URINE EXAMINATION

1.WHAT ARE METHODs OF URINE COLLECTION

Early morning sample-qualitative,.more acidic-preserve the cast ,cel

Random sample- routine purpose
Midstream clean catch sample-UTI
Post prandial sample-D.M
Catheterised-ininfants,bedridden patients
Suprapubic needle aspiration
24- hour specimen: Quantitative estimation of proteins, hormones and electrolytes

2.WHAT ARE PRESERVATIVES USED FOR URINE COLLECTION

Sample should be examined within 1-2 hrs ofvoiding.

If delay is expected, sample can be kept in refrigerationfor max 8 hrs.
Not recommended for routine analysis as they interfere with reagent strip techniques and
chemical testfor protein.
Preservatives for 24-hour urine sample:
1. Hydrochloric acid: Used when detecting adrenaline, noradrenaline, vanillylmandelic
acid (VMA) and steroids.
2. Toluene: It forms a thin layer and hence physical barrier against bacteria and air.
3. Boric acid: General preservative (sample can be kept for 24 hours without
refrigeration)
4. Thymol: Inhibits bacteria andfungt.
5. Formalin: Excellent for preservation offormed elements. 6-3 daop ot *o i

3.NORMAL URINE ANALYSIS VALUES

.

Color- Yellow (light/pale to dark/deep amber) Urobilirubin -Small amount (0.5

Clarity/turbidity-Clear or coudy 1 mg/dL)
pH-4.5-8 Blood-s3 RBCs
Specific gravity 1.005-1.025 Protein-s150 mg/d
Glucose-s130 mg/d RBCs-s2 RBCs/hpf hpf
Ketones- None WBCs-S2-5 WBCs/hpf
NitritesNegative Squamous epithelial cells - s15-

Leukocyte esterase-Negative 20 squamous epithelial cells/hpf

Crystals-Occasionally *
Casts-0-5 hyaline casts/pf
Bilirubin- Negative +Bacteria -None

4.PHYSICAL EXAMINATION FINDINGS

Volume: Color; appearance; Odour; Ph; Specific gravity

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 5.VOLUME ABNORMALITIES

Normal-1-2.5 L/day
Oliguria- Urine Output <400ml/day
Seen in
Physiological-Dehydration, hot climate
.Pathological-Shock
Acute glomerulonephritis
-RenalFailure
Polyuria-Urine
Seen in
Output> 2.5 LIday
Physiological-Increased water ingestion, pregnancy, cold climate
Pathological-Diabetes mellitus and insipidus.
Chronic renal failure (Loss of concentrating ability of renal tubules)
Diuretic therapy
*
Anuria-Urine output< 100m/day
Seen in
renal shut down
Acute tubularnecrosis
Complete urinary tract obstruction
Nocturia-excretion of urine by a adult of >500ml
with a specific gravity of <1.018
6.cOLOUR

Normal- pale yellow in color due to pigments urochrome, urobilin and

Abnormal uroerythrin.
Colourless- dilution, diabetes mellitus, diabetes insipidus, diuretics
Milky- purulent genitourinary tract infection,
Yellow to orange-jaundice,rifampicin(orangish red)chyluria,lipiduria Pifopun
Red-beetroot ingestion,haematuria, hemoglobinuria beetrd
Brown/ black- alkaptunuria, melanin
blue to green- Pseudomonas infection Psudomona

7.APPEARENCE
Normal-clear
Abnormal boctena - fiCams

Turbidity-precipitation of crystals or nonpathologic Salts or by leukocytes, bacteria, chyluria

(filariasis), lipiduria(nephritic syndrome)
Smoky urine RBCs
Hazy urine- Mucous
Sme
Pseudochyluria- use of paraffin based vaginal creams in treatment of Candida infection
Todifferentiate chyluria from lipiduria-ether
extraction test. if extacted its chyluria. not extracted
ipiduria

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8.ODOUR

Nomal: Aromatic due to the

Abnormal:
volatile organic acids
On long standing ammonical (decomposition of urea forming ammonia which gives a
-

strong ammonical smell)

Foul, offensive - pus or inflammation
Sweet Diabetes
Fruity Ketonuria
Maple syrup-Hike - Maple Syrup Urine Disease

Rancid-Tyrosinaemia
Fishy-UTl by Proteus,
Cabbage- Methionine malabsorption
Sweatyfeet-Isovalericacidemiaand glutaric acidemia
Rotten eggs-Cystinuria
.***

9. REACTION-PH

Urine pH ranges from 4.5 to 8 neural- pupe

Tested by:
1.Reagentstrip method?
Indicators methyl red and bromothymol blue give a range of orange, green,
and blue colours as the pH rises, permitting estimation of pH values to within half
a unit within the range of 5 to 9.
2. Litmus paper test
Litmus paper test: Blue litmus paper turns red, in acidic urine and Red litmuss
turns blue in alkaline urine
3.pHmeter
Electrode of pH meter is dipped in urine and the reading is taken

Causes of Acidic urine

Ketosis-diabetes, starvation,fever
Systemic acidosis
UTI by E.coli 9
Acidification therapy
High protein diet
+Causes of Alkaline urinee
Strict vegetarian
Systemic alkalosis
UTI by pseudomonas or Proteus
Alkalization therapy
10.SPECIFIC GRAVITY

It is measurement of urine density which reflects the abilityofthe kidney to concentrate or dilute
the urine
Simplytelativeamount of solutes present in the urine
Normal value: 1.015-1.025..
Measured by:

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 1. Urinometer
Assessing specific gravity of urine at room temperature which is
principle of BUOYANCY based on the
requires minimum of 15 ml of urine
Correction of 0.001 should be made for each
3° Crise or fall in temperature.
2.Refractometer-drops of urine adequate; accurate method
3.Dipsticks
Increased specific gravity-Hypersthenuria uewal Menisus
0.002
Diabetes melitus
Dehydration Remeve frutty
REäiag léesve

Proteinuria (Nephrotic syndrome)

Glycosuria bu RHer popes Speciio gray
Lowwater intake
Decreased specific gravity- Hyposthenuria <1.007 Une in qylinder

Polydipsia
piabetes insipidus
Diuretics drometer
Early stages of Chronic Kidney Disease

Isosthenuria: - Mercury bulb

Fixed specific gravity at 1.010 (occurs in Chronic Kidney

Disease)

11. TEST FOR PROTIEN

Normal protein excretion-Up to 150

mg/24 hours
Tamm-Horsfall protein-(secreted by the distal tubular cells and ascending loop o Henle dsaP
constitutes 1/3rd or more of the total urinary protein loss.
Co,PO4
TEST-HEAT COAGULATION TEST
Coaguloo

Principle
proteins are denatured & coagulated on heating to give Cahoa
white cloud precipitate.
Method:
take 2/3 of test tube with urine, heat min-nitnc
only the upper part keeping lower part as control.
Presence of phosphates, aied
carbonates, proteins gives a white cloud formation. Add 1-2
10% acetic acid, if the cloud persists it indicates it is protein drops of
carbonates /phosphates. (acetic acid dissolves the

Interpretation/grading: Acid metapro tln

Negative - No turbidity or cloudiness.
Trace-Cloudiness visible against a black background (5 mg/ dl). Jutefese wit
1 6 o c g l v m
 1t-Definite cloudiness without flocculation and granularity (10-30 mg /I dl ). f¬tultn,
2+-Heavy and granular cloudiness without flocculation. (40-100 mg/ dl).
3t- Dense
opaque cloud with marked
4+ Thick cloudiness with precipitationflocculation ( 200-500 mg dl).
/

Other Tests for Protein

Nitric acid test
Sulphosalicylic acid test
Test with Esbach's reagent
Protien reagent strip- Variable shades of green develop if protein is present in urine.
Quantitative method botnephond
Esbachs metheod (( Picnc aca d + Cuhic aid
Auerbach method

CAUSES OF PROTEINURIA
MINIMAL PROTEINURIA(<1 gm/day)
MODERATE PROTEINURIA (1-4 gm/day)
Exercise
Feve Nephrosclerosis
Acute glomerulonephritis
Emotional stress Toxic nephropathies nepwrosdecum
Chronic Pyelonephritis Pre-eclampsia
NephrosclerosisS
Chronic interstitial nephritis
Postural proteinurias
Transient proteinurias
SEVERE PROTEINURIA (> 4 gm/day)

Nephrotic syndrome
Diabetic nephropathy, severe
Renal amyloidosis
Lupus nephritis
Toxemia of pregnancy
Selective proteinuria
Non-selective proteinuria
When LMW proteins like albumin (MW-66000) or
transferrin(MW-76000) are selectively excreted When HMW protein like globulin,
through kidney. fibrinogen in addition to LMW
protein are excreted through
Eg- all causes of nephrotic syndrome.
kidney
Microalbuminuria
presence of albumin in urine above normal level bt below detectable range of conventional urine
dipstick method
Defined as urinary excretion of 30 to 300 mg/24 hrs of albumin in urine.
an important prognostic marker for kidney disease
indiabetes mellitus (earliest sign of renal damage in DM)
in hypertension
Bence Jones proteins of iabehi eplkropatuy
Cag ig
Candio Vascuular mootalty
Remave Cequtm
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Conh n e boi&g

TP
 These are
monoclonal immunoglobulin light chains (kappa or lamda) synthesized by
plasmacells neoplastic
Test-Themal method
Proteins precipitating at
40-60c &then dissolving when the urine is brought to boiling(100c) &
reappearSwhen the urine is cooled.

12.TEST FOR GLUCoSE

malquaur Oyhonma.
Pbhapwteinenmo
Test-BENEDICT'S TEST(semiquantitative) Obsunte BJP does nt nule outmuship e
Principle- benedict's reagent contains cuso4.In the presence of
converted to cuprous oxide which is hastened reducing sugars cupric ions are
by heating, give the color.
to
Components of Benedict's reagent
Sodium carbonate- 100 gm (Provides alkaline conditions which are required for the redox
reaction)
Sodium citrate- 173 gm (complexes with the copper (1) ions so that they do not deteriorate to
copper() ions during storage)
Copper sulphate- 17.3 gm
Proten wilinferera wit CSo4 ? zo do
Method-: heat Caq n
Take 5ml of Benedict's beore
reagent .Boil for 3-5 minutes(look for auto
drops)of urine. .Boil for 2 minutes.Cool and note the colour mixture. reduction). Add 0.5ml (8
Interpretation: positive test indicate presence of reducing sugar
Blue= negative
Yellow=+(<0.5%)
green=++(0.5-1%)
Yellow-orange=+++(1-2%)
Brickred=++++(>2

Sugars detected by Benedict's test

Glucose False+
Galactose Ascorbic acid,
Lactose Salicylates
Fructose
Maltose Gucoreic. ccid ***

Homoguitis ocid
Pentose
To
dopo
confirm it is glucose,
Reagent Strip method-dip stick dipsticks can De usea (giucose oxiaasej
Principle:
Based on a specific glucose oxidase and
Chemistry: peroxidase method
Glucose+ 02 glucose oxidase Gluconic acid + H202
H202+Chromogenperoxidase Oxidized chromogen + H20

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Other Methods of detecting glycosuria
Fehling'smethod
Osazone test

Causes of Glycosuria

Glycosuria usually occurs when the blood level is greater than 180 -
200 mg/d
o
Renal Glycosuria (due to low renal threshold- Acquired/Congenital) e.g.
Fanconi's Syndrome
o
Gestational Glycosuria (Reduced Renal threshold)
o
Alimentary glycosuria -

On consuming high amounts of carbohydrates

13.Ketone bodies
Conhicos koid, Pifuitot
Defect in carbohydrate metabolism or
absorption or an inadequate amount of carbohydrate in
the diet-the body compensates
by metabolizing increasing amounts of fatty acids.ketone
bodies are are products of fat metabolism
I n ketonuria, the three ketone bodies
present in the urine are acetoacetic (diacetic) acid (20%),
acetone (2%), and 3-hydroxybutyrate (about 78%).
Causes of ketonuria
DM
STARVATION
DEHYDRATION
Methods:
Rothera Method;
Principle-acetone &acetoacetic acid react with sodium nitroprusside in the presence of
alkali to produce purple colour.
Method:Saturate about 5ml of urine with amm nium sulphate. Add few crystals of
sodium nitroprusside and shake well. Add liquor ammonia
through the sides of the test
tube.Formation of a purple ring at the junction indicate a positive test.
Other test
1. Reagent Strip method
2. Nitroprusside Tablet test
3 Gerhardt ferric chloride test (Only for acetoacetic acid)
14.TEST FOR Bilirubin
I t is breakdown product of Hemoglobin.
unconjugated Bilirubin& conjugation
Normal adult urine has only,0.02 mg/dl of Bilirubin
Increased urinary Bilirubin (only
CONJUGATED
BILIRUBIN Joccurs in-
o Obstruction to bile outflow from liver
o
Hepatocellular disease with decreased excretion of conjugated Bilirubin in bile

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 Bile pigments
Normal urine- Abnormal urine
O Bilirubin
OUrochrome
OTraces of Urobilin OUrobilinogen
O Biliverdin
OUrobilin
.Urobilinogen
Conjugafed bilirubin reaches eventually to duodenum from liver but without
absorbed, passes on to colon where it is acted getting
Normal output of urobilinogen in urine is upon by bacteria to form Urobilinogen
Test- ehrlich test 0.5 to 2.5 mg/24 hours
In 5 ml of urine
add 0.5 ml of Ehrlich's reagent(HCI 20 ml, d/w 80 ml,
amino benzaldehyde 2 gm). Allow to stand for 5 min. development of p-dimethyl
pink colour
indicates +test.
Causes-hemolytic jaundice, early hepatitis, hepatocellular jaundice.
Bile salts
Primary bile acids
Secondary bile acids
Cholic acid and chenodeoxycholic Deoxycholate and lithocholate, are
acid (CDCA)- formed in the colon
conjugated with glycine or taurine, Sodium taurocholate and sodium
glycocholate

Hay'ssulphur test
In 5 ml of urine sprinkle a
pinch of sulphur particles. If bile salt is present sulphur
particles will sink to the bottom because bile salts lowers the surface
Control test-take distilled water tension of urine.
15.Blood in urine

*1.Test- BENZIDINE TEST

o
Principle-The peroxidase activity of hemoglobin decomposes
hydrogen peroxide
releasing nascent oxygen which in turn oxidizes benzidine to give blue color.
o Method-mix 2ml of benzidine solution with
2ml of hydrogen peroxide in a test tube.
Take 2ml of
urine & add 2ml of above mixture. A blue or green color within 5 min indicates +
reaction
2.Blood/ Heme proteins Reagent Strip method
o Principle:
o liberation.of oxygen from peroxide in
haem in free haemoglobin of
reagent strip by peroxidase like activity of
lysed RBC or myoglobin
o
If green spot is present then intact RBCs are present. If uniforim
green colour is
present then it indicates the presence of haemoglobin
or myoglobin
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