ANALYSIS OF ABNORMAL

CONSTITUENTS OF URINE
INTRODUCTION : -

 The normal glomerulus filters plasma and retains most of

the large molecular weight and useful substances.
 Further, the small molecules filtered are also reabsorbed by
renal tubules.
INTRODUCTION : -

 In many pathological conditions, the urine may contain

abnormal constituents which are not excreted normally-
 due to defective filtration by glomerulus
 or impaired reabsorption by renal tubules.

 Presence and analysis of these abnormal constituents in

urine will help in the diagnosis of pathological condition.
URINE EXAMINATION

 PHYSICAL EXAMINATION

 CHEMICAL EXAINATION

 MICROSCOPIC EXAINATION
PHYSICAL EXAMINATION

1. VOLUME
2. COLOUR
3. APPEARANCE
4. ODOUR
5. pH
6. SPECIFIC GRAVITY
 PHYSICAL EXAMINATION
 VOLUME:
 Normal urine output is 800-2000 ml/day
 POLYURIA: Urine Output > 2.5 L / day; seen in -
 Diabetes Mellitus , Diabetes Insipidus
 Diuretics , later stages of renal failure
 OLIGURIA: Urine Output < 400 ml/day ; seen in -
 Acute Nephritis
 Excess fluid loss due to vomiting , diarrhea ,fever, haemorrhage , cardiac failure
 ANURIA: Complete cessation of urine. Urine Output < 50ml/day ; seen
in -
 Shock , acute tubular necrosis , Bilateral renal stones , mismatched blood transfusion
 Ratio of urine output in day and night is 2:1 to 3:1
 The ratio is reversed or decreased in renal diseases
 High urine output at night (NOCTURIA) is one of the early
symptoms of chronic renal disease.

Appearance :
 Normal urine is clear and does not contain any
sediments.

 Turbid urine is seen in

 1. urinary tract infections (excretion of pus cells) and
 2. protein in urine.
 3. On long standing ,urine turns turbid due to
precipitation of phosphates present in urine.
 COLOUR :
 Normal colour is pale yellow / amber yellow due to excretion of urochrome or
urobilinogen.
 Dark coloured urine is -due to increased concentration of urine , as in acute
glomerulo nephritis.
 Red colour- seen in Haematuria , due to stones in urinary tract, carcinoma of
bladder, injury to urinary passage , acute glomerulo nephritis)

Reddish brown – Haemoglobinuria ( incompatible blood transfusion)

Myoglobinuria ( road traffic accidents, crush injury)

 Yellow colour- seen in Jaundice due to excretion of bile pigments.

 Milky white urine or Chyluria is seen in –– Filariasis

 Black to brown – seen in Porphyrias, PKU, Alkaptonuria.
 ODOUR :

 Normal odour is Faint Aromatic odour ( due to volatile

organic acids )
 Sweety / fruity odour – seen in Diabetic keto acidosis
 Mousy odour - seen in Phenyl ketonuria
 Cabbage odour - seen in Tyrosinemia
 putrid or ammonical odour / Foul smell – is due to bacterial
decomposition, if urine is kept outside for a long time.
 Burnt sugar odour- seen in Maple syrup urine disease
 Specific gravity
 Indicates concentrating ability of kidneys
 Ranges from 1.015 – 1.025 for normal urine
 Measured by Urinometer
 S.G α Solutes
Volume
 Increased - in Diabetes mellitus (glucose in urine ) low water
intake, albuminuria ( albumin in urine) , Chronic renal failure
 Decreased - in diabetes insipidus , high fluid intake
 Isosthenuria - fixed specific gravity (1.010 )– seen in later stages of
renal disease. (chronic and acute kidney failure ) , sickle cell trait .
MEASUREMENT OF SPECIFIC GRAVITY

Urinometer
CORRECTED SPECIFIC GRAVITY
 Specific gravity = observed specific gravity + temperature correction

 Temperature correction = Room temperature – calibrator temperature X 0.001

3
 Ex- Specific gravity = 1.016 +{ 36 – 27 / 3 X 0.001}
 = 1.016 + 0.003
 = 1.019
 pH :

 Freshly voided urine has a pH 4.6-8 (mean pH 6.0 )

 On long standing becomes - alkaline due to bacterial action of
conversion of urea to ammonia and increases the pH
 Urine excreted after meals is alkaline and is known as alkaline
tide.
 Fruits and veggies – alkaline urine
 High protein diet – acidic urine
 In D.K.A – acidic urine ( due to Ketone bodies - acetoacetic acid
& beta hydroxy butyric acid )
MEASUREMENT OF URINARY PH
Urinary pH is measured by-
 pH papers

 Litmus papers
ABNORMAL CONSTITUENTS OF URINE

1. Reducing sugars

2. Ketone bodies

3. Proteins

4. Blood

5. Bile salt

6. Bile pigments.
 TEST FOR REDUCING SUGARS –
BENEDICT’S TEST

EXPERIMENT OBSERVATION INFERENCE

Take 5ml of Brick red color Indicates the
Benedicts reagent precipitate is presence of
in a test tube, add 8 observed. reducing sugars in
drops of urine the given urine
sample & boil for 2 sample.
minutes.
 COMPOSITION

1 liter of Benedicts reagent contain

1)100 g of Na2co3

– provides alkaline medium

2) 173g of sodium citrate

– keep copper in ionic state

3) 17.3 g of cuso4 – provides cu ions

 PRINCIPLE :

In alkaline medium, (sugars behave as weak acids &)

tautomerise to form enediols which are powerful reducing
agents , these enediols reduce the cupric (Cu2+)ions to red
color cuprous (Cu+) oxide.
 Benedicts test is a semi quantitative test:
 The colour of the precipitate

indicates the approximate

concentration of

the reducing sugars.

Blue color Negative test absence of sugar

Green precipitate Positive test 0.5g% (+)
Yellow precipitate Positive test 1.0g% (++)
Orange precipitate Positive test 1.5g% (+++)
Red precipitate Positive test 2.0 g% (++++)
Brick red precipitate Positive test > 2.0 g%(++++)
TESTS FOR REDUCING SUGARS
Clinical Interpretation:
 Glycosuria – is excretion of reducing sugars in urine.
 Glycosuria is a non-specific term.
 Any reducing sugar found in urine is denoted by glycosuria

( mostly glucose)
 Galactosuria - in galactosemia
 Fructosuria - in hereditary fructose intolerance
 Pentosuria - in essential Pentosuria

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 1.Determination of Glucose.

Benedicts test

Fehling's test
 2. Determination of Lactose

Yeast fermentation test

Osazone test.

Rubners test.
 3. Seliwanoffs Test for Fructose
 4. Bial’s Test For Pentoses
 GLYCOSURIA - CAUSES

a. Hyperglycemic glycosuria - Diabetes mellitus ,

Hyperthyroidism, hyperpituitarism and hyperadrenalism

b. Non- Hyperglycemic glycosuria or Renal glycosuria

- Pregnancy, hereditary diseases of renal tubules(Fanconi

syndrome ) , heavy metal poisoning , glomerulosclerosis .

c. Alimentary glycosuria - transient glycosuria

d. Stress, severe infections, increased intracranial pressure

GLYCOSURIA

Examples of non-carbohydrate substances which give a

positive Benedict's reaction are:

a) Creatinine

b) Ascorbic acid

c) Glucuronates

d) Drugs: Salicylates, PAS and Isoniazid

 TEST FOR KETONE BODIES-
ROTHERA’S TEST
EXPERIMENT OBSERVATION INFERENCE
Take 5ml of urine, Permangate or Indicates the
saturate with purple colour ring presence of
Ammonium sulphate is observed at the ketone bodies
crystals. Add 2-3drops junction of two ( Acetone ,
of freshly prepared 5% layers acetoacetate) in
Sodium nitroprusside the given urine
& mix well. Then add sample.
2ml of strong liquour
Ammonia slowly
along the sides of the
test tube
Rothera’s Test

Principle:
Nitroprusside in alkaline medium reacts
with a ketone group to form a purple ring.
 It is given by acetone and acetoacetate,
but not by Beta hydroxy butyric acid.

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Acetone, acetoacetate and beta hydroxy butyrate are the ketone bodies.
They serve as energy source for tissues like brain , muscle during
starvation .
Normal blood level < 1 mg/dl ; Normal urine conc. < 1 mg/day
 Ketonemia and hence ketonuria occurs mostly in conditions of glucose
deprivation.
Ketosis = ketonemia + ketonuria

Causes of Ketonuria:

1) Pathological - Uncontrolled diabetes mellitus , Toxemia of pregnancy

2) Physiological - Starvation , High fat feeding , Heavy exercise

OTHER TESTS FOR K.B
 Rothera's test for acetone.
 Gerhard's test for acetoacetic acid

 Tablet test
TEST FOR BLOOD – O-TOLIDINE TEST
EXPERIMENT OBSERVATION INFERENCE

Take 1ml of Development of Indicates the

O-tolidine in a T.T blue color presence of Blood in
and add 1ml of 1 in 10 indicates presence urine
H2O2 . Mix well. of blood pigment.
Make it into 2 parts The second tube
into another TT. serves as control.
To 1 part add 1ml of
given urine solution.
To the other TT, add 1
ml of D.H2O ,as
control.
PRINCIPLE:

 Hemoglobin of blood decomposes Hydrogen peroxide

in presence of the enzyme catalase – liberating water and
nascent oxygen.
 Nascent Oxygen oxidizes O-Tolidine to a blue or green
colored Tolidine blue. This color changes to brown with
in a few minutes on exposure to air.
Clinical Interpretation:

Hematuria : Presence of blood in urine.

a. Gross hematuria:

b. Microscopic hematuria:
 b. Microscopic hematuria:
a. Gross hematuria: Blood is not visible to naked
Urine appears reddish in eyes.
gross hematuria. It is observed in:
observed in : Malignant hypertension,

Trauma, Sickle cell anemia,

Renal stones, Coagulation disorders,

Malignancies, Polycystic kidney disease,

Tuberculosis

acute glomerulonephritis.

Hemoglobinuria - free Hb. in urine imparts smoky red color –seen in

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Incompatible blood transfusion , malaria, Auto immune hemolytic
anemia.
OTHER TESTS
 Guaiac test
 TEST FOR PROTEIN-
HEAT COAGULATION TEST
EXPERIMENT OBSERVATION INFERENCE

Take urine solution White coagulum Indicates presence

up to 3/4th of TT, is observed at the of protein in the
&heat the upper top of the test given urine sample.
part. tube.
Add 2-3 drops of
10% Acetic acid
and again heat the
upper part of the test
tube.
 Principle:
 Albumin is denatured and coagulated on
heating at the iso-electric point( acetic
acid )
 Acidification is necessary because-
- In alkaline medium, heating may
precipitate carbonates and phosphates.
Acetic acid dissolves this precipitate.
The heated upper half shows
turbidity due to the precipitation
of proteins, the lower half serves
as a control for comparison.
 TEST FOR PROTEIN-
SULPHOSALICYLIC ACID TEST
EXPERIMENT OBSERVATION INFERENCE

Take 3ml of urine White precipitate Indicates presence

solution in a test is observed. of protein in urine
tube,
Add 3 drops of 20
% sulphosalicylic
acid .
 Principle
 In acidic medium, proteins carry positive charges which is
neutralized by negatively charged alkaloidal reagents, like
sulphosalicylic acid and the proteins get precipitated.
 TEST FOR PROTEIN-
HELLERS NITRIC ACID TEST
EXPERIMENT OBSERVATION INFERENCE

Take 3ml of urine A white ring is Indicates presence

solution in a test formed at of protein in urine
tube , junction of two
add1ml of layers.
Conc.HNO3 along
the walls of the test
tube.
  Principle:
 Concentrated HNO3 causes denaturation and hence
precipitation of proteins.
Clinical Interpretation-
Normally protein excretion in urine is in insignificant
amount ( < 20-80 mg/day) .
This small amount is not detectable by routine methods.
Proteinuria : presence of protein in urine.(in detectable
quantities)
The most common type of proteinuria is albuminuria;
hence proteinuria and albuminuria are used synonymously
Proteinuria.

It can be caused by-

a) Increased glomerular permeability

b) Reduced tubular reabsorption

c) Increased secretion of proteins

d) Increased concentration of low molecular weight

proteins in the plasma
Proteinuria may be- Physiological or Pathological

Physiological Proteinuria

Severe exercise , Pregnancy , Postural , After high protein

diet , Exposure to cold

Pathological proteinuria

I. Pre Renal:
 Severe dehydration , Heart disease , Ascites (due to
increased intra-abdominal pressure) , Severe anemia,
and Fever
II. Renal: All inflammatory, degenerative or destructive diseases of
kidney; the most common ones are:
 Nephrotic syndrome, Pyelonephritis , Acute and Chronic
glomerulonephritis , Nephrosclerosis , Tuberculosis of kidney
 Renal failure.

III. Post Renal – due to inflammation of lower urinary tract

 Severe urinary tract infections
 Inflammatory, degenerative or traumatic lesions of pelvis, ureters,
bladder, prostate or urethra
 Bleeding in genito urinary tract
 Pus in urine
 BENCE JONES PROTEINS
 Bence Jones proteins are light chain immunoglobulins
 Excreted in urine of a patient suffering from multiple

myeloma.

 These proteins are detected in urine by classical heat test :

 These proteins precipitate between 40-60 degree centigrade
 Upon further heating, turbidity disappears to reappear on
cooling
 These proteins redissolve on boiling unlike albumin
 TEST FOR BILE SALTS-
HAY’S SULPHUR TEST
EXPERIMENT OBSERVATION INFERENCE
Take Two test tubes Sulphur powder Indicates the
label as Test sinks in TEST presence of Bile
(T) ,Control (C). salts in the given
Take 5ml of urine in Floats in urine sample.
a test and 5ml of CONTROL
Distilled water in
control.
Sprinkle little
Sulphur powder in
both test tubes

test Control
 Principle:
 Bile salts lower the surface tension allowing the sulphur
powder to sink
 Clinical Interpretation:
 Bile salts – Na and K salts of Glyocholic and
Taurocholic acids.
 Excreted in urine –in obstructive jaundice due to
regurgitation of bile into blood . ( causes are - due to gall
stones, cancer of head of pancreas ) –
 Other tests- Peten koffer`s test.
 TEST FOR BILE PIGMENTS-
FOUCHET’S TEST
EXPERIMENT OBSERVATION INFERENCE
Take 5 ml of urine in Green color is Indicates the
a test tube. observed on the presence of
Add 1ml of 10 % filter paper. Bile pigments in
Bacl2 & 1 ml of the given urine
Mgso4 . mix well, sample.
wait for 5 minutes
and filter it.
Dry the precipitate on
the filter paper.
Add to the
precipitate, few drops
of fouchets reagent.
  Principle:
 BaCl2 reacts with Mg.sulphate in urine to form
precipitate of barium sulphate, which adsorbs bile
pigments in urine .

( if present )

- Composition of Fouchets reagent – FeCl3 in TCA

 The adsorbed bile pigments are released by
TriChloroAceticacid (TCA) of Fouchets Reagent and
FeCl3 oxidizes bilirubin to biliverdin (green color).
Clinical Interpretation

Bilirubin in urine means increased amount of conjugated

bilirubin because unconjugated bilirubin is water insoluble and
is also bound to albumin, hence cannot cross the glomerular
membrane.

Causes of bilirubinuria are: hepatic and obstructive jaundice

1) Moderate to severe hepatocellular damage

2) Obstruction of bile duct- Intra or extra hepatic

In prehepatic jaundice, bilirubin is absent in urine.

(unconjugated bilirubin– seen in hemolytic jaundice)
OTHER TESTS
 Foam test
 Gmelin's test , SMITH`S TEST

 Ehrlich's aldehyde test – urobilinogen

 Schlesingers test
ABNORMAL URINE REPORT
 1. PHYSICAL CHARACTERS
 VOLUME
 APPEARNCE
 COLOR
 ODOUR
 pH
 SPECIFIC GRAVITY
2. CHEMICAL CHARACTERS
Procedure Observation Inference

1. Test For Reducing Sugars – Benedicts

2. Test For Ketone bodies – Rotheras

3. Test For Proteins– Heat Coagulation

test +2

4. Test For blood – benzidine / O-toluidine

test

5.Test For Bile salts – Hays sulphur test

6. Test For Bile pigments– Fouchets test

Report :- The Given Urine Sample Contains --------------------------- And

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