5. Bile pigments
6. Blood
BIOCHEMICAL EXAMINATION OF URINE
(1) TEST FOR PROTEINS
Heat Coagulation Test

PRINCIPLE –If proteins are present in urine, due to heating , these are denatured,
and their specific structure gets disrupted. Denatured proteins appear as white
precipitate. Presence of turbidity( which persist after the addition of 3% acetic acid),
indicates the presence of proteins in urine.

NOTE: Calcium, Magnesium, Phosphates are also precipitate due to heating, they are
soluble in acidic medium.
REAGENTS – 3% Acetic Acid

PROCEDURE – Fill 2/3rd of small test tube with protein


solution and add a drop or two of 3% dilute acetic acid.
Mix and hold the test tube at its bottom and heat upper
part of solution gently by rotating the test tube. Formation
of white gelatinous coagulum at upper part of test tube
indicates the presence of heat coagulable protein.
OBSERVATION TABLE –

EXPERIMENT OBSERVATION INFERENCE


Heat coagulation Turbidity seen in upper Protein is present in
part of test tube after urine sample
adding 3% acetic acid
Interpretation
PROTEINS
Presence of proteins in urine is called proteinuria.
 Nephrotic syndrome
 Diabetic nephropathy
 Glomerulonephritis
 Multiple Myeloma
 Urinary Tract Infection
TEST FOR REDUCING SUGARS
BENEDICT’S TEST
PRINCIPLE –In hot alkaline medium reducing sugar gives enediol, enediol being
strong agent reduces cupric ions to cuprous ion forms Cu(OH)2 (Yellow color) ,
which then converted into Cu2O (red color).

The original color of benedicts reagent is blue. It changes to green, yellow. Orange
and brick red.

The test is nonspecific for glucose , some other carbohydrates such as fructose,
galactose, lactose, pentoses and by noncarbohydrates such as ascorbic acid,
salicylates, creatine and uric acid give positive test.
REAGENTS –
1. Sodium carbonate- Provides alkaline medium
2. Sodium citrate- Prevents precipitation of cupric hydroxide
3. Copper sulphate- Provide cupric ions
PROCEDURE –

• Take 5ml of Benedict’s reagent in test tube

• add 8 drops of urine sample

• Mix and then boil for 2 minutes

• Formation of yellow, green, orange or brick red precipitate indicates the

presence of reducing sugars.


OBSERVATION TABLE –
EXPERIMENT OBSERVATIO INFERENCE
N
Benedict’s Test Green /Orange/ Reducing sugar is
Yellow/Brick Red present in urine
precipitate appears. sample.
Interpretation
• The presence of glucose in urine is called Glycosuria.

• The normal renal threshold for glucose is 180 mg/dl.

• When the plasma glucose levels exceeds the renal threshold, the renal tubules cannot reabsorb
all the filtered glucose and causes glycosuria.

• Causes:

• Diabetes Mellitus

• Hyperthyroidism

• Hyperpituitarism

[Physiological glycosuria occurs in Pregnancy due to reduction in renal threshold for glucose]
(3) TEST FOR KETONE BODIES

I. ROTHERA’S TEST
Principle-Nitroprusside used in this test react with both acetone
and acetoacetic acid in the presence of alkali( ammonium hydroxide)
to produce a purple color compound.
Reagents -
Ammonium sulphate
Sodium nitroprusside
Liquor ammonia solution
PROCEDURE –
• Take 5ml of urine in test tube.
• Saturate it with solid ammonium sulphate
• Add to it 2-3 drops of solution of sodium nitroprusside
• Add 2-3ml of liquor ammonia along the inner wall of test tube
• Wait for 5-10 minutes.
• A purple color ring appears between at the junction of two liquids.
• It indicates presence of ketone bodies.
OBSERVATION TABLE –
EXPERIMENT OBSERVATION INFERENCE
Rothera’s Test A purple color ring Presence of Ketone
appears at the junction of bodies in urine sample
two liquids
Interpretation
Ketone bodies: acetoacetate, beta-hydroxybutyrate & acetone.
• Formed in liver from Acetyl Co-A.
• Used in extra hepatic tissue to obtain energy.
• When carbohydrate metabolism is impaired, rate of formation of
ketone bodies increases, the condition is known as ketosis.
• Excretion of ketone bodies in urine known as ketonuria.

• Causes:
• Prolong starvation
• Alcoholism
• Diabetic keto acidosis (Diabetes mellitus)
TEST FOR BILE SALTS
HAY’S SURFACE TENSION TEST

PRINCIPLE – Bile salts when present in urine, lower the surface


tension of urine, When sulfur powder is sprinkled on the surface of
urine, sulfur particles sink to the bottom of the test tube. In the
absence of bile salts, sulfur particles float on the surface of urine.

REAGENTS – Yellow Sulphur Powder


PROCEDURE –
• Take a small test tube and fill 2/3rd with of urine sample.

• Sprinkle some Sulphur powder over the surface of urine sample.

• Do not shake the test tube.

• If Sulphur powder sink, it indicates the presence of bile salt in urine.


OBSERVATION TABLE -
EXPERIMENT OBSERVATION INFERENCE
Hay’s test Sulphur powder Bile salts are
sink in urine sample present in urine.

Interpretation
BILE SALTS
• Hepatocellular jaundice.
• Obstructive jaundice.
TEST FOR BILE PIGMENTS
FOUCHET’S TEST

PRINCIPLE – Bilirubin is precipitated by barium chloride, and it


is oxidized by Fouchet reagent to green biliverdin

REAGENTS – Fouchet reagent

PROCEDURE-Add 5 ml of 10% Bacl2 10 ml of acidified urine,


mix and filter it. Allow the filter paper to dry. A blue or green color
appears at the edges of solution, indicates presence of bilirubin.
OBSERVATION TABLE –

EXPERIMENT OBSERVATION INFERENCE


Faucet's test A blue or green color Bile pigments are
appears at edge of present.
filter paper

Interpretation
BILE Pigments
• Hepatocellular jaundice.
• Obstructive jaundice.
TEST FOR BLOOD
ORTHOTOLUIDINE TEST

PRINCIPLE – This test is more important in excluding the


presence of blood in stains. It is similar to benzidine test.
Orthotoluidine is oxidized by nascent oxygen to form a blue or
green colour compound which is unstable.

REAGENTS – Orthotoluidine Solution


Hydrogen Peroxide (H₂O₂)
PROCEDURE –
• Take 3ml of urine sample in a test tube.
• Add 1ml of Orth toluidine solution
• add 1ml of H₂O₂.
• Mix it.
• Development of blue or green colour
indicates the presence of blood in given
fluid.
OBSERVATION TABLE -
EXPERIMENT OBSERVATION INFERENCE
Orth toluidine test Transient Blue colour is Presence of blood in
obtained. sample.

Interpretation

• Presence of intact RBC in urine is known as Hematuria.


• Presence of free hemoglobin in urine is known as Hemoglobinuria.
Causes of Blood in urine:
 Cancer of urinary tract,
 Stone in urinary tract
 Blood disorders such as hemophilia, leukemia .
 Typhoid fever
 Extensive burns
 Mismatched blood transfusions
THANK YOU

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