URINE ROUTINE
ANALYSIS
Submitted to:
DEPARTMENT OF BIOCHEMISTRY
Dr. RPGMC TANDA, KANGRA (HP)
Submitted by: Ayushi, Jagriti, Mahesh, Sanjana, Smriti, Tanu
SPECIMEN COLLECTION
• FIRST MORNING SAMPLE- The morning sample is considered best as it
has higher specific gravity and lower ph and diserible for prevention of
formed element.
• Record collection time Use clean, dry sterile container.
• Analysed within 1hour of collection.
• Free of vaginal secretions.
PURPOSE OF URINE ANALYSIS
Urinalyses are performed for several reasons:-
• General evalution of health.
• Diagnosis of systemic diseases that affect kidney function.
• Diagnosis of endocrine disorders. Twenty-four-hour urine studies are often ordered
for these tests.
• Testing for pregnancy.
• Screening for drug anbuse.
COMPOSITION OF NORMAL URINE
• VOLUME- 600-2500ml/24hr (average-1200)
• SPECIFIC GRAVITY-1.003-1.030
• REACTION- acidic (Ph 4.7-7.5) Average Ph-6.0
• TOTAL SOLIDS- 30-70g/l
Inorganic constituents of Organic constituents
urine24/hrs of urine/24hrs
• IRON= 0.06-0.1mg • NITROGENOUS COMPOUND= 25-35gm
• CHLORIDE= 9-16gm • UREA= 15-30gm
• CREATINE= 60-150mg
• SODIUM= 3-4gm
• AMMONIA= 0.3-1.0g
• PHOSPHATE= 1.5- • URIC ACID = 0.3-1.0g
2.0g • PROTINE= 0-0.1gm
• SULPHER= 0.7-3.5gm
• CALCIUM= 0.1-0.3g
• All the specimans for routine urinalysis should be examined while fresh (within one
hour of collection).When urine is kept for longer than one hr before analysis,to avoide
deterioratin of chemical and cellular material and prevent multiplication of bacteria, it
should be stored at 2-8*c in a refrigerator.
The expected changes in the composition of urine stored at room temperature
are as follow:-
Lysis of red blood cells by hypotonic urine.
Decomposition of casts.
Bacterial multiplication.
Decrease in glucose level, due to bacterial growth
TYPES OF ANALYSIS
• Physical Examinaation
• Chemical Analysis(urine dipstick or manual method)
• Microscopic Examination
PHYSICAL ANALYSIS
It Includes
• VOLUME
• COLOR
• APPERANCE
• ODOUR
• VOLUME: for adults normal average daily volume of urine is about 1200-
1500ml. The normal range of 24hr urine may be form 600-2000ml.
• COLOR: The color of normal urine may vary from pale yellow to dark amber.
Very pale or colourless urine can result from high fluid consumption, dirutic drugs,
natural dirutic such as alcohol and coffee and also in clinical condition such as
diabetes insipidus and diabetes mellitus.
• APPEARENCE: normal urine is usually clear. Urine may appear cloudy or
turbid from the presence of leucocytes and epithelial cells and confirmed by
microscopic examination. Bacteria can also cause cloudiness to urine fat and chyle
give urine a milky color. Presence of RBCs may give urine turbid and smokey
appereance.
• ODOUR: presence of ketone bodies gives urine a sweet or fruity smell. A
contaminated urine with bacteria may give pungent smell due to the formation of
ammonia.
CHEMICAL ANALYSIS
The routine urinalysis includes chemical testing for-
• Protein
• Glucose
• Ketone bodies
• Occult blood
• Bile pigments
• Bile salts
• Urobilinogen
TEST FOR PROTEIN
HEAT AND ACETIC TESTS
Principle
• Heat induced coagulation of proteins and precipitation. Coagulation can further enhanced
when drops of acetic acid are added.
PROCEDURE
Fill three-fourth ½ of a test tube with clear urine.
Heat the upper part (1/3) of the urine.
(The lower part of the urine acts as a control)
Add a few drops of (3-5) 10% glacial acetic acid
INTERPRETATION
• If turbidity persists, it is due to proteins.
• Depending upon the amount of precipitate, results are interpreted below.......
TEST FOR GLUCOSE
BENEDICT’S TEST
PRINCIPLE:
The copper sulphate present in Benedict's reagent reacts with the reducing substances in
the urine which convert cupric sulphates to cuprous oxide in hot alkaline media.
PROCEDURE
Take 5 mL of Benedict's qualitative reagent in a test tube
Boil to exclude the presence of reducing substance in reagent
Add 8 drops (0.5 mL) of protein-free urine.
Boil the mixture for 5 min.
Allow to cool (under running tapwater)
INTERPRETATION
• The ratio of 5 mL of Benedict's reagent and 8 drops (0.5 mL) of urine ratio is important
because it is a Semi-quantitative test.
• Interpretation:
The change of color from blue to green, yellow orange / red depends on the amount
of sugar present.
TEST FOR KETONE BODIES
ROTHERA’S TEST
PRINCIPLE:
Acetoacetic acid and acetone react with sodium nitroprusside in the presence of an alkali
to form a purple color compound.
PROCEDURE
Take 4 mL of urine in a test tube
Add a few crystals of sodium nitroprusside
saturate the urine with ammonium sulphate By mixing
vigorously
Overlay with a few drops of liquor ammonia along the wall of
the tube
INTERPRETATION
• Development of purple ring indicates the presence of Acetoacetic acid/acetone or both.
• A brown or red color is of no significance.
TEST FOR BILE SALTS
HAY’S SULFUR TEST
PRINCIPLE:
Bile salts have unusual property of lowering the surface tension of urine markedly even
when present in small concentrations.
PROCEDURE
Take 10 mL urine in a wide bore test tube or small beaker
Sprinkle sulfur powder over its surface, watch for 5 min
INTERPRETATION
• Sulfur powder sinks to the bottom of the test tube in the presence of bile salts in the
urine
• CAUSES:
Hepatocellular and obstructive Jaundice
MICROSCOPIC EXAMINATION OF
URINE
• REQUIREMENTS:
• centrifuge or test tube
• Glass slides
• Coverslip
• Pasture pipettes centrifuge
• Microscope
• Speciman- fresh ueine sample
PROCEDURE
• Mix the urine and pour into a centrifuge tube until it is 3/4 th fill
• Centrifuge with another balanced test tube for 5min at 2500rpm
• Discard the supernatant and mix the sediment
• Place on drop of sediment on glass slide
• Cover with coverslip
• Observe under microscope
NORMAL REPORT
ABNORMAL REPORT
OBSERVATIONS
• ORGANISED ELEMENT
RBC’s
WBC’s
Epithelial cells
Casts-Hyaline
Cellular
Granular
Waxy and fatty casts
ERYTHROCYTES
In fresh urine these cell have a normal
pale yellow appereance.
It appear smooth biconcave disks
They do not contain nuclei.
LEUCOCYTES
Normal urine contains 2-3 pus cells.
These are mostly neutrophils.
EPITHELIAL CELLS
Normally few cells 3-5 from
these site found in urine
Three types of epithelial cells
1. Tubular
2. Transitonal
3. Squamous
1. TUBULAR EPITHELIAL
These are slightly larger than leucocytes
and contain large round nucleus
They may be cuboidal, flat or columnar
2. TRANSITIONAL
EPITHELIAL
They may be pear shaped or round.
These cell may contain 2 nuclei
3. SQUAMOUS
EPITHELIAL
These are flat large and irregular in shape
They contain small central nuclei
CASTS
• Urinary castes are formed in the lumen of the tubule of the kidney.
• Casts dissolve in alkaline urine .
• These are of following types:
a) Granular casts
b) Hyaline
c) Red cell casts
d) White cell casts
e) Epithelial cell casts
f) Waxy casts
g) Fatty casts
a) GRANULAR CASTS: These always b) HYALINE CASTS: They are
indicate significant renal diseases. The colorless, homogeneous,
casts are present due to the degeneration transparent with rounded nucleus.
of cellular casts.
c.) RED CELL CASTS: The cast may d.) WHITE CALL CASTS: that
contain only a few RBCs in protein appear in casts and
matrix. Presence of red cell always polymorphonuclear neutrophils. They
pathogenic. are tightly packed together.
e.) EPITHILIAL CALL CASTS: f.) WAXY CASTS: They have high
The epithelial cells may be arranged refrective index. These are yellow grey or
haphazardly and vary in size and shape. colorless and have a homogeneous
appereance.
g.) FATTY CASTS:
• These are formed by incorporated free fat droplets or oval fat bodies.
CRYSTALS
• CALCIUM OXALATE CRYSTALS: These are colorless and envelop shaped. They
also appear as oval, sphere or biconcave disks which have dumbbell shape.
• AMORPHOUS URATES: These are urate salts of sodium, potassium, magnesium and
calcium.
• SODIUM URATE: These may be present as amorphous or or as crystal. These are
colorless or yellowish needle in clusters.
• CALCIUM SULFATE CRYSTALS: These are long thin and colorless needle or
prisms.
• CYSTINE CRYSTALS: These are colorless hexagonal plates with equal an
unequal sites.
• TYROSIN: These appear in the form of fine refractile needles, occurring in crystals.
• CHOLESTEROL: These crystals are large flat and in the form of transparent plateswith
notched corners.
