METABOLISM and

ENERGY PRODUCTION
ANALYSIS OF URINE

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 URINE
a sterile liquid byproduct of the body
secreted by the kidneys, and
excreted through the urethra
through a process called urination

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CHEMICAL COMPONENTS
IN URINE
▸ Water ▸ Calcium
▸ Urea ▸ Magnesium
▸ Sodium ▸ Leukocytes
▸ Chloride ▸ Nitrates
▸ Potassium ▸ Urobilinogen
▸ Sulfate
▸ Protein
▸ Creatinine
▸ Ammonia
▸ Glucose

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1.
COLOR, pH, &
SPECIFIC GRAVITY

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UROCHROME

▸ Yellow pigment that imparts

a yellow color to urine
▸ Came from the byproducts of
rbc destruction

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 pH test of urine

▸ Measures the acidity of the urine which may

indicate sign of kidney stones
▸ Urine pH varies during the day
▹ AM : Acidic
▹ PM : BASIC / ALKALINE
▸ Normal urine pH : 4.6-8
▸ ADVANTAGES OF TESTING THE URINE pH:
▹ helps in determining kidney stones or
kidney failure
▹ Helps in determining if urinary tract
infection exist
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 URINE REAGENT STRIP (pH)
▸ color ranges from red-orange to yellow
then yellow-green to blue-green
▸ values in this test can only distinguish
pH values ranges from 5-9

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 SPECIFIC GRAVITY
▸ ratio of urine to water density
▸ normal urine : 1.001 to 1.030
▸ To gauge a patient’s hydration
status as well as the functional
ability of his kidneys

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URINOMETER AND REFRACTOMETER
URINOMETER REFRACTOMETER
▸ an instrument used ▸ determines the amount of
to measure specific dissolved solids in liquid
gravity of urine solution by passing light
▸ based on the through sample and show
principle of refracted angle
Buoyancy ▸ the more dissolved solids
water contains = the slower
light travels, and more
bending effect

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2.
PROTEIN

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Proteinuria

▸ Occurs when too much protein is

excreted in the urine.
▸ Excreted protein should not exceed 150
mg per day.

Protein is too large to pass through the

glomeruli

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Prolonged foamy urine 5 minutes ( A ) and 15 minutes ( B ) after urinating
directly into a beaker. Normal pattern (not foamy) urine after remission of the
proteinuria ( C ).
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 Types of Proteinuria
Pre-renal Renal Proteinuria: Post-renal
Proteinuria: Includes all forms of Proteinuria:
Caused by functional and Due to protein that
overflow/overload pathologic is deposited in the
proteinuria. proteinuria. urine from the upper
Presence of urinary tract,
hemoglobin, bladder, prostate or
myoglobin, and urethra.
Bence-Jones
proteins.

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 Types of Renal Proteinuria
Functional Renal Proteinuria
▸Transient: may be
caused by fever or
exercise
▸Orthostatic: change of
body position
Pathologic Renal Proteinuria
▸Glomerular: increased
permeability of glomerulus
▸Tubular: failure to
reabsorb small proteins in
the tubules
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Protein Error of Indicators

▸ The ability of protein to alter the color

of some acid-base indicators without
altering the pH.
▹ Dipstick test for Protein and
Microalbumin

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 Protein Error of Indicators
False Positive: False Negative:
▸ Highly buffered or ▸ Primary protein is
alkaline urine not albumin (e.g.
▸ Bacterial Bence-Jones
contamination, protein)
alkaline ▸ In the presence of
medication, or curcuma or
skin cleansers w/ turmeric
chlorhexidine

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Microalbuminuria

▸ Refers to small detectable amounts of

albumin in the urine (30-300 mg/day).
▸ One of the first signs of renal disease or
damage that can lead to renal failure.
▸ Associated with diabetes, Chronic
Kidney Disease, cardiovascular disease,
and hypertension.

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Microalbuminuria
Reagent Strip Tests
▸ The test to detect microalbuminuria is
done by reagent strips.
▸ It is based on dye binding using
sulfonephthalein dye.
▸ At constant pH, albumin binds with
sulfonephthalein dye to develop a blue
color.

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3.
GLUCOSE

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Glycosuria

▸ Occurs when high amounts of glucose

is found in the urine.
▸ When there are high amounts of glucose
in the blood, the kidneys may not be
able to absorb back all of them from the
urine.

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Renal Threshold for Glucose

▸ Renal Threshold is the plasma

glucose concentration above
which significant glycosuria
occurs
▸ Normal threshold: 160-180 mg/dL

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Glucose Oxidase method
of reagent strip test

This test detects concentrations of 100 to

2000 mg/dL
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Glucose Oxidase method
of reagent strip test

False Positive: False Negative:

▸ Presence of ▸ Presence of
oxidizing ascorbic acid and
detergents and bacteria
peroxide
contaminants

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4.
KETONE BODIES

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 KETONE BODIES

▸ Ketone bodies are produced by the liver and used

peripherally as an energy source when glucose is
not readily available.

▸ Ketones are a byproduct of fat being metabolized

and used as a source of energy.

▸ Ketones are not usually detected in urine but if

there is high amount of ketone in urine one might
have KETONURIA. 26
 KETONURIA
▸ Medical condition in which ketone bodies are present in urine.

▸ 3 types of Ketone bodies present in urine

1. Acetoacetate

2. Beta-hydroxybutyric acid

3. Acetone

▸ Causes of Ketonuria

1. Metabolic abnormalities

2. Dietary conditions

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 KETONE BODIES : LEGAL’S TEST

▸ Determines the presence of ketone

bodies.

▸ Reagent : Sodium Nitroprusside and

glycine.

▸ Positive Result : Purple colored

complex.
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Principle of Nitroprusside reaction,
sensitivity and possible causes of
interference
▸ Acetoacetic acid in an alkaline medium will react with nitroprusside and
glycine to produce a purple color.

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Principle of Nitroprusside
reaction, sensitivity and
possible causes of interference

False-Positive False-Negative
● Phthalein
compounds that
produce red
● Improperly
colouration.
preserved
● Highly
specimens.
pigmented red
urine.
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 Dipstick test in detecting
presence of ketone bodies

▸ This test is done to detect the

presence of ketone bodies but
with the use of reagent sticks.
▸ Presence of ketone bodies will
cause a reaction and change in
color in the pads of the sticks.

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Similarities in the dipstick test
and legal’s test in detecting
ketone

▸ Both tests have a violet colored

result as their positive result.
▸ Sodium nitroprusside act as their
reagents.

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5.
UROBILINOGEN &
BILE PIGMENTS

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OUTLINE OF RBC TO UROBILIN

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 INTERPRETATION OF ABNORMAL
RESULT OF UROBILINOGEN

INCREASE DECREASE
▸ liver disease ▸ Reduce Bile Flow
▸ Urobilinogen ▹ Cholestasis
overload ▸ Certain
caused by Medications
release of larger ▸ Exposure of
quantities of urine sample to
hemoglobin direct sunlight
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 RELATIONSHIP OF URINARY BILIRUBIN
AND UROBILINOGEN WITH THE
DIAGNOSED BILIARY OBSTRUCTION, LIVER
DISEASE, AND HEMOLYTIC DISEASE

URINE BILIRUBIN UROBILINOGEN

BILE DUCT
POSITIVE (HIGH) Negative
OBSTRUCTION

POSITIVE OR
LIVER DAMAGE POSITIVE
NEGATIVE

HEMOLYTIC POSITIVE OR
POSITIVE
DISEASE NEGATIVE
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 REAGENT STRIP TEST (BILIRUBIN)
▸ based on the coupling of bilirubin with diazotized
dichloroaniline in a strongly acid medium
▸ color range from light tan to reddish-brown
▸ POSSIBLE SOURCES OF ERROR IN GETTING THE
BILIRUBIN AMOUNT:
▹ Urine is not fresh
▹ Large dose of ascorbic acid

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6.
BLOOD

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 CLINICAL SIGNIFICANCE

HEMATURIA HEMOGLOBINURIA MYOGLOBINURIA

- Presence of blood - Presence of free - Presence of

in urine hemoglobin in myoglobin in urine
- 2 TYPES: Gross and urine - Rhabdomyolysis or
Microscopic - Hemolytic anemia muscle
- Renal or genital destruction
disorders

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 APPEARANCE
● Urine color
HEMATURIA HEMOGLOBINURIA MYOGLOBINURIA

- Red, pink, or dark - Red to amber - Red to amber

brown

● After centrifugation
HEMATURIA HEMOGLOBINURIA MYOGLOBINURIA

- Clear supernatant - Pigmented - Pigmented

- Sediment red supernatant supernatant
- Red plasma - Clear plasma

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GUAIAC TEST
- Occult blood test
- RESULT: Green to blue colored complex

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7.
ELECTROLYTES

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Sodium ion : Flame Test

▸ Flame test are used to detect the presence of small amounts of

metal ions.

▸ Flame colors are produced from the excitement of electrons.

▸ It is important that the wire should first be dipped in acid and

placed into the burner for several times before conducting the
test.

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Different flame colors
depending on the metal ion

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 Chloride ion: Silver nitrate test
▸ A test to determine the presence of chloride ions in a
specimen.

▸ Reagent: Silver nitrate or Nitric Acid.

▸ Positive result : cloudy, White precipitate.

▸ The Presence of albumin while conducting the test

might give a false positive result/

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Chemical Equation Involved in
the Silver Nitrate test

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8.
DIPSTICK

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 DIPSTICK TEST
- Uses a thin plastic
strip treated with
chemicals and is
placed in the
urine. It’s dipped
into your urine,
and the chemicals
on the stick react
and change color
if levels are above
normal.

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REAGENT STRIPS

- Made up of specific reagents which are

reactive with specific biological
material and produce very specific
observations to confirm the presence
and quantify the biological material.

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URINE COMPONENTS
DETECTED BY DIPSTICK
▸ pH: 4.6-8 (Low)
▸ Specific Gravity: 1.001-1.030 (Low)
▸ Protein: Negative
▸ Glucose: Negative
▸ Ketones: Negative
▸ Bilirubin: Negative
▸ Nitrite: Negative
▸ Leukocyte: Negative
▸ Blood: Negative
▸ Urobilinogen: Less than 17 µmol/l (<
1mg/dl)

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4 CAUSES OF PREMATURE
DETERIORATION OF REAGENT
STRIPS
▸ Moisture
▸ Volatile chemicals
▸ Heat
▸ Light

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5 QUALITY CONTROL PROCEDURES
1. Test open bottles of reagent strips with
known positive and negative controls
every 24 hours
2. Resolve control results that are out of
range by further testing
3. Test reagent used in backup test with
positive and negative controls
4. Perform positive and negative controls
on new reagents and newly opened
bottle of reagent strips
5. Record all control results and reagent
lot numbers
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BENEDICT’S TEST
- Test for reducing sugars
- REAGENT: Benedict’s reagent
- REACTION: Reduction
- RESULT: Brick red precipitate

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BIURET TEST
- General test for compounds (proteins and
peptides) having two or more peptide (CO-NH)
bonds.
- REAGENTS: Potassium hydroxide and Copper
sulfate
- REACTION: Complexation
- RESULT: Violet solution

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LEGAL’S TEST

- To detect ketones
- Based on nitroprusside reaction
- RESULT: Violet colored complex

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GMELIN TEST
- Test for the presence of bile pigments
- REACTION: Oxidation

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GUAIAC TEST

- Occult blood test

- RESULT: Green to blue colored complex

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THANK YOU FOR LISTENING!

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References

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