QUALITATIVE EXAMINATION FOR
NORMAL AND PATHOLOGIC ORGANIC
CONSTITUENTS OF URINE
Raralio, Ranier| Sandoval, Maria Tricia Anne C. |
Joseph | Uy, Pauline | Villarba, Jan Margaret P.| Wepee,
Ralph Ryan
3BIO6
URINE
Excretory product of the body
Removes excess water and water soluble
wastes the kidney filters from the blood
Reflects the work of the kidney to
maintain normal pH homeostasis
An aqueous solution that ranges from
colorless to amber (dark orange-yellow)
Diuresis
COMPOSITION OF URINE
FORMATION OF URINE:
1.GLOMERULAR FILTRATION
Process by which the kidneys filter the
blood , removing excess wastes and fluids
Glomerular filtration rate (GFR) is a
calculation that determines how well the
blood is filtered by the kidneys, which is
one way to measure remaining kidney
function
INTRODUCTION
Tolentino,
COMPOSITION OF URINE
Formation of urine:
1. Glomerular filtration
2. Tubular reabsorption
3. Tubular secretion
COMPOSTION OF URINE
FORMATION OF URINE:
2. TUBULAR REABSORPTION
Process by which solutes and water are
removed from the tubular fluid and
transported in to the blood
Reabsorption is a two-step process
beginning with the active or passive
extraction of substances from the tubule
fluid into the renal interstitium, and then
the transport of these substances from the
interstitium into the bloodstream
Mainly driven by active transport
 COMPOSITION OF URINE COMPOSITION OF URINE
FORMATION OF URINE: 95% water
THE OTHER 5%
3. TUBULAR SECRETION 9.3g/L urea
Transfer of materials from peritubular capilliaries 1.87 g/L chloride
to renal tubular luman 1.17g sodium
Secretion is caused mainly by active transport 0.750 g/L potassium
Few substances are secreted 0.670 g/L creatinine
Present in great excess or are natural poisons OTHER COMPONENTS:
Magnesium, calcium, ammonium sulfates, phosphorus,
hormones, fatty acids, pigments, enzymes and
mucins

URINE OUTPUT
URINALYSIS
NORMAL URINE OUTPUT: 1- 2 Liters Routine urine test
ABNORMAL URINE OUTPUT:
OLIGURIA
Evaluates a sample of urine and to assess and
detect disorders
-Less than 500 mL of urine in the course of 24 hours
ANURIA Urine Examinations:
-Less than 100 mL of urine in the course of 24 hours Physical Initial Examination
POLYURIA Examines color, volume, turbidity, odor, and pH
-More than 2.5 L or urine in the course of 24 hours
Qualitative Examination
Detection for the presence of normal organic and
pathologic constituents

URINALYSIS OBJECTIVES
To make an initial examination of the
urine sample
Helps detect substances in the urine To make a qualitative examination for
associated with different metabolic and normal organic constituents of the
kidney disorders urine sample
To make a qualitative examination for
pathologic organic constituents of the
urine sample side by side with a
positive control

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 A. Initial Examination

Urine sample

METHODOLOGY

Indicate the time of collection.
Note the color of the urine (indicate whether it is light or dark)
Note the clearness or turbidity (check for suspended
materials/sediments)
Check the pH using pH paper or meter (indicate whether the
sample is acidic, neutral or basic)

Examined urine
sample
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Test for Urea

B. Qualitative Examination 1mL Urine
for Organic Constituents Sample

Add 0.5mL 70% NaOH

Add 4 drops bromine water

Bubble formation
(N2 gas)

Test for Uric Acid Test for Creatinine

1mL Urine Sample 2mL Urine Sample

Add 5mL of 20% Na2CO3

Add 1mL of alkaline picrate
Add 5 drops of solution (5:1, saturated
phosphotungstic acid reagent picric acid-10% NaOH)

Blue solution Orange solution

 Indican Test
C. Qualitative Examination
for Pathologic Organic
5mL Urine Sample
Constituents
Add 5mL of Obermayers reagent
Add 3mL chloroform
Shake and allow the chloroform layer to
settle.

Blue color in lower

layer

Gunnings Test ( for Ketone Bodies) Benedict's Test (for Glucose)

5mL Urine Sample / 8-10 drops Urine

Positive control: 5mL Sample / Positive
urine + 2mL acetone control: 1mL urine + 1%
glucose solution
Basify with 5 drops concentrated
Add 5mL of Benedicts reagent
ammonium hydroxide
Heat in a boiling water bath for 2-3 minutes
Add Lugols solution
Let stand and allow to cool.
Let stand for 5 minutes.

Iodoform Crystals Precipitate

Smiths Test (for Bile Pigments) Extons Test (for Albumin)

5mL Urine Sample / 3mL Urine Sample /
Positive control: 1mL Positive Control: 3mL
urine + 3-5 drops bile urine + pinch of
pigment from chicken Albumin
or pig gall bladder
Incline the test tube Add 3mL of Extons reagent.
Overlay with 3mL of tincture of Warm the solution. *If
alcoholic iodine mixture. cloudiness appears

Emerald green
solution Cloudy Solution
 Test for Occult Blood
3mL acidified Urine Sample
(with acetic acid) / Positive
control: 3mL acidified urine + 3
drops of blood RESULTS AND
Add 5mL of 95% ethanol to a half
spatula guaiac
Add 5mL of hydrogen peroxide.
DISCUSSION
Add 5mL of this solution to acidified
urine sample/positive control.

Blue ring formation

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A. INITIAL EXAMINATION OF URINE

INITIAL EXAMINATION OF URINE
GROUP Time of Color of Clear/Turbi PH LEVEL
NUMBER collection Urine d GROUP #6 NA NA NA pH 7
sample
GROUP #1 Midmorning Yellow- NA NA GROUP #7 Midmorning Light yellow CLEAR pH 7 and 8
(9:55 AM) orange (slightly
basic)
GROUP #2 Morning Light Slightly pH 6 GROUP #8 Midmorning Clear CLEAR pH 5 (Acidic)
(9:15 AM) yellow turbid yellowish
GROUP #3 Midmorning Lightly Clear pH 6 (Acidic) GROUP #9 Midmorning Dark yellow CLEAR pH 5 (Acidic)
colored (10:50 AM)
GROUP #4 First Light Clear pH 6 (Acidic)
morning yellow GROUP #10 Midmorning Bright CLEAR pH 5 (Acidic)
specimen (10:40 AM) yellow
(5AM)
GROUP #5 Midmorning Light NA pH 7 Light yellow NA pH 6 (Acidic)
GROUP #11
( 10:30 AM) yellow (Neutral)
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Test for Urea

Groups Results
Group 1 Formation of bubbles
Group 2 Evolution of bubble and
gas
Group 3 Solution with bubbles

B. Examination for Normal Organic Group 4 Clear solution with bubbles

Group 5 Accumulation of bubbles
Constituents of Urine Sample
Group 6 Light yellow turbid solution
with bubbles
Group 7 Formation of bubbles
Group 8 Formation of bubbles
Group 9 Presence of heat observed;
bubbles observed
Group 10 Heat produced; With
Bubbles
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Group 11 Light Yellow solution with
bubbles on top
 Test for Creatinine
Test for Uric Acid
Groups Results Groups Results

Group 1 Blue solution Group 1 Orange colored solution

Group 2 Clear, faint blue Group 2 Orange solution

solution Group 3 Turbid red-orange solution

Group 3 Clear, faint blue solution
Group 4 Blue color formation on Group 4 Orange colored solution

top of clear colorless Group 5 Clear red-orange solution

solution Group 6 Clear red solution with orange
Group 5 Clear light blue solution precipitate
Group 7 Orange clear solution
Group 6 Clear light blue solution
Group 8 Red-orange colored solution
Group 7 Clear blue solution
Group 8 Clear blue solution Group 9 Red-orange solution with
Group 9 Clear light blue solution small sediments present
Group 10 Bright clear orange solution
Group 10 Clear light blue solution
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Group 11 Clear light blue solution
Group 11 Clear red-orange solution

Indican Test
Group 7 Lower layer: clear light
Group 1 Sand-like yellow lower
blue solution
layer
Group 2 Clear lower layer Group 8 Clear orange solution
with blue chloroform
Group 3 Faint blue color on layer
chloroform layer Group 9 Upper layer: Brown clear
Group 4 Upper layer: Yellow-orange solution; bubbles present
solution in between and the lower
Lower layer: Clear colorless layer is very light blue.
solution
Upper Layer: Brown
Group 5 Formation of blue color in Group 10 solution with bubbles
the chloroform layer Light blue solution at the
Group 6 Upper layer: Clear brown bottom
solution Group 11 Upper layer: Gold yellow
Middle layer: Clear yellow solution
solution Lower layer: Light blue
Lower layer: Clear light solution
blue solution

Gunnings Test
Groups Normal Urine Sample Positive Control
Group 1 Yellow solution with Light turbid yellow
black precipitate at solution with white
the bottom crystals
Group 2 Black precipitate; clear Dark brownish red
C. QUALITATIVE EXAMINATION solution solution
Group 3 Clear yellow solution Clear yellow solution
FOR PATHOLOGIC ORGANIC with yellow and black with yellow
CONSTITUENTS precipitate precipitate
Group 4 Light yellow solution Colorless solution
with black precipitate with light yellow
at the bottom and precipitate
crystals
Group 5 Yellow solution Light yellow
without white solution with white
crystals crystals
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 Group 6 Clear dark brown Clear yellow solution Benedicts Test
solution with brown with white precipitate
precipiate Groups Normal Urine Sample Positive Control
Slightly turbid light Formation of yellow
Group 1 Clear blue solution Green solution with
Group 7 yellow solution with dark crystals with cloudy
with white white precipitate
green precipitate substances suspended
crystals in slightly turbid light precipitate
yellow solution Group 2 Clear blue solution Clear yellow
Group 8 Yellow turbid solution; Yellow clear solution solution
no crystals with crystals Group 3 Clear blue solution Turbid yellow
Group 9 Orange clear solution Slightly turbid light with yellow solution with
with white crystals at yellow solution; white precipitate yellow precipitate
the bottom crystals settled at the Group 4 Clear blue solution Turbid muddy
bottom green solution with
Group 10 Formation of white Formation of orange precipitate
crystals at the bottom precipitate (cloud like)
with white crystal Group 5 Turbid light yellow Turbid light
precipitate above blue solution yellow precipitate
Group 11 Turbid colorless Turbid yellow solution underneath blue
solution with black
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color solution
precipitate

Group 6 Slightly turbid blue White slightly turbid

solution with yellow solution with white Extons Test
precipitate precipitate
Group 7 Bright blue clear solution Chrome yellow Groups Normal Urine Sample Positive Control
with no precipitate precipitate with light
green turbid solution Group 1 Clear light yellow solution Turbid/cloudy yellowish
white solution
Group 8 Blue clear solution; no Yellow-green solution
precipitate formed with orange precipitate
Group 2 Clear solution Clear solution
Group 9 Upper layer: Blue turbid Light green and turbid
solution with sediments solution; brownish Group 3 Clear slightly yellow Cloudy solution with
afloat yellow colored solution white solid
Lower layer: clear blue precipitate that settled
solution at the bottom
Overtime the sediments Group 4 Clear light blue solution Turbid/Cloudy light blue
clumped up with bubbles solution

Group 10 Blue solution with yellow Solution turned into

precipitate turbid green solution
Group 5 Clear light yellow solution Turbid white solution
with yellow precipitate with solid particles
underneath
Group 11 Clear Blue green solution 11/25/2015
Turbid yellow green 11/25/2015
solution with yellow
precipitate

Group 6 Clear light brown solution Cloudy turbid solution

Smiths Test
Group 7 Rosepink clear solution Slightly turbid rosepink
solution with cloudy
suspensions and Groups Normal Urine Sample Positive Control
precipitate Group 1 Upper layer: Red solution Pig Bile: Without
Lower layer: Yellow emerald green at point
solution of contact
Group 8 Yellowish solution Cloudy solution No emerald green color at Chicken Bile: Emerald
Group 9 Slightly turbid colorless White precipitate formed point of contact green at point of contact
solution at the solution
Group 2 No emerald green; clear Emerald green lower
lower layer layer
Group 10 Yellow solution at the top Presence of cloudy solution
Group 3 Green interphase; Clear Upper layer: Reddish
and clear colorless solution
solution below brown solution
at the bottom
Lower layer: Green
solution; with green
Group 11 Clear colorless solution Turbid white solution with
interphase
white cloud like
precipitate

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 Group 7 Yellow color at point of Light blue color at point
Group 4 Upper layer: dark red solution Upper layer: Dark red solution
Lower layer: Clear colorless Lower layer: Clear yellow contact of contact
solution solution; Emerald green at Upper layer: dark brown Upper Layer: Dark
point of contact and red clear solution brown and red clear
Lower layer: clear yellow solution
solution Lower layer: Clear
yellow solution
Group 8 Clear colorless solution; no Yellow-green turbid
Group 5 Upper layer: dark brown Upper layer: dark brown
solution solution emerald green color solution with green
Lower layer: clear colorless Lower layer: green solution interphase
solution Emerald green at the point of
No emerald green at the point contact Group 9 Upper layer: Dark red Upper layer: dark red
of contact thick solution thick solution
Group 6 Clear solution upon contact Chicken: Clear solution upon Lower layer: Very light Lower layer: slightly
with white precipitate and a contact with white
brown upper layer precipitate
yellow clear solution turbid emerald green
Pig: Emerald solution upon solution
contact with white
precipitate

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Test for Occult Blood

Groups Normal Urine Sample Positive Control
Group 1 Dark brown solution with Light brown turbid
brown precipitate and solution and absence of
absence blue ring blue ring
Group 10 No presence of emerald Emerald green Group 2 No blue ring, turbid Brown ring; Turbid
brown solution with black brown solution with
green interphase and clear interphase; Turbid
precipitate black precipitate
colorless solution at the yellow solution at the
Group 3 Turbid brown solution with Upper layer: Yellow
bottom bottom
dark brown precipitate turbid solution
Lower layer: Blue
solution; No ring formed
Group 4 Upper layer: Dark brown Upper layer: Dark brown
solution solution
Group 11 Clear colorless Emerald green Lower layer: Clear Lower layer: Turbid
solution; no emerald interphase; colorless solution solution
green color Upper layer: Dark With light brown Presence of blue ring and
red solution precipitate with dark brown
Lower layer: Light precipitate
green solution Group 5 Clear light yellow Turbid white solution
solution with solid particles
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underneath
Group 6 Clear light brown Cloudy turbid
solution solution

Group 7 Light brown turbid Upper layer: Turbid

solution light brown solution
Lower layer: presence
of bluish gray
solution
Group 8 Turbid peach solution; Turbid peach solution;
no blue ring no blue ring
PHYSICAL
Group 9
Dark brown colored at
the top; light brown
Blue ring is present;
Upper layer: dark
brown solution
PROPERTIES OF
solution Lower layer: light
brown precipitate at URINE
the bottom of the
solution
Group 10 Brown turbid solution Presence of blue layer
near the bottom
Group 11 Upper layer: Dark Upper layer: Brown
brown solution turbid solution with
Lower layer: Light black precipitate
brown solution with
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Lower layer: Blue 11/25/2015

brown precipitate turbid solution

 Types of Urine Specimens Types of Urine Specimens

Random specimen 2-Hour postprandial specimen

may be collected at unspecified times
suitable for most screening purposes.
First morning specimen or 8-hour specimen
to collect the specimen immediately upon rising from a
nights sleep
to void shortly before consuming a routine meal
and to collect a specimen 2 hours after eating.
24-hour (or timed) specimen
obtain an accurately timed specimen

Fasting specimen Catheterized specimen

second voided specimen after a period of fasting. collected under sterile conditions by passing a
hollow tube through the urethra into the bladder

Types of Urine Specimens Types of Urine Specimens

Midstream clean catch specimen

First morning specimen or 8-hour
provides a safer, less traumatic method for obtaining urine
for bacterial culture.
specimen is valuable because it is more
concentrated and abnormalities are
Suprapubic aspiration easier to detect.
Urine may be collected by external introduction of a needle
into the bladder. Early morning specimen is relatively
Pediatric specimens free of dietary influences.
may be collected by attaching a soft, clear plastic bag with
adhesive to the general area of both boys and girls.

Physical Examinations of Urine

COLOR
Colorless to Amber(dark orange to yellow) normal color of
urine (due to the presence of urobilin,
uroerythrin,and urochromes)
Varies because of recent diet intake
Urochrome pigment that contributes to the color
of the normal urine
Discolorations of Urine:
1. Colorless high water intake or chronic renal failure
2. Yellow Orange/Orange dehydration or due to vitamins and supplements
3. Red/Brown bleeding in the urinary tract
4. Cloudy urinary tract infections
 GROUP Time of Color of Clear/Turbi PH LEVEL

COLOR INTERPRETATION NUMBER collection Urine

sample
d

GROUP #1 Midmorning Yellow- NA NA

(9:55 AM) orange
Yellow/orange: concentrated urine, urobilinogen,
bilirubin GROUP #2 Morning Light yellow Slightly pH 6
(9:15 AM) turbid
Red/red brown: RBC, hemoglobin, myoglobin, GROUP #3 Midmorning Lightly Clear pH 6 (Acidic)
menstrual contamination, porphyrin, amidazophen colored
Brown black/black: melanin, methemoglobin, GROUP #4 First Light yellow Clear pH 6 (Acidic)
morning
homogentisic acid specimen
Blue-green: pseudomonas infection, chlorophyll, (5AM)
biliverdin, methylene blue GROUP #5 Midmorning Light yellow NA pH 7
( 10:30 AM) (Neutral)
Milky/cloudy: lipids, mucus, dye, microorganisms
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GROUP #6 NA NA NA pH 7

GROUP #7 Midmorning Light yellow CLEAR pH 7 and 8

(slightly
Physical Examinations of Urine
basic) ODOR
GROUP #8 Midmorning Clear CLEAR pH 5 (Acidic)
yellowish Aromatic Odor - normal
GROUP #9 Midmorning Dark yellow CLEAR pH 5 (Acidic) Pungent/Ammonia Smell due to urinary tract
(10:50 AM) infections, dehydration, or high-protein foods or
simply bacteria
GROUP #10 Midmorning Bright CLEAR pH 5 (Acidic)
Fruity Smell due to ketonuria
(10:40 AM) yellow
Ammoniacal Odor due to hydrolysis of urea to ammonia
GROUP #11 Light yellow NA pH 6 (Acidic) May provide health information
For example, urine of diabetics may have a sweet or fruity odor due
to the presence of ketones
Fresh urine has a mild smell
11/25/2015 Aged urine has a stronger odor similar to that of ammonia.

Physical Examinations of Urine

pH LEVEL Alkaline pH
The pH of normal urine ranges from 4.5 8.2 Physiologically found after
Average pH level = 6.0 - heavy meals
Variations of urinary pH - diet rich in citrus fruits
Acidic Urine - excessive intake of milk and
Physiologically found after
antacids
- a protein rich diet
Pathologically
- high exercise
- urinary tract infection
Pathologically
- conditions of acidosis such as diabetic
- alkalosis
ketoacidosis, respiratoy acidosis and high fever
 GROUP Time of Color of Clear/Turbi PH LEVEL GROUP #6 NA NA NA pH 7
NUMBER collection Urine d
sample GROUP #7 Midmorning Light yellow CLEAR pH 7 and 8
GROUP #1 Midmorning Yellow- NA NA (slightly
(9:55 AM) orange basic)
GROUP #8 Midmorning Clear CLEAR pH 5 (Acidic)
GROUP #2 Morning Light Slightly pH 6 yellowish
(9:15 AM) yellow turbid Midmorning Dark yellow CLEAR pH 5 (Acidic)
GROUP #9
GROUP #3 Midmorning Lightly Clear pH 6 (Acidic) (10:50 AM)
colored
GROUP #4 First Light Clear pH 6 (Acidic) GROUP #10 Midmorning Bright CLEAR pH 5 (Acidic)
morning yellow (10:40 AM) yellow
specimen
(5AM) GROUP #11 Light yellow NA pH 6 (Acidic)
GROUP #5 Midmorning Light NA pH 7
( 10:30 AM) yellow (Neutral)

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Physical Examinations of Urine

Physical Examinations of Urine
DENSITY VOLUME
Normal Urine Output: 1- 2 Liters
Density determines the urines overall concentration
Abnormal Urine Output:
Normal urine ranges from 0.001 to 0.035
Oliguria
Dark urine = higher density; light urine = lower
-Less than 500 mL of urine in the course of 24 hours
density
Anuria
-Less than 100 mL of urine in the course of 24 hours
Polyuria
-More than 2.5 L or urine in the course of 24 hours

Physical Examinations of Urine

Physical Examinations of Urine
SPECIFIC GRAVITY
A measurement of the kidneys ability TURBIDITY
to concentrate and dilute urine
High specific gravity indicates Clear solution Normal
concentrated urine. Low specific
gravity indicates dilute urine. caused by the separation of a cloud
Specific Gravity of Urine: 1.000 - 1.035 called nebecula which is composed of
Usual Specific Gravity of Urine: mucoid and epithelial cells
1.010- 1.025
 GROUP Time of Color of Clear/Turbi PH LEVEL
Causes of Turbidity:
NUMBER collection Urine d
Amorphous Phosphate and sample
Carbonates GROUP #1 Midmorning Yellow- NA NA
Crystals, Cellular Exudates, (9:55 AM) orange
Bacteria and Fungi
GROUP #2 Morning Light Slightly pH 6
Chyle and Fat (9:15 AM) yellow turbid
Pus GROUP #3 Midmorning Lightly Clear pH 6 (Acidic)
colored
Amorphous Urates in Acidic
GROUP #4 First Light Clear pH 6 (Acidic)
Urine
morning yellow
specimen
(5AM)
GROUP #5 Midmorning Light NA pH 7
( 10:30 AM) yellow (Neutral)

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GROUP #6 NA NA NA pH 7

GROUP #7 Midmorning Light yellow CLEAR pH 7 and 8

(slightly Qualitative Examinations of Urine
basic)
GROUP #8 Midmorning Clear CLEAR pH 5 (Acidic) Tests for Pathological
yellowish Tests for Normal Organic
Organic Constituents:
Midmorning Dark yellow CLEAR pH 5 (Acidic) Constituents:
GROUP #9 Gunning's Test
(10:50 AM) Test for Urea
Benedict's Test
GROUP #10 Midmorning Bright CLEAR pH 5 (Acidic) Test for Uric Acid Exton's Test
(10:40 AM) yellow Test for Creatinine Smith's Test
Light yellow NA pH 6 (Acidic)
Test for Occult Blood
GROUP #11

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Test for Urea

Qualitative
Examination for Normal
Organic Constituents
Urea (CH N O) 4 2 Urea
Major end product of protein nitrogen metabolism in
mammals
It is produced in the liver and excreted
Hepatic enzymes convert ammonia from amino acids to through the kidneys in the urine.
urea.
Levels of urea depend on protein intake,
Ammonia (NH3) protein catabolism and kidney function
Product of oxidative deamination reaction Normal urea level in the urine is 12 to 20
Toxic even in small amounts grams over 24 hours.

Urea Urea
Elevated levels of urea can occur because of:
dietary changes
Urea Cycle:
diseases which impair kidney
function Also known as Ornithine cycle
liver diseases
Cycle of biochemical reactions
congestive heart failure
Diabetes
occurring in many animals that converts

infection. ammonia and carbon dioxide to urea in

the liver

Test for Urea

Sodium hypobromite - when reacted with urea releases a brisk
effervescence (Nitrogen Gas)

Reagents: 0.5mL 70% NaOH, Bromine water

Principle: NaOH hydrolyzes urea. Excess NaOH reacts with bromine

water to form NaOBr which oxidizes urea to form N2,CO2, H2O

Positive result: bubble formation (N2 gas)

 Results
Test for Urea Groups
Test for Urea
Results
Group 1 Formation of bubbles
Group 2 Formation of bubbles
Mechanism Group 3 Solution with bubbles
Group 4 Clear solution with bubbles
Group 5 Accumulation of bubbles
Group 6 Light yellow turbid solution
with bubbles
Group 7 Formation of bubbles
Group 8 Formation of bubbles
Group 9 Presence of heat observed;
bubbles observed
Group 10 Heat produced; With Bubbles
Group 11 Light Yellow solution with
bubbles on top

Test for Urea Test for Urea

Indications:

Normal: 12 to 20 grams in 24 hours High urea level:

Low urea level: Fever, diabetes mellitus, excess adrenocortical

Liver diseases, nephritis, metabolic or respiratory activity, heart failure, dehydration, too much protein

acidosis, kidney problem, not enough protein intake intake

Test for Uric Acid Uric Acid

is an heterocyclic compound of carbon, nitrogen,
oxygen and hydrogen.

Chemical formula: C5H4N4O3

Is the waste product of purine catabolism.

Slightly soluble in water

Easily precipitates out of solution forming needle-

like crystals of Sodium Urate
Uric Acid
Contribute to the formation of kidney
stones
High levels of uric acid can produce
excruciating pain ofgoutor kidney disease.
Normal values range between 3.5 and 7.2
mg/dL.
Test for Uric Acid
Indications:
Normal range of uric acid in a 24 hour period: 250 750 mg
Hyperuricemia (greater-than-normal levels
of uric acid) may be due to:
Acidosis , Alcoholism
Chemotherapy-related side effects
Diabetes, Renal failure, Lead poisoning
Purine-rich diet

Test for Uric Acid Test for Uric Acid

Lower-than-normal levels of Reagents: 20% Na2CO3, Phosphotungstic Acid

Principle: Uric acid is oxidized by

uric acid may be due to:
phosphotungstic acid to allantoin. The
Fanconi syndrome
reduction product of phosphotungstate in
Low purine diet
Syndrome of inappropriate antidiuretic alkaline medium is tungsten blue.
hormone (SIADH) secretion
Positive result: Blue colored solution
Wilson's disease

Test for Uric Acid

Mechanism

Indican TestAlso known as Obermeyer test

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 Indican Indican

Normally, only a small amount of indican

Is an indole produced by bacterial action on
amino acid, tryptophan, in the intestine is found in the urine.

Most of the indole is excreted in the feces. The amount of urine indican increases
Remainer is absorbed, metabolized and with high protein diets or inefficient
excreted as indicant in the urine.
protein digestion.

Indican Indican
If protein is not digested adequately, bacteria act on
Indole (oxidized) indoxyl +
the protein causing putrefaction in the colon and the
H2O2 indoxyl sulfuric acid K + indoxyl
production of indoles, which are absorbed and
potassium sulfate (indican)
converted in the liver to Indican.
Formation of Indican:
Excess urine Indican may be caused by:
Maldigestion and/or malabsorption of protein
Bacterial overgrowth in the small and/or large
intestine

Indican Test Indican Test

Mechanism
Reagents: Obermeyer s reagent (Ferric chloride dissolved
in fuming hydrochloric acid), Chloroform

Principle: Hydrochloric acid will hydrolyze indoxyl sulfate;

ferric chloride accelerates the oxidation of indoxyl to
indigo. Chloroform enhances the visibility of indigo.

Positive result: Blue color in the lower (chloroform) layer

Indican Test
Detection of indican in the urine depends upon its
decomposition and subsequent oxidation of indoxyl to
indigo blue and its absorption into a chloroform layer.
Test for Creatinine
Your Result
Negative (Normal) = Clear or blue tinge
1+ (Slightly = Slight blue, yellow,
Positive) mint green
2+ (Positive) = Dark blue, light
green, golden brown
3+ (High Positive) = Violet, indigo,
dark brown
98

Creatinine Test Creatinine

Also known as Jaffes reaction

Creatinine Creatinine is formed from creatine

By-product of muscle metabolism Creatine is synthesised in the liver and
kidneys
Amount increases with muscle
mass 3 amino acids involved in creatine synthesis
Formed from creatine and Arginine, Glycine, and Methionine
phosphocreatinine Enzymes involved
Excreted in the urine Arginine: Glycine amidinotransferase
Guanidinoacetate N-methyltransferase

99 100

Creatinine Test Creatinine Test

Reagents:
Alkaline Picrate solution (5:1, saturated picric acid-10%
NaOH)
Principle:
Reaction of Creatinine with Picrate ions in the alkaline
medium forming a orange to reddish complex of
Creatinine and Picric Acid
Positive result:
Orange to Red Orange coloured solution

101 102
 Creatinine Test Creatinine Test
Mechanism
Indications
Normal Creatinine levels depend on muscle
mass
To check: if kidneys are working normally, if
kidney disease is changing, how well kidneys
work in people who drink medications that
can cause kidney damage, severe dehydration

103 104

Gunnings test
For Ketone Bodies

Qualitative Examination
for Pathological Organic
Constituents

Test for Ketone

Test for Ketone Reagents:
Conc. NH4OH, Lugols solution (Iodine and KI dissolved
Molecules that are produced by the liver from in distilled water)
fatty acids when not enough carbohydrates are
available for the cells to use Principle:
Conc. NH4OH will basify the urine it will give the
High levels of ketone bodies are toxic enolate of acetone. If acetone is present, iodoform
crystals will appear. Black cloud precipitation of
Little or no ketone bodies must be present in the iodine-nitrogen.
urine
Positive result:
Acetoacetate and Beta-hydroxybutyric acid are Iodoform crystals
commonly found in urine
Test for Ketone
Positive Result

110

Groups Normal Urine Sample Positive Control

Group 6 Clear dark brown solution Clear yellow solution

Group 1 Yellow solution with black Light turbid yellow solution with brown precipiate with white precipitate
precipitate at the bottom with white crystals Slightly turbid light yellow Formation of yellow
Group 7 solution with dark green crystals with cloudy
precipitate crystals substances suspended in
Group 2 Black precipitate; clear Dark brownish red solution slightly turbid light
solution yellow solution

Group 3 Clear yellow solution with Clear yellow solution with Group 8 Yellow turbid solution; no Yellow clear solution with
yellow and black precipitate yellow precipitate crystals crystals
Group 9 Orange clear solution with Slightly turbid light
white crystals at the bottom yellow solution; white
Group 4 crystals settled at the
Light yellow solution Colorless solution with light
yellow precipitate bottom
with black precipitate at
Group 10 Formation of white crystals Formation of precipitate
the bottom and crystals at the bottom (cloud like) with white
crystal precipitate
Group 11 Turbid colorless solution Turbid yellow solution
Group 5 Yellow solution without Light yellow solution with with black precipitate with white precipitate
white crystals white crystals

Test for Ketone

Benedicts Test
Normal amount of ketone in urine is 3-15 mg daily
Test for Sugar
High levels of ketone in the urine
Diabetic ketoacidosis, starvation, low carbohydrate diet,
Anorexia.

Ketonuria
Ketone is present in urine
Glucose Test for Glucose
Reagents:
Benedicts reagent (Sodium citrate + sodium carbonate
Digested food that became usable source of dissolved in water with heating + copper sulfate dissolved
energy in water)
0-0.8 mmol/l (0-15 mg/dL
Principle:
Reduction of copper sulfate to form cuprous oxide in an
alkaline medium in the presence of heat
Sodium carbonate: makes the medium alkaline
Sodium citrate: acts as the stabilizing agent to prevent
deterioration of Cupric ions
Positive result:
Brick red precipitate

Test for Glucose Test for Glucose

Color results
Range
No color change (Blue) = sugar is absent

Green = trace amounts of sugar

Yellow = Low amounts of sugar

Orange = Moderate amount of sugar

Brick Red = Large Amount of sugar

117

Benedicts Test
Test for Glucose Groups Normal Urine Sample Positive Control

Range: Blue = sugar is absent, green = 0.5%, yellow = 1%,

Group 1 Clear blue solution with white Green solution with white
orange = 1.5%, brick red = 2%+
precipitate precipitate

Group 2 Clear blue solution Clear yellow solution

Group 3 Clear blue solution with yellow Turbid yellow solution with
precipitate yellow precipitate

Group 4 Clear blue solution Turbid muddy green solution

with orange precipitate

Group 5 Turbid light yellow above blue Turbid light yellow precipitate
solution underneath blue color solution
 Slightly turbid blue solution with White slightly turbid solution with
Test for Glucose
Group 6
yellow precipitate white precipitate

Group 7 Bright blue clear solution with no Chrome yellow precipitate with light
precipitate green turbid solution
Indications
Group 8 Blue clear solution; no precipitate Yellow-green solution with orange
formed precipitate
Group 9 Upper layer: Blue turbid solution with Light green and turbid solution;
Normal glucose range in urine: 0 - 0.8
sediments afloat brownish yellow colored precipitate mmol/l (0 - 15 mg/dL)
Lower layer: clear blue solution that settled at the bottom
Overtime the sediments clumped up
High levels of glucose in urine
Diabetes, Renal glycosuria, pregnancy

Group 10 Blue solution with yellow precipitate Solution turned into turbid green Glycosuria
solution with yellow precipitate
presence of glucose in urine
Group 11 Clear Blue green solution Turbid yellow green solution with
yellow precipitate

Albumin

Extons Test Protein made by the liver

Naturally found in the plasma of the blood
(for albumin) Soluble in water
Precipitated by acid
Coagulated by heat

Extons Test Groups Normal Urine Sample Positive Control

Group 1 Clear light yellow solution Turbid/cloudy yellowish

Reagents: Extons reagent ( Na2 SO4 and white solution

sulfosalicylic acid)
Principle: The Na2SO4 and sulfosalicylic acid acidify Group 2 Clear solution Clear solution
the urine causing the protein to precipitate making Group 3 Clear slightly yellow Cloudy solution with white
the solution cloudy. The heat coagulates albumin. solution solid

Positive result: Cloudy solution Group 4 Clear light blue solution Turbid/Cloudy light blue
with bubbles solution

Group 5 Clear light yellow solution Turbid white solution with

solid particles underneath

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 Extons Test
Group 6 Clear light brown solution Cloudy turbid solution INDICATIONS:
Group 7 Rosepink clear solution Slightly turbid rosepink
solution with cloudy
suspensions and Albuminuria- the presence of albumin in urine
precipitate
Increased presence of albumin means:
Group 8 Yellowish solution Cloudy solution kidney damage, diabetes and pregnancy
Group 9 Slightly turbid colorless White precipitate formed
solution at the solution Normal amount of albumin in urine is less
Group 10 Yellow solution at the top Presence of cloudy solution than 30mg
and clear colorless solution
at the bottom

Group 11 Clear colorless solution Turbid white solution with

white cloud like
precipitate
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Bile Pigments

Smiths test Naturally occurring compound

Metabolic product of certainporphyrins
(for Bile Pigments)
Jaundice- caused by the accumulation of
biliverdin or bilirubin (or both) in the
circulatory system and tissues

Smiths Test
Reagents: Tincture of alcoholic iodine ( iodine crystals
Biliverdin- a green tetrapyrrolicbile and sodium iodide dissolved in absolute ethanol)
pigment, and is a product ofheme Principle: Iodine serves as the oxidizing agent. Bilirubin
catabolism. Pigment responsible for the is oxidized to biliverdin
greenish color seen in bruises
Positive result: Emerald green
Bilirubin- a yellow pigment in bile and is
responsible for the yellow color of Bile pigments in the urine indicates:
bruisesand the yellow discoloration Jaundice, liver diseases
injaundice.
Bile- a fluid that is made and released by
the liver and stored in the gallbladder.
 Group 7 Yellow color at point of Light blue color at point
contact of contact
Upper layer: dark brown and Upper Layer: Dark brown
Smiths Test red clear solution and red clear solution
Lower layer: clear yellow Lower layer: Clear yellow
solution solution
Groups Normal Urine Sample Positive Control
Group 1 Upper layer: Red solution Pig Bile: Without emerald
Group 8 Clear colorless solution; no Yellow-green turbid
Lower layer: Yellow solution green at point of contact
No emerald green color at Chicken Bile: Emerald green emerald green color solution with green
point of contact at point of contact interphase
Group 9 Upper layer: Dark red thick Upper layer: dark red
solution thick solution
Group 2 No emerald green; clear Emerald green lower layer
lower layer Lower layer: Very light Lower layer: slightly
Group 3 Green interphase; Clear Upper layer: Reddish brown yellow clear solution turbid emerald green
solution below solution solution
Lower layer: Green solution; Group 10 No presence of emerald Emerald green interphase;
with green interphase green interphase and clear Turbid yellow solution at
colorless solution at the
the bottom
bottom

Group 11 Clear colorless solution; no Emerald green interphase;

emerald green color Upper layer: Dark red
solution
Lower layer: Light green
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solution

Occult Blood/ Hematuria

Blood that cannot be seen without a

microscope.
Can be caused by bladder or kidney
Test for Occult Blood infections or stones, kidney diseases,
enlarged prostate, sickle cell anemia, and
other urogenital infections

Test for Occult Blood Test for Occult Blood

Reagents: Guaiac powder in 95% Ethanol & Hydrogen

Peroxide Presence of occult blood indicates:
Principle: Hemoglobins peroxidase activity Kidney infection or stones, urogenital
decomposes hydrogen peroxide; the liberated oxygen infections and even bladder infection
oxidizes the guaiac powder forming the blue ring.
Positive result: Blue ring

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 Group 5 Solution without blue Solution with blue ring,
Test for Occult Blood
ring, with solid with solid particles
particles underneath; underneath; Dark brown
Light brown solution on solution on top
Groups Normal Urine Sample Positive Control top

Group 1 Dark brown solution with Light brown turbid

brown precipitate and solution and absence of Group 6 Brown turbid solution Brown turbid solution
absence blue ring blue ring with brown precipitate with gray precipitate
Group 7 Light brown turbid Upper layer: Turbid
Group 2 No blue ring, turbid Brown ring; Turbid brown solution light brown solution
brown solution with black solution with black Lower layer: presence
precipitate precipitate of bluish gray solution
Group 3 Turbid brown solution Upper layer: Yellow
with dark brown turbid solution Group 8 Turbid peach solution; Turbid peach solution;
precipitate Lower layer: Blue no blue ring no blue ring
solution; No ring formed
Group 9 Blue ring is present;
Group 4 Upper layer: Dark brown Upper layer: Dark brown Dark brown colored at Upper layer: dark
solution solution the top; light brown brown solution
Lower layer: Clear Lower layer: Turbid solution Lower layer: light
colorless solution solution brown precipitate at
With light brown Presence of blue ring and the bottom of the
precipitate with dark brown solution
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precipitate

CONCLUSION
Group 10 Brown turbid Presence of blue
solution layer near the
bottom
An initial example of urine was conducted
Group 11 Upper layer: Upper layer: and the time, color, pH level and haziness
Dark brown Brown turbid otherwise known as the physical
solution solution with properties of urine were noted.
Lower layer: black
Light brown precipitate Qualitative examinations for normal
solution with Lower layer: organic constituents was made: Urea,
brown Blue turbid indican, and creatinine were present
precipitate solution among the tested urine samples.

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REFERENCES
Croftan, A.C. (1904). Clinical Urinology William Wood & Company:
USA.

Qualitative examination for Pathologic

Organic constituents was made: 4 groups Hammarsten, O. (2013). pp. 558-9.A Text-Book of Physiological
Chemistry. London: Forgotten Books.
tested positive for the presence of ketone
(Gunninge test), 1 group tested positive
Marakala, V., Avinash S., Shivashankara A., et al. (2012) Serum
for the presence of albumin while Bile Creatinine assay: Enzymatic Vs Kinetic Jaffe's method Srinivas
(Smiths Test) and Kidney stones (Occult Institute of Medical Science & Research Centre: Mangalore

blood Test) cannot be determined in the

urine samples presented. McClatchey, K. (2002). Clinical Laboratory Medicine Lippincott
Williams & Wilkins: USA.

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Mundt, L., Shanahan, K. (2011). Graff's Textbook of
Routine Urinalysis and Body Fluids 2nd Ed Lippincott
Williams & Wilkins, Wolters Kluwer business: USA.

Weatherby, D. Ferguson, S. (2003). In-Office Lab

Testing: Functional Terrain Analysis 2nd Ed Bear
Mountain Publishing: USA.

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