Denielle

 Genesis  B.  Camato  

   IV.  URINALYSIS  (LAB  TESTS/CHEMICAL  SCREENING)  
ANALYSIS  O F  URINALYSIS  AND  BODY  FLUIDS  |  REVIEWER  

COMPONENTS OF BASIC (ROUTINE) URINALYSIS

4 PARTS:
r Specimen evaluation
r Gross/ physical examination
r Chemical screening
r Sediment examination

SPECIMEN EVALUATION
r Proper labelling
r Proper specimen for requested test
r Proper receptacle (container)
r Storage conditions (time, temperature)
r Preservative NOTES!!!
r Visible signs of contamination UROCHROME – proportional to the metabolic rate and
r Transportation increased during starvation, fever, thyrotoxicosis
Normal concentrated urine may show sedimentary deposit if
URINE COMPOSITION allowed to stand after cooling from the body temp
ê Normal urine contains 90-95% water and about (phosphates & urates)
60 G/day of solid constituents which may be
organic or inorganic in nature VARIATIONS OF URINARY COLOR
A. PHYSIOLOGICAL VARIATIONS
ORGANIC constituents of urine S. No. Color Interpretation
S.No. Constituent Concentration
1 Dark yellow Concentrated urine – Mild
(G/day)
dehydration
1. Urea 25-30
Vitamin B complex
2. Uric acid 0.5-0.8
theraphy
3. Creatinine 1-1.8
4. Hippuric acid 0.7-0.8
2 Orange Drug induced
3 Pinkish Excessive beet root intake
INORGANIC constituents of urine
S.No. Constituent Concentration
(G/day) B. PATHOLOGICAL VARIATIONS
1. Chlorides 10-15 S. No. Color Interpretation
2. Sodium 3-5 1 DEEP jaundice
3. Potassium 2-2.5 YELLOW
4. Calcium 0.1-1-2
2 Reddish Haematuria
5. Phosphates 0.8-1.3
3 Brownish Hemoglobinuria,
6. Sulphates 1.0-1.2
myoglobinuria and
7. Ammonia 0.7-0.8
porphyrias
GROSS PHYSICAL EXAMINATION 4 Brown to Alkaptonuria
black
I. APPEARANCE 5 Cloudy Pus cells and bacteria in
infected cells
COLOR 6. Smoky Red blood cells
ê roughly indicates the degree of hydration and 7 Black Iron theraphy
urine concentration w/c correlates to urine 8. Pinkish brown Presence of urobilin –
specific gravity Hemolytic anemias
r UROCHROME (UROBILIN) 9. Milky white Chyluria (presence of fat
ê yellow pigment of the urine; globules)
r UROERYTHRIN
ê pink pigment; uric acid or urate crystals (brick
dust deposit); should not be confused with blood
r PALE COLOR
ê high fluid intake
r DARKER COLOR
ê decrease fluid intake
r NUBECULAE
ê small cloudy patches in normal urine; mucus
from urinary tract

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 Denielle  Genesis  B.  Camato  
   IV.  URINALYSIS  (LAB  TESTS/CHEMICAL  SCREENING)  
ANALYSIS  O F  URINALYSIS  AND  BODY  FLUIDS  |  REVIEWER  

II. VOLUME OF URINE VARIATIONS IN SPECIFIC GRAVITY OF URINE

ê Normal volume 800-2,500 mL/day with an average of 1500 a. LOW SPECIFIC GRAVITY 1.007 OR LESS
mL/day. ê Hyponuria
ê Approximately 500 mL/day is the minimum volume of urine ê Conditions:
needed in normal health to remove waste products û Compulsives polydipsia
ê The volume of urine is affected by – û Diabetes insipidus
þ Fluid intake û Glomerulonephritis
þ Fluid loss û Pyelonephritis
þ Type of diet
þ Cardio-vascular status b. HIGH SPECIFIC GRAVITY OF URINE 1.025 OR MORE
þ Renal functions ê Hypernuria
ê Conditions:
VARIATIONS IN VOLUME OF URINE EXCRETED û Severe dehydration
û nephrotic syndrome (due to proteinuria)
A) POLYURIA û Diabetes mellitus (due to glycosuria)
û Adrenal insufficiency (excess of sodium in urine)
ê Polyuria implies an increase volume of urine excreted û Congestive heart failure
per day; generally volume of urine exceeding 2,500 û Hepatic diseases
mL/day is termed as POLYURIA.
û Extra renal water losses (fever, vomiting and
ê Conditions causing POLYURIA diarrhea)
þ Diabetes mellitus
þ Diabetes insipidus c. FIXED SPECIFIC GRAVITY
þ Late stage of chronic glomerulonephritis ê (ISOTHENURIA)
þ Drug induced – diuretics ê Is seen in chronic renal failure
þ Alcohol ê Specific gravity of urine is based on tubular function
þ Compulsive polydipsia ê In the late stages of chronic renal failure, kidneys fail
to concentrate or dilute urine, which has a constant
specific gravity ranging between 1.008-1.012 (average 1.010)
B) OLIGURIA same as that of plasma.
ê Volume of urine less than 500mL/day is termed as
OLiGURIA MEASUREMENT OF SPGR
þ Conditons causing OLIGURIA ê The specific gravity is measured by URINOMETER
þ Fever ê The instrument floats in the urine
þ Diarrhea (loss of fluid from extra renal ê The calibration mark that corresponds to the surface level
sites) of urine is read
þ Severe edema ê It is calibrated at 15°C, temperature correction is done by
þ Acute nephritis adding 0.001 for every 3 degree above 15°C or subtracting
þ Early stage of acute glomerulonephritis 0.001 for every 3 degree below 15°C.
þ Cardiac failure and hypertension (reduced ê Proteinuria increases SPGR, 0.003 is subtracted for every
circulatory volume) G/L of urinary protein

C) ANURIA IV. ODOR OF URINE

ê Complete cessation of urine or volume of excreted ê Nomal odor of urine has an aromatic odor
urine less than in 100 mL/day ê Variations:
ê Conditions causing ANURIA û Ammoniacal Odor
þ Acute tubular necrosis þ on keeping sample for a long time
þ Blood transfusion reaction û Acetone like Odor
þ Surgical shock þ ketonuria such as Diabetic ketoacidosis or
þ Bilateral renal stones starvation
þ Sulphonamide theraphy û foul smell
þ due to bacterial infections
III. SPECIFIC GRAVITY
ê The specific gravity indicates the concentrating ability V. pH
of the kidneys. ê normal urine is acidic, pH ranges between 4.5-8.0 with a
ê In normal health the urinary specific gravity ranges mean of 6.0 in 24hrs.
between 1.016-1.025, the average being 1.020. the specific ê variations of urinary ph:
gravity is affected by:
þ Volume of urine excreted
þ The amount of solids present in the urine

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 Denielle  Genesis  B.  Camato  
   IV.  URINALYSIS  (LAB  TESTS/CHEMICAL  SCREENING)  
ANALYSIS  O F  URINALYSIS  AND  BODY  FLUIDS  |  REVIEWER  

a. ACIDIC URINE 3) TEST FOR URINARY PHOSPHATES

Physiologically, it is found after (Ammonium molybdate test)
ü A protein rich diet Principle:
ü Heavy exercise § upon warming with ammonium molybdate in
Pathologically, it is found in conditions of acidosis, such as the presence of nitric acid, inorganic
ü diabetic ketoacidosis, phosphates are precipitated as canary yellow
ü respiratory acidosis, and ammonium phosphor molybdate.
ü high fever (breakdown of tissue proteins) Procedure:
§ to 3mL of urine, add a few drops of conc.
Nitric acid and a pinch of ammonium
b. ALKALINE pH molybdate. Warm it. Observe the yellow color
Physiologically, it is found after of the precipitate
ü Heavy meals Interpretation:
ü Diet rich in citrus fruits § Increased urinary phosphates:
ü Excessive intake of milk and atacids } Osteomalacia, hyperparathyroidism, acidosis
Pathologically, it is found in § Decreased urinary phosphate:
ü Urinary tract infections } Nephritis, parathyroid hypofunction, pregnancy,
ü Conditions of alkalosis hereditary fructose intolerance and galactosemia.

4) TEST FOR URINARY CALCIUM

CHEMICAL EXAMINATION (Potassium oxalate test)
Principle:
ANALYSIS OF NORMAL CONSTITUENTS § with potassium oxalate in acidic medium, calcium is
precipitated as calcium oxalate.
A) INORGANIC CONSTITUENTS Procedure:
§ to 2ml of urine, add 5 drops of 1% acetic acid and 5ml of
1) TEST FOR URINARY CHLORIDES potassium oxalate. White precipitate of calcium is formed
(Silver nitrate test) Interpretation:
Principle: § increased urinary calcium:
§ Silver chloride is precipitated in the presence } hyperparathyroidism, hyperthyroidism,
of nitric acid and silver nitrate. Hypervitaminosis D, multiple myeloma, renal stones,
Procedure: renal tubular acidosis, steroids and diuretic theraphy
§ take 2mL of urine and add 0.5 mL of
concentrated nitric acid and 1mL of silver 5) TEST FOR AMMONIA
nitrate. A white precipitate of silver chloride Principle:
appears. § ammonia is evaporated when urine is made alkaline. Urinary
Interpretations: ammonia is derived from glutamine and other amino acids.
§ Increased urinary chlorides: Procedure:
} Polydipsia, use of diuretics and Addison’s disease. § to 5ml of urine add 2% sodium carbonate til the solution is
§ Decreased urinary chlorides: alkaline to litmus. Boil the solution. Place a piece of moistened
} Excessive sweating, fasting, diarrhea, excessive vomiting, red litmus paper at the mouth of the test tube. Note the
edema, diabetes insipidus, infections and adrenocortical change in color to blue due to evolution of ammonia.
hyper function (Cushing’s syndrome) Interpretation:
§ increased urinary ammonia:
2) TEST FOR URINARY SULPHATES } diabetic ketoacidosis, ingestion of acid forming foods,
(Barium chloride test) excessive water ingestion, urinary tract infections.
Principle: § decreased urinary ammonia:
§ urinary sulphate is precipitated as barium } alkalosis, nephritis
sulphate on reaction with barium chloride
solution.
Procedure:
§ take 3mL of urine and add 1mL of conc. HCl
and 2mL of 10% barium chloride. White
precipitate indicates the presence of
sulphates
Interpretation:
§ Increased urinary suplhate:
} Homocystinuria, melanuria, obstructive jaundice,
hepatocellular jaundice, cyanide poisoning and high protein
diet.
§ Decreased urinary sulphates:
} Are observed in conditions of renal functional
movement.

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 Denielle  Genesis  B.  Camato  
   IV.  URINALYSIS  (LAB  TESTS/CHEMICAL  SCREENING)  
ANALYSIS  O F  URINALYSIS  AND  BODY  FLUIDS  |  REVIEWER  

B) TEST FOR ORGANIC CONSTITUENTS III. TEST FOR URIC ACID

I. TEST FOR UREA a. PHOSPO TUNGSTIC ACID TEST

(Sodium hypobromite test) Principle:
Principle: § uric acid is a reducing agent in alkaline medium. It
§ when urea is treated with sodium hypobromite, reduces phospotungstic acid to tungsten blue
it decomposes to give nitrogen. Procedure:
Procedure: § take 2ml of urine, add a few drops of phospo
§ to 2ml of urine in a test tube, add 4-5 drops of tungstic acid reagent followed by a few frops
sodium hypobromite. Oserve the effervescence 20% sodium carbonate. Observe the appearance of blue
of nitrogen gas color

a. UREASE TEST FOR UREA b. BENEDICT’S TEST

PRINCIPLE: Principle:
§ soyabean powder contains the enzymes urease. This enzyme § uric acid is soluble alkali. The blue color is developed due to
under pH 7-8 and temperature 55C decomposes urea into the reduction ofphospo tungstic acid by uric acid.
ammonia and carbon dioxide which together form Procedure:
ammonium carbonate (alkaline component) which changes § to 2ml of urine, add a few drops of Benedict’s uric acid
the slightly acidic reaction (yellow color) to alkaline reaction reagent and add a pinch of anhydrous sodium carbonate and
(pink color). mix. A deep blue color indicates the presence of uric acid.
Interpretation:
b. BIURET TEST FOR UREA § increased urinary uric acid:
PRINCIPLE: } cancers, leukemia, administration of ACTH, Wlson’s
§ urea when heated decomposes with the liberation of disease
ammonia and the formation of biuret. Biuret is dissolved in § decreased urinary uric acid:
water and develops a violet color forming a complex with } purine free diet, gout
alkaline copper sulphate solution.
INTERPRETATION:
§ increased urinary urea:
} fever, diabetes mellitus, excess of adrenocortical TEST FOR ABNORMAL CONSTITUENTS OF URINE
activity
§ ABNORMAL URINE
§ decreased urinary urea:
§ Substances which are not present in easily detectable
} metabolic or respiratory acidosis, nephritis
amounts in urine of normal healthy individuals but are
present in urine under certain diseased condition are said to
be “Abnormal” or “pathological” constituents of urine.
ü Reducing sugar
II. TEST FOR CREATININE ü Ketone bodies
ü Proteins
a. JAFFE’S REACTION ü Blood
Principle: ü Bile salts
§ Creatinine reacts with picric acid in the alkaline ü Bile pigments
medium to form a redish colored complex of ü Urobilinogen
Creatinine picrate § These constituents are present in normal health but in very
Procedure: small amount and are not detected by less sensitive
§ take 5ml of urine and add an aqueous solution of laboratory methods.
picric acid. Make the mixture alkaline with NaOH § Under certain pathological conditions their concentration is
solution. A red color is produced. increased and these are get detected. The urine is said to be
‘Abnormal’, under such conditions

b. Nitroprusside test
Procedure:
§ to 5ml of urine add a few frops of sodium nitroprusside and
make the solution alkaline with sodium hydroxide (NaOH).
§ A ruby red color is formed that turns yellow.
§ This test is also called Wey’s test.

c. Nitroprusside – Acetic acid test (Salkowski test)

Procedure:
§ take 5ml of urine, add a few drops of sodium nitroprusside
and then make the solution alkaline with NaOH.a ruby red
color is formed that turns yellow. To the yellow precipitate,
add an excess of acetic acid and heat the solution.
A green color is obtained that turns blue upon standing.
Interpretation:
§ creatinuria
} creatinuria occurs in uncontrolled diabetes mellitus,
thyrotoxicosis, myasthenia gravis, starvation, infancy,
pregnancy,muscular disorders and in growing period.
§ Increased urinary Creatinine Æmuscular disorders
§ Decreased urinary Creatinine Ærenal failure

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 Denielle  Genesis  B.  Camato  
   IV.  URINALYSIS  (LAB  TESTS/CHEMICAL  SCREENING)  
ANALYSIS  O F  URINALYSIS  AND  BODY  FLUIDS  |  REVIEWER  

TEST FOR REDUCING SUGARS TEST FOR PROTEINS

1) BENEDICT’S TEST 1) HEAT COAGULATION TEST

Principle: Principle:
§ benedict’s reagent contains sodium carbonate, copper § albumin is coagulated when heated, which are precipitated at
sulphate and sodium citrate. In the alkaline medium provided the iso-electric point, when acetic acid is added.
by Sodium carbonate, the copper remains cupric hydroxide. Procedure:
Sodium citrate acts as a stabilizing agent to prevent § fill ¾ of the test tube with acidified urine (add few drops of
precipitation of cupric hydroxide. in alkaline medium, sugars dilute acetic acid) and heat the top half of it. Lower part
form enediols, cupric ion are reduced, and corresponding serves as a control. Note the appearance of turbidity.
sugar is oxidized to sugar acid. § Acidification is necessary because:
Procedure: m in alkaline medium heating may precipitate carbonates
§ take 5ml of benedict’s rgnt, add 8 drops of urine. Boil for and phosphates.
2min or keep it I the boiling water bath for 5mins. A light m False negative results may be obtained since the
green, yellow or brick red color is produced depending on proteins are coagulated by heat at a pH near isoelectric
concentration of urinary glucose. pH.
Observations: m In alkaline medium proteins may not be precipitated
§ benedict’s teast is a semi quantitative test. The color of the owing to the formation of alkaline meta proteins that
precipitate gives rough estimate of the reducing sugars are not precipitated upon heating.
present in the given samples.
m Green color – up to 0.5g% BENCE JONE’S PROTEINS
m Green precipitate – 0.5-1.0g% (+) § Are light chain immunoglobulins
m Yellow Precipitate - 0 1.0-1.5g% (++) § Excreted in urine of a patient suffering from multiple
m Orange precipitate – 1.5 -2.0g% (+++) myeloma
m Brick red precipitate - >2.0g% (++++) § These proteins precipitate between 40-60 degree centigrade
Interpretation: § Upon further heating, turbidity disappears to reappears on
§ Positive benedict’s test signifies Glycosuria cooling
m Glycosuria is a non-specific term. Any reducing sugar § These proteins redissolve on boiling unlike albumin
found in urine is denoted by glycosuria
§ Lactosuria – in lactose intolerance 2) SULPHOSALICYLIC ACID TEST
§ Galactosuria – in galactosemia Principle:
§ Fructosuria – in hereditary fructose intolerance § negatively charged supho salicylic acid neutralizes the positive
§ Pentosuria – in essential Pentosuria charge on proteins causing denaturation, and hence
precipitation of proteins.
2) FEHLING TEST Procedure:
§ Another reduction test § to 1ml of urine add 3 drops of 20% Sulphosalicylic acid. A
§ Contains KOH and Sodium potassium Tartrate in place of turbidity or precipitate indicates the presence of proteins
sodium carbonate nd sodium citrate in Benedict’s rgnt § Absence of cloudiness means absence of proteins.
§ No used any more, since it is less sensitive, less specific nd
the trong alkali causes caramelisation of the sugars present S. No. Observation Inference
in given sample. (approx. protein
conc.) mg/100ml
GLYCOSURIA
1 Barely visible turbidity 5
§ Causes of Glucosuria are: (Glucosuria and Glycosuria) are
2 Distinct turbidity 10-30
used synonymously)
a. renal glycosuria – pregnancy, hereditary, diseases of 3 Moderate turbidity 40-100
renal tubules, heavy metal poisoning 4 Heavy turbidity 200-500
b. diabetes mellitus 5 Heavy 500
c. alimentary glucosuria flocculent/precipitation
d. hyperthyroidism, hyperpituitarism and hyperadrenalism
e. stress, severe infections, increased intracranial
pressure
§ examples of non-carbohydrate substances which give a 3) HELLER’S NITRIC ACID RING TEST
positive Benedict’s reaction are: Principle:
a. Creatinine § conc. HNO3 causes denaturation and hence precipitation of
b. Ascorbic acid proteins
c. Glucoronates Procedures:
d. Drugs; salycylates, PAS and Isoniazid § take 3-5ml of conc. Nitric acid. Incline the tube and to it add
carefully, 2-4ml 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.
Interpretations:
§ insignificant amounts of proteins are excreted in urine in
normal health not exceeding 20-80mg/dl. This small amount is
not detectable by routine methods.
§ Under certain conditions, as much as 26G or more proteins
may be excreted per day in urine.
§ The most common type of proteinuria is albuminuria,; hence
proteinuria and albuminuria are used synonymously.

5  

 
 Denielle  Genesis  B.  Camato  
   IV.  URINALYSIS  (LAB  TESTS/CHEMICAL  SCREENING)  
ANALYSIS  O F  URINALYSIS  AND  BODY  FLUIDS  |  REVIEWER  

PROTEINURIA TEST FOR KETONE BODIES

§ When proteins appear in urine in detectable amounts. It
is called proteinuria. It can be casue by – 1) ROTHERA’S TEST
£ Glomerular permeability Principle:
£ Reduced tubular reabsorption § nitroprusside in alkaline reacts with a ketone group to for
£ Secretion of proteins purple ring. It is given by acetone and acetoacetate, but not
£ Conc. Of low molecular weight proteins in the by Beta hydroxyl butyric acid.
plasma Procedure:
§ saturate 5ml of urine with solid ammonium sulphate and add
§ PHYSIOLOGICAL 0.5ml of freshly prepared sodium nitroprusside (5%). Mix well
m Causes: and add liquor ammonia from the side of tube. A purple ring
û Violent exercise at the junction of the liquid indicates the presence of ketone
û Pregnancy bodies.
û Postural
û Alimentary 2) GERHARDT’S FERRIC CHLORIDE TEST
Principle:
û Exposure to cold
§ a purplish color is given by aceto acetate. On a boiling
§ PATHOLOGICAL acetoacetate is converted to acetone and does not give this
test positive. This test is only given by acetoacetate and not
a) pre renal
by beta hydroxyl butyric acid directly.
û severe dehydration
Procedure:
û heart disease
§ add 10% ferric chloride solution drop by drop to 5ml of urine
û ascites (due to increased intra-abdominal
in a test tube. If phosphates are present, precipitates of
pressure) ferric phosphates may form, that should be filtered off
û severe anemia and the ferric chloride is added. False positive Gerhardt’s
û fever test may be obtained with salicylic acid and salicylates.
û collagen diseases
û toxemia of pregnancy

b) renal
û all inflammatory, degenerative or
destructive diseases of kidney; the most
common ones are:
û nephrotic syndrome TEST FOR β- OH butyric acid
û pyelonephritis § No direct test for B-OH butyric acid
û acute & chronic glomerulonephritis § Indirect test is performed
û nephrosclerosis § Procedure: add a few drops of acetic acid to urine diluted 1:1
û tuberculosis of kidney with distilled water. Boil for a few minutes to remove
û renal failure acetone and aceto acetic acid. Add about 1.0ml of H2O warm
gently, cool, and perform Rothera’s test
c) post renal § Acetone, acetoacetate and beta hydroxy butyrate are
ketone bodies. Ketonemia and hence ketonuria occurs mostly
û also called false proteinuria because these
in conditions of glucose deprivation.
conditions proteins do not pass through the
kidneys. § Causes:
ü Uncontrolled diabetes mellitus
û Causes includes:
ü Starvation
û severe urinary tract infections
ü High fat feeding
û inflammatory, degenerative or traumatic
ü Heavy exercise
lesions of pelvis, ureters, bladder, prostate
ü Toxemia of pregnancy
or urethra
û bleeding genitor urinary tract
û pus in urine
û contamination of urine by semen or vaginal
secretions

6  

 
 Denielle  Genesis  B.  Camato  
   IV.  URINALYSIS  (LAB  TESTS/CHEMICAL  SCREENING)  
ANALYSIS  O F  URINALYSIS  AND  BODY  FLUIDS  |  REVIEWER  

TEST FOR BILE PIGMENTS TEST FOR UROBILINOGEN

1) FOUCHER’S TEST 1) EHRLICH’S TEST

Principle: Principle:
§ BaCl2 reacts with sulphate in urine to form barium sulphate. § the test for urobilinogen is based on the Ehrlich Aldehyde
If bilirubin is present I urine, It adheres to precipitate and is Reaction. P-dimethylaminobenzaldehyde in an acid medium
detected by oxidation to form biliverdin (green) with FeCl3 in with a color enhancer reacts with urobilinogen to form a
the presence of thrichloro acetic acid. Nitric acid oxidizes pink-red color. The optimum temperature for testing is 22-
bilirubin to biiverdin giving different colors from green to 26°C. a freshly voided sample is best for optimal results
violet. Procedure:
Procedure: § take 5ml of fresh urine in a test tube and add 5ml of
§ take 5ml of 10% BaCl2 to 10ml of urine and filter. Dry the Ehrlich’s rgnt to it. Wait for 10min and add 10ml of saturated
filter paper and add a few drops of Fouchet’s reagent sodium acetate solution. A pinkish color indicates the
(prepared by adding 10mg of 10% FeCl3 to 100ml of 25% TCA). A presence of urobilinogen. Porphobilinogen is also detected by
green color is obtained due to oxidation of bilirubin to Ehrlich’s test. The color intensifies upon addition of sodium
biliverdin. acetate of Porphobilinogen is there.
Interpretation:
IODINE TEST § urobilinogen is found in urine in hepatic and prehepatic
Procedure: jaundice. It its present in excessive amount in prehepatic
§ dilute some tincture of iodine with one to two volumes of jaundice and is completely absent in post hepatic jaundice. An
water and layer it carefully on to some urine in a test tube, increased urobilinogen conc. In urine is a sensitive index of
green ring at the junction of two fluids indicates the liver dysfunction or hemolytic disorders.
presence of Bilirubin. It is not a sensitive test, cannot detect
small amount of bilirubin present in the given sample.
Interpretation: TEST FOR BLOOD
§ bilirubin in urine means increased amount of conjugated
bilirubin because unconjugated bilirubin is water insoluble and is 1) BENZIDINE TEST
also bound to albumin, hence cannot cross the glomerular Principle:
membrane. § hydrogen peroxide liberated from Hb oxidizes Benzidine to
§ Causes: form a colored derivative
ü Moderate to severe hepatocellular damage Procedure:
ü Obstruction of bile duct – intra or extra hepatic § to 3ml of saturated Benzidine solution in glacial acetic acid,
ü In prehepatic jaundice, bilirubin is absent in urine. add 2ml of urine and add 1ml of 3% H2O2. A blue or green color
developswithin 10min. indicating the presence of blood. Color
developing after 10mins is not a positive test but it is due to
oxidation of Benzidine by atmospheric oxygen.
TEST FOR BILE SALTS Interpretation:
§ presence of blood in urine is called HEMATURIA.
1) HAY’S SULPHUR TEST m Gross hematuria
Principle: ê urine appears reddish in gross hematuria
§ bile salts lower the surface tension allowing the sulphur and this is observed in renal stones,
powder to sink malignancies, trauma, tuberculosis and
Procedure: acute glomerulonephritis.
§ sprinkle a little dry sulphur powder on the surface of fresh m Microscopic hematuria
urine in a test tueb taking distilled water as control. Sulphur ê blood is not visible to naked eye. It is
powder sinks in the presence of bile salts. In the control, observed in :
sulphur powder remains immiscible with the underlying liquid. ü Malignant hypertension
In the positive test, the sulphur sinks to the bottom ü Sickle anemia
Interpretation: ü Coagulation disorders
§ bile salts and bile pigments are present in urine in obstructive ü Polycystic kidney disease
jaundice. ü Incompatible kidney disease
ü Auto immune hemolytic anemia

URINE STRIP
ü 10 different substances in urine can be detected
ü Easy, quick and bedside procedure

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