URINE EXAMINATION

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 Composition
Water – 95%
2% - urea
3% organic & inorganic substances

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 Sample collection
 Wide mouthed glass bottles
 Disposable containers / polyethylene bags (infants)
 Freshly voided, clean catch, mid stream specimen of urine
 Types of samples – first morning (ideal, concentrated)
random specimen – chemical screening
2nd voided sample – glucose
 Timed specimen – 24 hr (glucose, protein, hormones)

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 Preservation / storage
 Ideally examined within 1 – 2 hrs of voiding
 24 hr specimen/ distant labs – preservatives
 Changes during preservation – pH, turbid, bacterial
contamination, lysis of RBCs
 Ideal preservatives – no chemical alteration, preserves the
sediment
 Toluene – best for chemical examination
 Formalin – preserves sediment
 Urine culture - preservatives are not added

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 PHYSICAL EXAMINATION
•
Volume
• Normal – 600 to 2000 ml / day ; more during day
• Polyuria - > 2000 ml in 24hrs
• Causes - Physiological, cold weather
high intake of fluids
Diabetes mellitus, Diabetes insipidus.
• Oliguria - < 500 ml in 24hrs

• Causes – hot weather

decreased fluid intake
renal / cardiac diseases

• Anuria – complete suppression of urine formation < 50 ml /day

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 COLOUR

• Normal – pale yellow to dark amber due to urobilinogen

• Dilute urine – pale yellow ( DM, DI)

• Concentrated urine – deep yellow

• Yellow - bilirubin

• Milky urine – purulent ( UTI ), chyluria

• Orange – increased urobilinogen

• Red – hematuria, beet root ingestion

• Black – hemoglobinuria , Alkaptonuria

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 APPEARANCE
• Normal – clear
• Cloudy –
– amorphous phosphates & urates – heat disappear
– Pus, bacteria – centrifuge – sediment
– fat & chyle – centrifuge – supernatant

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 ODOUR
• Normal – No smell or slight aromatic odour (allowed to stand

- ammonical smell)

• Fruity odour – ketone bodies

• Pungent smell – bacterial contamination

• Mousy odour – phenyl ketonuria

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 REACTION & pH

• Normal – slight acidic pH – 4.6 to 7(6.0) due to presence of

weak acids ,phosphates
• Acidic urine –
– respiratory acidosis(  CO2)
– metabolic acidosis (DM, Starvation)
– high protein intake, E.coli infection

• Alkaline urine –
– respiratory alkalosis (hyperventilation)
– metabolic alkalosis (vomiting)
– Proteus, Pseudomonas infection
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 SPECIFIC GRAVITY

• Amount of solids in solution of urine

• Used to measure the concentration power of kidneys
• Normal – 1.015 to 1.025 in a 24 hr specimen
• Substances influencing – urea, NaCl, PO4, albumin, sugar
• Hyperesthenuria – high specific gravity (DM, dehydration)
• Isosthenuria – fixed S.G of 1.010,poor tubular absorption
• Hyposthenuria – S.G < 1.007(pyelonephritis,DI,alcohol,
diuretics)

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 DETERMINATION OF SPECIFIC GRAVITY

Urinometer
• Readings from 1.000 to 1.060 with divisions of 0.001 to 0.002
• Corrections for temperature, albumin, other solids
• Calibrated at 15o
• Every 3o rise or fall add/subtract 0.001
• Urine strip method
– Refractometer - based on refractive index of solids

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 CHEMICAL EXAMINATION
PROTEIN
Qualitative test – Heat & Acetic acid test
Principle : coagulation of proteins by heat
Procedure:
• urine made acidic by adding a few drops of acetic acid
• Filter turbid urine
• Take a long column of urine in a test tube.
• Boil the upper half( lower half acts as a control for
comparison)

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 White coloured cloud in upper half

3 % acetic acid

Disappears persists

Phosphates , carbonates Albumin

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BENCE JONES PROTEIN

White precipitate appears when hot (40 to 60° c) and disappears

on boiling & reappears on cooling

Other tests

• 3 % Sulphosalicylic acid test : 2ml SSA + 2ml urine – turbidity

• conc. HNO3 test

• Urine dipstick

• Normal amount of proteins in the urine – 30 to 50 mg / 24 hrs

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 PROTEINURIA

• Massive Proteinuria - > 3.5 gms / day

Nephrotic syndrome, Glomerulonephritis, CCF
• Moderate Proteinuria 0.5 to 3.5 gms / day
Chronic GN, Diabetic nephropathy, multiple myeloma, renal
stones
• Minimal Proteinuria < 0.5 gms/ day
Polycystic kidney, orthostatic proteinuria
• Microalbuminuria 30 – 300mg/ day
Indication of future renal failure in DM, HT

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 SUGAR

Benedict’s test – semi quantitative test

Principle : sugar reduces copper sulphate in an alkaline solution into

insoluble cuprous oxide

Benedict’s reagent – CuSO4 + Sod. Carbonate, Sodium citrate

Procedure :

1. Take 5 ml of Benedict’s reagent in a test tube

2. Add 8 drops (0.5 ml) of protein free urine

3. Boil for 3 to 5 minutes

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 RESULTS
Blue solution – traces (0.5 g/dl)
Cloudy green 1+ ( 1 g/dl)
Yellow orange 2+ ( 1 to 1.5 g/dl )
Orange red 3+ (1.5 to 2 g/dl)
Brick red precipitate 4+ ( > 2 g/dl )

• Other tests –
– Fehling’s test
– Glucose strips

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 GLYCOSURIA ( SUGAR IN URINE )
• Reducing sugars – glucose, lactose, fructose, galactose
• Normal levels 2 to 20 mg / dl in fasting urine
• Glycosuria – clinically detectable amounts
Causes
 Diabetes mellitus
 Hyper thyroidism / pituitarism
 Pregnancy
 Stress / anxiety
 Renal / alimentary glycosuria

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 KETONE BODIES

Acetone , acetoacetic acid & β hydroxy butyrate

Rothera’s test (Acetone , diacetic acid )
Principle : ketone bodies in an alkaline medium react with
sodium nitroprusside to form a calomel ring
Procedure :
 Saturate 5 ml of urine with ammonium sulphate
 Add a crystal of sodium nitroprusside
 Layer liquor ammonia through sides of the test tube
 Permanganate calomel ring at the junction of two layers –
positive for ketone bodies

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 KETONURIA
 Diabetes mellitus
 Fasting , starvation
 Prolonged vomiting

 other tests – Gerhard’s test

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 BILE SALTS
Hay’s test
Principle : bile salts lower the surface tension of urine.
Procedure :
Sprinkle sulphur powder on urine
Sink – presence of bile salts

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 BILE PIGMENTS

Fouchet’s test (Bilirubin )

Principle : BaCl2 combines with sulphates & bile pigments

adhere to these molecules and are oxidised to biliverdin

Procedure :
 Mix 10 ml of urine + 2.5 ml of 10 % BaCl2

 Filter the solution

 Add I drop of Fouchet’s reagent to the filter paper

 Green / blue color – presence of bilirubin

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 BLOOD IN URINE
RBCs in urine – hematuria (bright red )
Blood pigments without RBCs - Hemoglobinuria (brown )
Benzidine test
Principle: ‘Haem’ in hemoglobin acts as a catalyst when hydrogen peroxide is mixed
with Benzidine to form a blue color
Procedure:
♣ Take 2 ml of urine in a test tube
♣ Add 2 ml of glacial acetic acid and knife point Benzidine
♣ Add 1 ml of Hydrogen peroxide to it.
Result : Appearance of blue color – indicates the presence of Hb/ myoglobin

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 HEMATURIA

Causes
♠ Renal diseases – infections, CGN, TB kidney, Nephritic syndrome
♠ Bleeding disorders – leukemia, thrombocytopenia, scurvy, hemophilia
♠ Anti coagulant drugs

HEMOGLOBINURIA
Causes
o Hemolytic anemia
o Snake venom, spider bites, bacterial toxins
o Crush injury, malaria
Other tests – Orthotoludine test, Guiacum test

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 URINE MICROSCOPY

• Procedure

• Mix urine,take 5 ml in a centgrifuging tube

• Centrifuge at 2500 rpm x 15 mins

• Discard the supernatant ,resuspend the deposit

• Take a drop on slide ,cover with cover slip

• Observe under Low power, count under high power

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 Microscopic examination
 Cells – RBCs, Pus cells, epithelial cells
 Casts – hyaline, Red cell, Leukocyte, waxy, fatty, epithelial,
granular casts
 Crystals – acidic urine – oxalate, calcium, uric acid, cystine
alkaline urine – phosphates, urates

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 CAST FORMATION
Formed in lumen of distal and collecting tubules of kidney (acidic
conditions & high solute conc.)

Basic matrix of the cast is Tom Horsfall protein secreted by tubules

Form as a result of
1. Ppt. of gelatin of Tom Horsfall mucoprotein
2. Clumping of cells within protein matrix
3. Adherence of cells or cellular material to the matrix
4. Coagulation of material within the lumen

- Casts dissolve in alkaline urine

- Nearly parallel sides and rounded or blunted ends

- Convoluted, straight or curved

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• Hyaline casts – emotional stress, strenous exercise
• Granular casts – pyelonephritis, lead poisoning
• Epithelial casts – nephrosis ,eclampsia, amylodosis
• Red cell cast – glomerulonephritis,renal infarction
• Fatty cast – degenerating cells
• Waxy cast – chronic renal disease

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Pus cells

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 Urine crystals
• Crystals form by precipitation of urinary
salts.The changes include Ph temperature and
Concentration.
• Based on the Ph of urine crystals can be
classified into acidic ,alkaline crystals

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ACIDIC CRYSTALS ALKALINE CRYSTALS

URIC ACID AMMONIUM BIURATE

LEUCINE,TYROSINE AMMONIUM URATES

CYSTINE TRIPLE PHOSPHATE

Soluble in NaOH, insoluble in HCl Soluble in HCl, insoluble in NaOh

Calcium oxalate and hippuric acid crystals occurs irrespective of urine Ph

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Uric acid

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Hippuric acid

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TRIPLE PHOSPHATE

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AMMONIUM BIURATE

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BILIRUBIN CRYSTALS

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TYROSINE CRYSTALS

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LEUCINE CRYSTALS
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