URINE
EXAMINATION
INDICATIONS OF URINE EXAMINATION
❖ Routine investigation
❖ Diagnosis
Urinary tract diseases
Pregnancy
Endocrine disorders- DM, Hormonal problems
Genetic abnormalities- cystinuria,phenylketonuria
Drug overdose
Carrier of enteric fever
Neoplastic diseases-multiple myeloma
❖ Physical fitness
COLLECTION OF URINE
Container – Wide mouth container
Method of collection –
❖ Single random specimen (routine examination )
❖ 24 hour urine collection (quantitative studies)
Method
Preservative
✔ Formaldehyde 2-4 drops /30ml
✔ Boric acid 0.5gm/60 ml
✔ Thymol 0.1 gm/100ml
✔ Toulene – forms a thin layer on surface,
❖Bacteriological culture –
-Clean catch midstream sample
-Catheterized sample
-Suprapubic aspiration (Infants)
PHYSICAL
EXAMINATION
VOLUME
Normal = 1000 - 2000 ml/24 hr
Polyuria - > 2000 ml/24 hr
causes- Polydipsia, cold, iv fluids administration,
DM, DI, chronic renal failure.
Oliguria - <500 ml/24hr
PRE RENAL - Vomiting, Diarrhea, Excessive
sweating.
RENAL - AGN, ATN, vascular disorders
POST-RENAL- Obstruction
Anuria – < 150 ml/24 hr
causes - Renal ischemia, Haemolytic transfusion reacn
APPEARANCE
COLOR - Pale yellow/ Amber color
❑ Red - Blood, Hemoglobin, Myoglobin, Porphyrin, Beets
❑ Orange - Porphyria,
Rifampicin
❑ Dark Yellow - Conc. urine (fever,inadequate water intake)
Bile pigments and bile salts
Milky White - Chyluria
❑ Black,gray,brown
- Alkaptonuria, Post parental iron administration
& phenol poisoning,
❑ Colorless - Chronic kidney disease
Diabetes insipidus
Diabetes Mellitus
High dilution
CLARITY - Freshly voided urine is clear.
Cloudy - phosphates, urates, fecal contamination by fistula
Turbid –pus cells, RBCs, bacteria, spermatozoa, chyluria
ODOR
Normally urine has faint aromatic odor.
Pungent – d/t Ammonia produced by bacteria
Fruity odor - KA
Mousy odor - Phenylketonuria
Putrid / foul smelling - UTI
SPECIFIC GRAVITY
Definition – Ratio of wt. of fixed volume of solution to
the wt.of same volume of water at a specified
temperature.
Normal S.G.= 1.003 - 1.030
Increase S.G - Decreased fluid intake, fever, vomiting, diarrhea
DM, proteinuria.
Decrease S.G - Diuretics, hypothermia, glomerulonephritis,
Diabetes insipidus
Fixed S.G - CRF 1.010
METHODS
Urinometer
Refractometer
falling drop method
reagent strip
Procedure –
- Fill the glass cylinder ¾ th of
capacity.
-Insert the urinometer with
spinning motion and allow it to
settle.
- Take the reading at lower
meniscus.
CORRECTION FOR
TEMPERATURE
-Urinometer is usually
calibrated at 20o temp.
-Every 30 rise in room temp.
add 0.001 and 30 fall in room
temp. subtract 0.001 to the
above value.
pH- Normal Range – 4.6-8.0 (average 6 i.e slightly acidic)
Causes of acidic urine
Metabolic acidosis
Respiratory acidosis
Diabetic ketoacidosis
Use of acidifiers in T/t of alkaline urine stones
Hypokalemic alkalosis
Causes of alkaline urine
• Respiratory alkalosis
• Metabolic alkalosis
• Alkalinization of urine in T/t of acidic urine stones
• T/t of UTI
• T/t of salicylate poisoning
• Proximal renal tubular acidosis
METHODS - Litmus paper
pH electrode
Reagent strip
Titrable acidity of urine
CHEMICAL
EXAMINATION
PROTEINS IN URINE
Normally < 150 mg /24 hr
Abnormal proteins in urine- Albumin
Globulins (B-J proteins)
Hemoglobins
Myoglobins
CAUSES OF PROTEINURIA
❖Heavy proteinuria – > 3 gm /24hrs
Nephrotic syndrome,
DM,
SLE,
Renal vein thrombosis
❖ Moderate Proteinuria 1-3 gm/24 hr
Chronic GN
Nephrosclerosis
Multiple myeloma
Pyelonephritis.
❖ Mild Proteinuria - < 1.0 gm / 24hr
Hypertension
Polycystic Kidney
UTI
Fever
❖ Microalbuminuria- Excretion of protein 30 mg to
300 mg /24 hrs.
❖ Indicative of Early & possibly reversible renal
damage.
TESTS FOR PROTEIN
Heat coagulation test
Sulpho salicylic acid test
Heller’s nitric acid test
Reagent strip
Heat coagulation test
Principle: Proteins get denatured and coagulated on heating to give
white cloudy precipitates.
Method :
1. Fill 2/3 of the test tube with urine heat only the upper part, keeping
lower part as control.
2. Presence of phosphates, carbonate & proteins give a white cloudy
precipitates.
3. Add 1-2 drops of acetic acid to above white cloudy PPT if
cloudiness persist it indicate presence of protein .
4. If cloudiness disappears it indicate presence of phosphates.
❖ Bence Jones Proteins (B-J
Proteins)
B-J proteins are gamma globulins which are seen
in multiple myeloma & other paraproteinemias.
Method –
Urine is heated under thermal control .
When precipitate appears at 50-60 0C, disappear
on further heating above 90 0C & reappears on
cooling indicates presence of B-J proteins.
This test is now replaced by protein
electrophoresis of urine.
Sulphosalicylic Acid Test
Principle : Addition of sulphosalicylic acid to urine causes formation
of a white precipitate if protein are present (due to acid proteins get
denatured and precipitate is formed).
False positive : gross hematuria ,highly concentrated urine ,excess
uric acid ,sulphonamides and penicillin.
False negative : very dilute urine
Reagent strip test
Principle :
The reagent area of the strip is coated with bromophenol blue
indicator and buffered to an acidic pH which change colour in
presence of protein .
GLUCOSE IN URINE (GLUCOSURIA)–
✔DIABETIC (Glucosuria with hyperglycemia)
✔NON DIABETIC (Glucosuria without hyperglycemia)
• Alimentary
• Renal glucosuria
•Endocrinal & Liver diseases
METHODS –
Reagent strip
Copper reduction test (Benedict’s test)
Benedict’s Test - (Semiquantitative test)
Principle :
Copper sulphate is reduced to cuprous oxide, if Reducing
Substance is present.
Test is not specific for Glucose, other reducing substances
like galactose, lactose, fructose maltose can give positive
results.
Procedure:
- Take 5 ml of benedict’s reagent in a
test tube, add 8 drops of urine and
mix it well.
- Boil over flame for 2 min. allow to
cool .
- Note the change in colour of blue
Benedict's reagent
Reagent strip test
Principle :
Glucose oxidase –peroxidase reaction
• Glucose + oxygen ----->gluconic acid +hydrogen peroxide in
presence of glucose oxidase, hydrogen peroxide with
chromogen gets oxidised and results in color change.
Specific test glucose.
KETONE BODIES
• Ketone bodies – Acetone, Acetoacetic acid and beta hydroxy
butyric acid.
• Causes –DKA, starvation, glycogen storage disease, eclampsia,
severe dehydration, febrile states, after GA
TESTS –
▪ Rothera’s test
▪ Gerhardt’s test
▪ Reagent strip
Rothera’s test:
Principle :
Acetoacetic acid or acetone react with nitroprusside in
alkaline solution to form purple coloured complex .
• Method :
-Take 5ml urine in test tube and saturate with ammonium
sulphate .
- Add a crystal of sodium nitroprusside and mix well .
- Slowly run along the side of test tube ,liquor ammonia to
form a layer ring.
❖ Gerhardt’s Test-
10% ferric chloride react with ketone
bodies in urine & urine become reddish
or purplish if Acetoacetic acid is
present.
Certain drugs like salicylate & L-dopa may
give similar results so is not specific.
❖ Reagent Strip test-
Is modification of Rothera’s test-
Bile Derivatives in urine
► Urobilinogen
► Bile Pigments – Bilirubin & Biliverdin.
► Bile Salts – Cholic acid, Chenodeoxy cholic acid
& Lithocholic acid.
► Urobilinogen - Normally excreted in
urine in small amount.
► Increased urobilinogen seen in Hemolytic
jaundice and in hepatocellular jaundice.
Ehrlich Aldehyde Test
►Principle – Urobilinogen combines with Ehrlich
reagent to give purple coloured compound.
►Procedure –
10 ml urine + 1 ml Ehrlich reagent
Wait for 3-5 min
Purple coloured complex
Interpretation - Development of red purple
colour indicates presence of urobilinogen.
BILE SALTS AND BILE PIGMENTS
❖ Bile Pigments :
✔ Gmelin’s Test for bile pigment
✔ Harrison Fouchet test for bile
pigment
✔ Foam Test
Fouchet’s test
It will detect the bile pigments in the urine.
Reagents are:
Barium chloride 10%
Fouchet’s reagents consist of:
Trichloracetic acid = 25 grams.
Ferric chloride (FeCl3) 10% = 10 mL
Distiled water = 100 mL
Procedure of Fouchet’s test:
5 ml urine + 1 to 2 drops of glacial acetic acid
(concentrated) + 2.5 ml of barium chloride (BaCl2 10%).
Mix well, the sample will become cloudy.
Filter or centrifuge to obtain the precipitate.
To this precipitate add one drop of Fouchet’s reagent
(Ferric chloride).
Principle :
When ferric chloride is added to urine in trichloroacetic
acid, it oxidizes bilirubin to green color biliverdin.
Immediately blue-green color develops indicating
positivity for bilirubin.
Hay’s Sulphur test for bile salts:
- Take 2 ml urine sample in a container & carefully sprinkle the
sulphur powder over the surface of urine.
(Also run water as the control).
Principle –
Bile salts reduce the surface tension so sulphur granules
sink to the bottom.
Interpretation -
In the case of presence of bile salts, sulphur powder will
settle in the bottom.
In the case of absence of bile salts, sulphur granules will
remain over the surface.
BLOOD IN URINE
HEMATURIA – Acute nephritis, PN, Urinary calculi, renal TB,
embolic nephritis, leukemias, blood dyscrasias, benign and malignant
tumor of kidney, drugs.
HEMOGLOBINURIA – Intravascular hemolysis, severe burns,
hemolytic transfusion reaction, black water fever, AIHA, G6PD
deficiency, PNH, drugs.
Benzidine test
Method-
• Make saturated solution of benzidine in glacial acetic acid
. • Mix 1 ml of this solution with 1 ml of hydrogen peroxide
in a test tube .
• Add 2ml of urine to above test tube .
Interpretation-
• If green or blue colour developed within 5 min the test is
positive .
False positive: contamination of urine by oxidising agent.
False Negative :presence of reducing agent like ascorbic acid
or formalin.
Other tets for hematuria
Reagent strip test : uses chromogen –o- toluidine
Microscopy method use to detect RBCs in urine
Microscopy
► Cells - WBCs, RBCs, Epithelial
cells
► Casts – Hyaline cast, Granular
casts, Cellular casts
► Crystals
URINE REPORT
Name: Regd no:
Age: Sex: Time of collection:
Chief complaints:
PHYSICAL EXAMINATION
Appearance-Colour,clarity :
Odor :
Volume :
Specific gravity :
CHEMICAL EXAMINATION
Reaction :
Proteins :
Sugar :
Ketone bodies :
Bile salts and pigments :
Urobilinogen :
MICROSCOPY
RBCS :
PUS CELLS :
EPITHELIAL CELL :
CASTS :
CRYSTALS :
OTHER FINDINGS :
IMPRESSION
Case 1-
► 8 year male child presented
with pedal edema, puffy face &
easy fatiguability.
► Urine examination report---
► PHYSICAL EXAMINATION
► Appearance : Turbid
► Odor : Aromatic
► Colour : Whitish
► Specific gravity : 1.040
► CHEMICAL EXAMINATION
► Reaction : Acidic
HEAT COAGULATION TEST : POSITIVE
Interpretation- ?????
► PHYSICAL EXAMINATION
► Appearance : Turbid
► Odor : Aromatic
► Colour : Whitish
► Specific gravity : 1.040
► CHEMICAL EXAMINATION
► Reaction : Acidic
HEAT COAGULATION TEST : POSITIVE
Interpretation - PROTEINURIA
Case 2
► 45 year obese female brought
to causality with altered
sensorium.
► BSL (R) – 440 mg/dl
► Urine examination done…
► PHYSICAL EXAMINATION
► Appearance : Clear
► Odor : Fruity
► Specific gravity : 1.045
► CHEMICAL EXAMINATION
► Reaction : Acidic
► BENEDICT’S TEST : POSITIVE (Orange ppt)
► Interpretation ??????
► PHYSICAL EXAMINATION
► Appearance : Clear
► Odor : Fruity
► Specific gravity : 1.045
► CHEMICAL EXAMINATION
► Reaction : Acidic
► BENEDICT’S TEST : POSITIVE (Orange ppt)
► Reagent strip test : Shows brown color (3+)
► ROTHERA’S TEST : POSITIVE
► INTERPRETATION ?????
► PHYSICAL EXAMINATION
► Appearance : Clear
► Odor : Fruity
► Specific gravity : 1.045
► CHEMICAL EXAMINATION
► Reaction : Acidic
► BENEDICT’S TEST : POSITIVE
► Reagent strip test : Shows brown color (3+)
► ROTHERA’S TEST : POSITIVE
► INTERPRETATION :
GLYCOSURIA WITH KETONURIA.
► 50 year male presented with weight loss, nausea,
vomiting and yellowish discoloration of sclera.
► Total Bilirubin – 4.5 mg/dl
► Urine examianation done
► PHYSICAL EXAMINATION
► Appearance : Turbid
► Color : Dark yellow
► Specific gravity : 1.040
► CHEMICAL EXAMINATION
► Reaction : Acidic
FOUCHET TEST : GREEN PPT
HAY’S SULPHER TEST : SULPHER POWDER
SINKS TO BOTTOM.
Interpretation- ?????
► CHEMICAL EXAMINATION
► Reaction : Acidic
FOUCHET TEST : GREEN PPT
HAY’S SULPHER TEST : SULPHER POWDER
SINKS TO BOTTOM.
Interpretation- BILE PIGMENTS AND
BILE SALTS
ARE PRESENT.
