URINE ANALYSIS

Normal constituents of URINE

Water -90-95% Solid-5-10%%

Inorganic Organic

Chlorides=10-15g/d Urea=25-30g/d
Sodium=3-5g/d UA= 05.-0.8g/d
Potassium=2-2.5g/d Creatinine=1-18g/d
Calcium=0.1-0.3g/d Ammonia=0.7-0.8g/d
Phosphates=0.8-1.3g/d
Sulphates=1.0-1.2g/d
 URINE ANALYSIS

Microscopic Biochemical
Bacteriological Analysis
analysis analysis

Routine
Qualitative Quantitative

Physical Chemical
 Types Of Urine Specimen
• 1) Mid stream urine sample
• 2) 24-hour urine sample
• 3) Timed urine sample
• 4) Spot urine sample

Preservatives:
• Thymol
• HCl
• Refrigeration
 AIM
To Analyse urine for its
Abnormal Constituent
 Physical Analysis

• 1) Colour
• 2) Odour
• 3) pH
• 4) Specific Gravity
• 5) Volume
 COLOUR

Normal Straw Coloured

Yellow in Jaundice Red in Haematuria Brown in Hburia

Cloudy in infection Black to brown in alkaptonuria

 ODOUR
Normal- ammoniacal odour.

Pathological
1) Fruity odour in diabetic ketoacidosis
2) Burnt Sugar in maple syrup urine disease.
3) Putrid odour in bacterial infection
 VOLUME
• Normal : 800-2500ml/24hr; average=1500ml/24hr

• Polyuria: urine output >2500ml/d

DM DI Diuretics

• Oliguria: Urine output <400ml/d

Nephritis Diarrhoea Excessive
vomiting

• Anuria: Urine output< 100ml/d

Shock Acute renal failure
Incompatible BT
 SPECIFIC GRAVITY
• Normal: 1.016-1.025; Average=1.020
• Instrument used to measure specific gravity=
urinometer

Specific gravity
• DM
• Dehydration

Specific gravity
• Diabetes insipidus
• Excessive water intake
 pH
Normal= 4.5-8; average=6.0

Urinary pH<4.5
• Acidosis
• Ketoacidosis

Urinary pH>8
• Alkali therapy
• UTI
 CHEMICAL ANALYSIS

Routinely analysed abnormal constituents of

urine are:
• 1) Reducing Sugar
• 2) Proteins
• 3) Ketone bodies
• 4) Bile Pigments
• 5) Bile salts
• 6) Blood
 1. TEST FOR REDUCING SUGARS
BENEDICT’S TEST
• PRINCIPLE: Reducing sugars under alkaline
condition form enediols which are powerful reducing
agents. They reduce cupric ions (Cu2+) to cuprous
form (Cu+) and form the cuprous oxide (Cu₂O)ppt
(red).
BENEDICTS REAGENT

Copper Furnishes cupric ions (Cu++)

sulphate

Sodium Makes medium alkaline

carbonate

Sodium Complexes with the Cu⁺⁺ ions so

citrate that they do not deteriorate to Cu⁺
ions during storage
PROCEDURE:
• Pipette 5 ml of Benedict’s reagent in a test tube.
• Add 8 drops of urine to the Benedict’s reagent.
• Heat carefully on a flame of a gas burner.
Colour Sugar
OBSEVATION:
Green 0.5%-1%

Yellow 1.0-1.5%

Orange 1.5-2%

Bric red >2%

ppt
Reducing sugars: Have free keto or aldehyde group or
anomeric carbon. Example: Monosaccharide and
Disaccharides with free functional group.

Reducing disaccharide

Maltose Glucose + Glucose α1→4 glycosidic bond

Isomaltose Glucose + Glucose Α1→6 glycosidic bond

Lactose Glucose + Galactose β1→4 glycosidic bond

Non-Reducing disaccharide
Sucrose Glucose + Fructose α1→β2 glycosidic bond
BENEDICT’S TEST:
• POSITIVE:
1) Glucosuria: DM 2) Lactosuria 3) Galactosuria
4) Pentosuria 5) Fructosuria

• FALSE POSITIVE:
1) Ascorbic acid 2) Uric acid 3) Drugs: Salicylates

• Non-specific test.
• Semiquantitative test.
• Not given by Sucrose.
 2. TEST FOR PROTEINS
• a) Heat Coagulation Test
• PRINCIPLE: Protein is denatured on heating; denatured
protein is less soluble than the native protein and forms a
white coagulum.
 Procedure Observation

Fill 3/4th of test tube with urine White coagulum observed

sample. Heat upper 1/3rd of tube in the heated portion
taking lower 2/3rd as control.

Add 2-3 drops of acetic acid 1% Coagulum gets

intensified/persists
• In alkaline medium heating ppt phosphates and
carbonates.
• Acidification is required to rule out the ppt
because of them.
• It can be used as semiquantitative test
• b) HELLER TEST: Concentrated acid (HNO₃)
causes denaturation of protein forms a white ring at
the junction of two liquids.

PROCEDURE:
• Take 3 ml of concentrated nitric acid.
• Incline the tube and add carefully 3 ml of urine, so that it
forms the upper layer without disturbing the lower HNO₃
layer.
• In a positive reaction, a white zone of precipitate protein will
appear at the junction of two liquids.
c) SULPHOSALICYLIC ACID TEST

PROCEDURE
• Place 3ml of clean urine in test tube
• From the side of tube add 2-3 drops of
sulphosalicylic acid on top.
• Observe the turbidity.
• Persistence of turbidity even after heating is
indicative of protein.
• Normal urine total protein < 150mg/24hr.
• Proteinuria > 150mg/24hr.

• Normal urine albumin <30mg/24 hr.

• Albuminuria = 30-300mg/24hr.
 COMMON CAUSES OF PROTEINURIA

Pre-Renal Renal Post Renal

Strenuous exercise Diabetic Urinary tract

nephropathy infection

Severe dehydration Nephrotic

syndrome
 TEST FOR KETONE BODIES
Rothera’s Nitroprusside Test

• PRINCIPLE: Sod. Nitroprusside in alkaline medium

reacts with keto group of acetone and acetoacetate
and forms a purple ring.

• PROCEDURE: To 5ml of urine and saturate it with

(NH₄)₂SO₄. (to ppt the protein to and interference)

• Add 2-3 drops of freshly prepared Sod. Nitroprusside

solution and mix well.
• Add 1ml of liquor ammonia along the sides of tube
without mixing.

• OBSERVATION: Purple ring at the junction of two

liquid.
• Normally ketone excretion in urine 1mg/24hr.
• Acetone, acetoacetate and hydroxybutyrate are
known as ketone bodies.
• Test is negative for hydroxybutyrate as it lacks
the keto group.

CAUSES OF KETONURIA
• Uncontrolled DM.
• Starvation.
 TEST FOR BILE SALTS
Hay’s Sulphur Test

• PRINCIPLE: Bile salts lower the surface tension of

solution in which they are dissolved resulting in
sinking of sulphur powder.

• PROCEDURE: Fill 2/3rd of test tube with urine and

gently sprinkle the sulphur powder on it. Run a control
similarly taking DW as control.

• OBSERVATION: In presence of bile salt sulphur

powder sinks to the bottom.
• OBSERVATION: In presence of bile salt sulphur
powder sinks to the bottom.

• Normally bile salt are absent in urine.

• Positive Hay’s test: Obstructive jaundice.

• False positive test: Contamination of urine with

the detergent used in cleaning of tubes.
 TEST FOR BILE PIGMENT
FOUCHET TEST
PRINCIPLE: BaCl₂ reacts with SO₄ in urine to form
BaSO₄; bilirubin gets adsorbed on it and Fouchet’s
reagent oxidizes bilirubin (yellow) into biliverdin
(green).

FOUCHET REAGENT
1) 10% FeCl₂
2) TCA
PROCEDURE: Add 5ml of 10% BaCl₂ to 10ml of
urine and filter. Dry the filter paper and add few
drops of fouchet’s reagent. Note the colour
change.

OBSERVATION: Colour of filtrate changes from

yellow to the green in presence of bilirubin.
CLINICAL SIGNIFICANCE:
• Normally bile pigment in urine is absent or present
in negligible amount.

• Presence is indicative of increased conjugated

bilirubin
➢ Obstructive Jaundice
➢ Hepatitis

• Unconjugated bilirubin is bound to albumin and is

not excreted in urine.
 TEST FOR BLOOD
Benzidine Test

• PRINCIPLE: Haem of Hb decomposes H₂O₂ to O₂

which oxidise the benzidine to blue or green
coloured products.

• PROCEDURE: To 3 ml of benzidine solution add

2ml of urine and 1ml of 3% H₂O₂. Mix and note
the colour.

• OBSERVATION: Blue or green colour immediately.

CLINICAL SIGNIFICANCE
• HAEMATURIA: Presence of blood in urine;
microscopic and macroscopic.
• Causes of Haematuria: Renal stone; trauma;
TB and glomerulonephritis.

CAUTION: Benzidine is carcinogenic, should be

handled carefully.