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UrinalysisJorge Anton D. Ordas
Eloisah Vin S. RagodonLindley C. Susi
Mercer Keith Von PosselMark Rainier C. Yu
3BIO6
Urine Is a composite of various organic and
inorganic substances which are normally waste products from metabolic processes.
Body fluids filtered by the kidney and excreted through the urethra
Typically yellow-amber in color Used in testing for physiological
imbalances or disease
Urine (composition) Average Urine
Output(Adults): 1-2 liters per 24 hours
5% - solutes 95% - water Main Organic
Constituents: Urea, Creatinine, Uric acids, Ammonium, SO4 /Sulfate, and PO4.
Main Ions present: Sodium, Potassium, Chloride, Magnesium and Calcium ions
URINE (Output/Volume) Oliguria
Less than 400 ml of urine in 24 hours
Anuria Less than 100 ml of urine in 24 hours
Possible causes:1. Prerenal – dehydration, congestive heart failure and
hemorrhage2. Postrenal – obstruction of the urinary tract 3. Renal parenchymal disease - acute tubular necrosis,
chronic renal failure
Urine (increase) Polyuria
More than 2L of urine in 24 hours
Possible causes: 1. Physiological states: increased water intake/
excessive hydration, some drugs/ diuretic effects, intravenous solutions
2. Pathologic states: Diabetes mellitus, Diabetes insipidus
Urinalysis Diagnostic tool used for the overall
evaluation of renal function or possible malfunction/disease
Rapid detection and measurement of various compounds that pass through the urine
Includes 2 steps of examination: Physical Characterization– color, turbidity, presence
of suspended particles Chemical Reaction Tests– detection of normal and
pathologic organic constituents
Urinalysis (Importance)
To identify or detect the presence if excess protein, glucose, bile pigments are left unfiltered by the renal tubules, intact or hemolysed RBCs and elevated level of intestinal microbiota and possible etiologic agent of urinary tract infection and renal diseases
Objectives To make an initial examination of the
urine sample
To examine the normal organic constituents of the urine sample
To examine for pathologic organic constituents of the urine sample by comparing with positive control set-ups
Procedure/Methodology
A. Initial Examination of Urine• Note time of collection• Indicate the color and the turbidity and
check for suspended materials• Check pH through litmus paper
Procedure/Methodology
B. Examination for Normal Organic Constituents
B1. Test for urea
1 mL urine
sample
Observe formation of
bubbles
-add 0.5 mL 70% NaOH and 4 drops bromine water
Procedure/Methodology
B2. Test for uric acid1 mL Urine
Sample
Record Result
-add 5 mL of 20% Na2CO3
, then mix-add 5 drops phosphotungstic acid reagent, then mix-observe for formation of blue solution
Procedure/Methodology
B.3 Indican test5 mL Urine
Sample
Record Result
- add 5 mL Obermayer’s Reagent, then mix well
- add 3 mL chloroform. Shake and allow chloroform to settle.- observe for formation of blue color in lower layer
5 mL Urine Sample
Record Result
Procedure
B4. Creatinine test
2 mL Urine Sample
Record Result
-add 1 mL of alkaline picrate solution
-note for formation of an orange solution
ProcedureC. Examination of pathologic organic constituents
C1. Gunning’s test
Record Result
-Basify with 5 drops concentrated ammonium hydroxide .Check pH by red litmus paper.- add Lugol’s solution to sample until black cloud appears- note the formation of iodoform crystals
Positive Control5 mL Urine + 2 mL
acetone
5 mL Urine Sample
Procedure
C.2. Benedict’s test
Record Result
- add 5 mL of Benedict’s reagent - heat in boiling water bath for 2-3 minutes then cool - note precipitate formation
Positive Control1 mL Urine + 1 mL glucose solution
0.5 mL Urine Sample
Procedure
C3. Exton’s test
Record Result
-add 3 mL of Exton’s reagent-warm if cloudiness appears-if cloudiness persist or increase in heating, albumin is present
Positive Control3 mL Urine + pinch
of albumin
3 mL Urine Sample
Procedure
C4. Smith’s test
Note if emerald green is seen at point of contact
-incline test tube and overlay with 3 mL of tincture of alcoholic iodine mixture
Positive Control3 mL Urine + 3-5
drops bile pigment
5 mL Urine Sample
Procedure
C5. Test for occult blood
Record Result
- add 5 mL of 95% ethanol to half spatula guaiac powder then add 5 mL of hydrogen peroxide in one test tube
- note the formation of iodoform crystals
Positive Control3 mL acidified
Urine + 3 drops of blood
3 mL Acidified Urine Sample
- add 5 mL of this solution to 3 mL acidified urine and positive conrol
Results
Time collected
Color Turbidity pH
19/20/12 7:15
amDark yellow Turbid 6
2 9/20/12 7:30 am
Yellow Orange Clear 5
3 9/20/12 9:00 am
Light Pale Yellow
Slightly Turbid 8
4 9/19/12 10:59 pm
Light Yellow Slightly turbid w/ minute suspended particles
6
5 9/20/12 6:00 am
Dark Yellow Clear 6
6 9/19/12 8:25 pm
Dark Yellow Clear 6
7 9/20/12 7:45 am
Light Yellow Clear 6
8 9/20/12 10:00 am
Yellow Orange Turbid 6
9 9/20/12 9:55 am
Very light yellow
Clear 6
10 9/20/12 9:20 am
Light Yellow Clear 7
11 9/20/12 9:30 am
Golden Yellow Clear 6
A. Initial Examination of Urine Sample
Group Test for Urea
1White gas was observed then brown gas almost
immediately
2Clear yellow solution upper layer and clear light yellow
lower layer
3Light yellow solution, with N2
gases, presence of cloudy white substance
4 Formation of bubbles, light yellow solution
5 Formation of bubbles on top
6Clear yellow solution with formation of bubbles on the
uppermost portion
7Clear yellow solution with presence of bubbles and foamy
layer
8Light yellow color upper layer with white fumes and clear
yellow second layer
9Yellow solution with presence of bubbles indicating
formation of N2 gas
10 Evolution of gas was observed
11There was an evolution of N2
gas
B. Examination for Normal Organic Constituents of Urine Sample
Group Test for Uric Acid
1 Clear blue solution
2 Clear light blue solution
3 light blue colored solution appeared
4 Clear blue solution
5 Formation of blue clear solution
6 Clear blue solution
7 Clear light blue solution
8 Blue solution formed
9 Light blue solution
10 Light blue colored solution
11 Clear blue solution
Group Indican Test
1 Clear blue colored lower layer and dark green upper layer
2Clear brownish green upper layer and blue lower layer with
presence of emulsion
3Olive green solution at upper layer and pale clear blue
solution at lower layer
4Lower light blue clear solution, middle olive green clear
solution, upper thin black layer
5 Olive green upper layer and clear light blue lower layer
6Clear brown upper layer and very light clear blue lower
layer
7Green colored solution upper layer with clear blue lower
layer
8 No blue color formed at lower layer
9 Light blue solution at lower layer
10 Light blue colored solution at lower layer
11 Presence of the blue color in the lower layer
Group Creatinine Test
1 Clear orange colored solution formed
2 Clear red orange solution
3 Dark orange colored solution appear
4 Red orange colored solution formed
5 Clear red orange solution formed
6 Clear red orange solution formed
7 Clear red orange solution formed
8 Orange solution formed
9 Orange solution formed
10 Red colored solution
11 Red orange colored solution
Group Urine Sample Positive Control
1Black precipitate settled at
lower layer and cloudy layers visible on upper layer
Black precipitate was formed but disappeared. Turbid
solution with cloudy precipitates at bottom
2Dark yellow orange solution with light yellow precipitate
Dark yellow solution with light yellow precipitate
3Light yellow turbid solution
with black cloud formation seen at the lower layer/bottom
Pale turbid yellowish solution with light yellow precipitate
at the bottom; no black cloud solution appeared
4Clear light yellow solution with
cloudy precipitate Clear yellow solution with
yellow precipitate
5Formation of cloudy white
yellow solution
Formation of white cloudy precipitate with light yellow
orange solution
C. Examination for Pathological Constituents of Urine SampleC1. Gunning’s Test
Group Urine Sample Positive Control
6Cloudy yellow solution with
yellow suspension
Clear yellow orange solution with cloudy white suspension
at the lower part
7Turbid/cloudy solution with
precipitate on middle
Slightly turbid/cloudy yellow solution with black precipitate
at bottom
8
Clear red orange solution with cloudy white lower layer and red orange precipitate at the
bottom
Clear orange solution with cloudy white lower layer and yellow orange precipitate at
bottom
9Presence of black crystals
settled on the bottomPresence of black crystals
settled on the bottom
10Formation of black iodoform
crystals Turbid solution
11Yellow solution with cloudy red
precipitate on the bottomYellow solution with white precipitate on the bottom
C2. Benedict’s Test
Group Urine Sample Positive Control
1 Blue SolutionYellow-green turbid solution
with yellow precipitate settled on the bottom
2Semi turbid solution with green
precipitateTurbid orange solution with
orange precipitate
3
Blue-green colored solution appeared with formation of a cloudy-like substance at the lower part of the test tube
Two layers appeared: a dark yellow turbid upper layer solution and a greenish
colored solution for the lower layer; yellow powdered-like
precipitate formed
4 Clear blue solutionUpper yellow layer, middle
green layer with orange precipitate
5Pale orange precipitate with
blue-green interphase; yellow solution
Formation of a clear aqua solution with suspended
particles
Group Urine Sample Positive Control
6
Clear green upper layer and clear light blue lower layer with
some particles suspended in the lower layer and between 2
layers
Orange solution with green and orange precipitate
7
Orange solution upper layer; Green turbid solution middle
layer; Brownish orange precipitate at lower layer
Slightly clear blue solution with little precipitate
8Blue green turbid upper layer
with few scattered and settling white precipitate
Turbid yellowish solution through-out 4 layers with the third layer being blue green;
Greenish and yellow precipitate at the bottom
9 Clear blue solution Presence of brick red precipitate
10 Light blue solution Orange precipitate formed in an orange solution
11Blue solution with a little white
precipitate
Green turbid solution on top with orange precipitate on
the bottom
C3. Exton’s TestGroup Urine Sample Positive Control
1 Clear light yellow solution Creamy white solution with white precipitate
2
Turbid light orange solution with fibrous materials in upper layer and
turbid green with precipitate at lower later
Clear orange solution
3 Clear light yellow solutionCloudy pale yellow solution and
formation of light yellow precipitate at the upper surface
4 Clear solution Formation of cloudy mass suspended in a solution
5Formation of clear light yellow
solutionFormation of cloudy solution with
clear bottom
6 Clear light yellow-orange solution Light yellow solution with light yellow aggregates
7 Clear light yellow solution White turbid solution with white precipitate
8 Clear golden yellow solution White turbid solution
9 Clear solution Cloudiness is seen with mass of white precipitate
10 Clear colorless solution White cloudy present
11 No cloudiness appeared Cloudiness appeared
C4. Smith’s Test
Group Urine Sample Positive Control
1 Green interphase Green interphase
2 No emerald green interphase Emerald green interphase
3 Emerald green interphase Emerald green interphase
4 Yellow interphase Yellow interphase
5 No emerald green interphase No emerald green interphase
6 No emerald green interphase No emerald green interphase
7 No emerald green interphase No emerald green interphase
8 No emerald green interphase No emerald green interpahse
9No emerald green interphase
seen at point of contactNo emerald green interphase at
point of contact
10 No emerald green interphase Emerald green color appeared at point of contact
11Emerald green color is observed
at the point of contact
Emerald green color is observed at the point of
contact
C5. Test for Occult Blood
Group Urine Sample Positive Control
1 Blue green ring absent Blue green ring formed
2 Blue green ring absent Blue green ring formed
3 Blue green ring absent Dark blue colored solution appeared
4 Blue green ring absent Blue green ring formed
5 Blue green ring absent Blue green ring formed
6 Blue green ring absent Blue green ring formed
7 Blue green ring absent Blue green ring formed
8 Blue green ring absent Blue green ring formed
9 Blue green ring absent Blue green ring formed
10 Blue green ring absent Blue green ring formed
11 Blue green ring absent Blue green ring formed
DiscussionA. INITIAL EXAMINATION OF URINE SAMPLE
A. Initial Examination of Urine Sample
A1. Time of collection First voided morning specimen (most common)
Most ideal to test for substances Most concentrated Formed elements are more stable
Random urine sample (emergency) Collected at any time, usually daytime Exercise and excessive fluid intake can directly affect urine
composition
Clean-catch, midstream specimen (urine culture) Prior hydration of patient will lead to a “clean catch” urine
specimen
Time collected
Color Turbidity pH
19/20/12 7:15
amDark yellow Turbid 6
2 9/20/12 7:30 am
Yellow Orange Clear 5
3 9/20/12 9:00 am
Light Pale Yellow
Slightly Turbid 8
4 9/19/12 10:59 pm
Light Yellow Slightly turbid w/ minute suspended particles
6
5 9/20/12 6:00 am
Dark Yellow Clear 6
6 9/19/12 8:25 pm
Dark Yellow Clear 6
7 9/20/12 7:45 am
Light Yellow Clear 6
8 9/20/12 10:00 am
Yellow Orange Turbid 6
9 9/20/12 9:55 am
Very light yellow
Clear 6
10 9/20/12 9:20 am
Light Yellow Clear 7
11 9/20/12 9:30 am
Golden Yellow Clear 6
A. Initial Examination of Urine Sample
A2. Color Normal urine is clear and pale yellow Yellow color is due to the pigment urochrome
present Urine darkens on standing because of oxidation of
urobilinogen to urobilin Color variations indicate presence of metabolic
abnormality, diseases, stress, ingested foods or drug Colorless – dilution/too much hydration; Diabetes mellitus;
intake of drugs with diuretic effect Orange – excessive sweating, concentrated urine
Dark brown, brown red or dark yellow – very concentrated urine, acute febrile disease
Time collected
Color Turbidity pH
19/20/12 7:15
amDark yellow Turbid 6
2 9/20/12 7:30 am
Yellow Orange Clear 5
3 9/20/12 9:00 am
Light Pale Yellow
Slightly Turbid 8
4 9/19/12 10:59 pm
Light Yellow Slightly turbid w/ minute suspended particles
6
5 9/20/12 6:00 am
Dark Yellow Clear 6
6 9/19/12 8:25 pm
Dark Yellow Clear 6
7 9/20/12 7:45 am
Light Yellow Clear 6
8 9/20/12 10:00 am
Yellow Orange Turbid 6
9 9/20/12 9:55 am
Very light yellow
Clear 6
10 9/20/12 9:20 am
Light Yellow Clear 7
11 9/20/12 9:30 am
Golden Yellow Clear 6
A. Initial Examination of Urine Sample
A3. Turbidity
Urine may become cloudy due to the presence of amorphous phosphates which will disappear or due to urates in urine
Pathologic (e.g. Blood cells, fats, bacteria) – metabolic dysfunction, disease process, or deterioration of the barrier separating the urinary tract from the blood
Non-pathologic – examples are normal crystals, mucus, epithelial cells, spermatozoa and prostatic fluid , cellular casts
Time collected
Color Turbidity pH
19/20/12 7:15
amDark yellow Turbid 6
2 9/20/12 7:30 am
Yellow Orange Clear 5
3 9/20/12 9:00 am
Light Pale Yellow
Slightly Turbid 8
4 9/19/12 10:59 pm
Light Yellow Slightly turbid w/ minute suspended particles
6
5 9/20/12 6:00 am
Dark Yellow Clear 6
6 9/19/12 8:25 pm
Dark Yellow Clear 6
7 9/20/12 7:45 am
Light Yellow Clear 6
8 9/20/12 10:00 am
Yellow Orange Turbid 6
9 9/20/12 9:55 am
Very light yellow
Clear 6
10 9/20/12 9:20 am
Light Yellow Clear 7
11 9/20/12 9:30 am
Golden Yellow Clear 6
A. Gross physicochemical examination of urine sample
A4. Urine pH Normal urine pH varies from 4.5 – 8.0 Urinary pH becomes alkaline because CO2 will diffuse
into the air The kidney plays a major role in the regulation of
acid-base balance Reabsorption of sodium and tubular secretion of hydrogen
and ammonium ions Acidic – sodium concentration and excess acid retain by
body Alkaline – bicarbonate-carbonic acid buffer; normally
secreted when there is an excess of base or alkali in the body
Time collected
Color Turbidity pH
19/20/12 7:15
amDark yellow Turbid 6
2 9/20/12 7:30 am
Yellow Orange Clear 5
3 9/20/12 9:00 am
Light Pale Yellow
Slightly Turbid 8
4 9/19/12 10:59 pm
Light Yellow Slightly turbid w/ minute suspended particles
6
5 9/20/12 6:00 am
Dark Yellow Clear 6
6 9/19/12 8:25 pm
Dark Yellow Clear 6
7 9/20/12 7:45 am
Light Yellow Clear 6
8 9/20/12 10:00 am
Yellow Orange Turbid 6
9 9/20/12 9:55 am
Very light yellow
Clear 6
10 9/20/12 9:20 am
Light Yellow Clear 7
11 9/20/12 9:30 am
Golden Yellow Clear 6
B. QUALITATIVE EXAMINATION FOR NORMAL ORGANIC CONSTITUENTS OF URINE
DISCUSSION
TEST FOR UREAUrea The end product of protein catabolism Water-soluble compound Synthesized exclusively in the liver Via urea cycle:
involves conversion of ammonia into urea reactions occur in the liver, but urea is
transported to the kidneys where it is excreted
TEST FOR UREA
TEST FOR UREA Reagents
70% NaOH, bromine water
Principle Hydrolyzation of urea with NaOH Excess NaOH then reacts with bromine water to
form NaOBr, which in turn oxidizes the urea to form N2,CO2, H2O
Positive result Evolution of N2 gas as indicated with presence of
bubbles
TEST FOR UREA
MECHANISM
Group Test for Urea
1White gas was observed then brown gas almost
immediately
2Clear yellow solution upper layer and clear light yellow
lower layer
3Light yellow solution, with N2
gases, presence of cloudy white substance
4 Formation of bubbles, light yellow solution
5 Formation of bubbles on top
6Clear yellow with formation of bubbles on the
uppermost portion
7Clear yellow solution with presence of bubbles and
foamy layer
8Light yellow color upper layer with white bubbles and
clear yellow second layer
9Yellow solution with presence of bubbles indicating
formation of N2
gas
10 Evolution of gas was observed
11There was an evolution of N2
gas
TEST FOR UREA
TEST FOR UREA
INDICATIONS Levels of urea depend upon protein
intake, protein catabolism and kidney function
Amount of urea excreted per day is 30 g by a normal adult
Elevated urea levels can occur because of: Diseases which impair kidney function,
congestive heart failure, diabetes, dietary changes, liver diseases
TEST FOR URIC ACID
Uric acid Major end product of catabolism of
purine bases – adenine and guanine nucleotides of cellular DNA and RNA
Formed from dietary nucleic acids Dissolves in blood and travels to the
kidneys, where it passes out in urine Excreted in only small amounts in urine Formula: C5H4N4O3
TEST FOR URIC ACID Reagents used
20%Na2CO3, phosphotungstic acid reagent
Principle Oxidation of alkene in uric acid by
phosphotungstic acid(protein precipitant) reagent, yielding allantoin and tungsten blue
Positive result Blue solution
TEST FOR URIC ACID
Reaction Mechanism for the Test for Uric Acid
Group Test for Uric Acid
1 Clear blue solution
2 Clear light blue solution
3 A light blue colored solution appeared
4 Clear blue solution
5 Formation of blue clear solution
6 Clear blue solution
7 Clear light blue solution
8 Blue solution formed
9 Light blue solution
10 Light blue colored solution
11 Clear blue solution
TEST FOR URIC ACID
TEST FOR URIC ACIDINDICATIONS High levels of uric acid can cause
kidney stones Uric acid excretion is 250-750 mg per
day Level of uric acid gradually increases
with age High levels may be due to:
A high purine diet, gout, and cortisone therapy. Gout – a painful condition in which high
blood levels of uric acid in the body can cause solid crystals to form within joints
INDICAN TEST
Indican An indole produced by bacterial action
on amino acid, tryptophan, in the intestine
Reflects bacterial activity in the intestines
Detection in urine depends upon its decomposition and subsequent oxidation of indoxyl to indigo blue
Most is excreted in the feces
INDICAN TEST Reagent
Obermayer’s reagent (FeCl3 in concentrated HCl and chloroform)
Principle Oxidation of indoxyl to blue indigo and its
absorption into the chloroform layer Positive result
Blue color of the chloroform layer
INDICAN TESTMECHANISM
Group Indican Test
1Clear blue colored lower layer and dark green upper
layer
2Clear brownish green upper layer and blue lower layer
with presence of emulsion
3Olive green solution at upper layer and pale clear blue
solution at lower layer
4Lower light blue clear solution, middle olive green
clear solution, upper thin black layer
5 Olive green upper layer and clear light blue lower layer
6Clear brown upper layer and very light clear blue lower
layer
7Green colored solution upper layer with clear blue lower
layer
8 No blue color formed at lower layer
9 Light blue solution at lower layer
10 Light blue colored solution at lower layer
11 Presence of the blue color in the lower layer
INDICAN TEST
INDICAN TESTINDICATIONS
Increased with high protein diets or inefficient protein digestion
Excessive urine indicant include: Maldigestion or malabsorption of
Tryptophan or increased bacterial overgrowth which reflects enteric toxemia
CREATININE TESTCreatinine Anhydride of creatine A constant constituent of normal urine Formed from creatine phosphate which is
concerned with muscle contraction Breakdown of phospocreatine by the addition
of strong acid or alkali. Creatine phosphate loses phosphoric acid and creatine undergoes dehydration forming creatinine
CREATININE TEST Reagents:
Alkali picrate solution (5:1, saturated picric acid and 10% NaOH).
Principle: Creatinine reacts with picric acid to form
creatinine picrate complex. Positive result:
Red-orange colored solution.
CREATININE TESTMECHANISM
Group Creatinine Test
1 Clear orange colored solution formed
2 Clear red orange solution
3 Dark orange colored solution appear
4 Red orange colored solution formed
5 Clear red orange solution formed
6 Clear red orange solution formed
7 Clear red orange solution formed
8 Orange solution formed
9 Orange solution formed
10 Red colored solution
11 Red orange colored solution
CREATININE TEST
INDICATIONS All are positive results because creatinine picrate is orange in
color. If the result was red-orange solution, there is a high presence of creatinine excreted in the urine. However, if the color is lighter, it means that there is less creatinine in urine and more in blood because it was not filtered properly.
Decrease of creatinine levels may be due to: Muscular dystrophy
Increase of creatinine levels may be due to: Dehydration, bacterial infection, shock, kidney failure,
kidney stones, atherosclerosis
CREATININE TEST
C. QUALITATIVE EXAMINATION FOR PATHOLOGICAL ORGANIC CONSTITUENTS OF URINE
DISCUSSION
GUNNING’S TESTKETONE BODIES
Are products of incomplete fat metabolism Serve as energy source used in the citric
cycle in place of glucose (when insufficient) Three water soluble compounds that are
produced as by products when fatty acids are broken down for energy in the body
GUNNING’S TEST Test for Ketone bodies Reagents Used:
Acetone (for positive control only), Conc. Ammonium hydroxide, Lugol’s solution
Principle Halogenation of the enolate of acetone
enolate formation is catalyzed by presence of hydroxyl ions from NH4OH
Positive result Iodoform crystals
GUNNING’S TESTMECHANISM
Group Urine Sample Positive Control
1Black precipitate settled at lower layer and cloudy layers
visible on upper layer
Black precipitate was formed but disappeared.
Turbid solution with cloudy precipitates at bottom
2Dark yellow orange solution
with light yellow precipitateDark yellow solution with light yellow precipitate
3Light yellow turbid solution
with black cloud formation seen at the lower layer/bottom
Pale turbid yellowish solution with light yellow
precipitate at the bottom; no black cloud solution
appeared
4Clear light yellow solution with
cloudy precipitate Clear yellow solution with
yellow precipitate
5Formation of cloudy white
yellow solution
Formation of white cloudy precipitate with light yellow
orange solution
GUNNING’S TEST
Group Urine Sample Positive Control
6Cloudy yellow solution with
yellow suspension
Clear yellow orange solution with cloudy white
suspension at the lower part
7Turbid/cloudy solution with
precipitate on middle
Slightly turbid/cloudy yellow solution with black
precipitate at bottom
8
Clear red orange solution with cloudy white lower layer and
red orange precipitate at the bottom
Clear orange solution with cloudy white lower layer and yellow orange precipitate
at bottom
9Presence of black crystals
settled on the bottomPresence of black crystals
settled on the bottom
10Formation of black iodoform
crystals Turbid solution
11Yellow solution with cloudy red
precipitate on the bottomYellow solution with white precipitate on the bottom
GUNNING’S TEST
GUNNING’S TESTINDICATIONS
Ketonuria – ketone bodies present in urine
Ketosis – high level of ketone in blood, distinct odor
Ketogenesis – when liver glycogen is depleted
Ketoacidosis – insulin deficiency, Type 1 diabetes
BENEDICT’S TEST
GLUCOSE a simple monosaccharide found in plants serve as an energy source Usually, there is little or no presence of
glucose in the urine Glycosuria
Presence of glucose in the urine
BENEDICT’S TEST (TEST FOR GLUCOSE)
Reagents Benedict’s reagent (carbonate-citrate solution and
copper sulfate solution)
Principle Oxidation reaction of glucose by the Cu2
+ and reduction of Cu2+ to Cu+
Positive Result Formation of brick red precipitate
BENEDICT’S TESTMECHANISM
C2. Benedict’s Test
Group Urine Sample Positive Control
1 Blue SolutionYellow-green turbid solution
with yellow precipitate settled on the bottom
2Semi turbid solution with green
precipitateTurbid orange solution with
orange precipitate
3
Blue-green colored solution appeared with formation of a cloudy-like substance at the lower part of the test tube
Two layers appeared: a dark yellow turbid upper layer solution and a greenish
colored solution for the lower layer; yellow powdered-like
precipitate formed
4 Clear blue solutionUpper yellow layer, middle
green layer with orange precipitate
5Pale orange precipitate with
blue-green interphase; yellow solution
Formation of a clear aqua solution with suspended
particles
Group Urine Sample Positive Control
6
Clear green upper layer and clear light blue lower layer with
some particles suspended in the lower layer and between 2
layers
Orange solution with green and orange precipitate
7
Orange solution upper layer; Green turbid solution middle
layer; Brownish orange precipitate at lower layer
Slightly clear blue solution with little precipitate
8Blue green turbid upper layer
with few scattered and settling white precipitate
Turbid yellowish solution through-out 4 layers with the third layer being blue green;
Greenish and yellow precipitate at the bottom
9 Clear blue solution Presence of brick red precipitate
10 Light blue solution Orange precipitate formed in an orange solution
11Blue solution with a little white
precipitate
Green turbid solution on top with orange precipitate on
the bottom
BENEDICT’S TESTINDICATIONS:
180mg/dL glucose in urine is the normal glucose level of a person. 300-500 mg/dL are common with severe untreated diabetes
Positive result indicates that glucose is present in the urine. Presence of glucose can be due to: Diabetes mellitus; Renal glycosuria
EXTON’S TEST
ALBUMIN Is an umbrella term for a type of protein
which is water soluble protein is not usually found in urine Albumin in the human body transports
essential fatty acids from adipose tissue, otherwise known as fat, to muscle tissue
EXTON’S TEST Test for Albumin Reagents
Exton’s reagent (anhydrous sodium sulfate and sulfosalicylic acid(SSA) dissolved in distilled water)
Principle Na2SO4 and SSA will cause acidification and
the application of heat precipitates albumin
Positive Result Formation of a white turbid and cloudy
precipitate
Exton’s Test
Group Urine Sample Positive Control
1 Clear light yellow solution Creamy white solution with white precipitate
2Turbid light orange solution with fibrous
materials in upper layer and turbid green with precipitate at lower later
Clear orange solution
3 Clear light yellow solutionCloudy pale yellow solution and
formation of light yellow precipitate at the upper surface
4 Clear solution Formation of cloudy mass suspended in a solution
5Formation of clear light yellow
solutionFormation of cloudy solution with
clear bottom
6 Clear light yellow-orange solution Light yellow solution with light yellow aggregates
7 Clear light yellow solution White turbid solution with white precipitate
8 Clear golden yellow solution White turbid solution
9 Clear solution Cloudiness is seen with mass of white precipitate
10 Clear colorless solution White cloudy present
11 No cloudiness appeared Cloudiness appeared
EXTON’S TESTINDICATIONS
Albuminuria Presence of albumin in the urine
Proteins from the blood can leak into the urine when the glomerular filtration unit of the kidney are damaged/severed
SMITH’S TESTBILE PIGMENTS
responsible for lipid catabolism
BILIRUBIN It is a yellow pigment found in bile, a fluid made
by the liver. It is secreted in bile and urine. It is responsible for the brown coloration of feces
and yellow coloration of urine, bruises and jaundice.
Major constituent of bile. Conjugated bilirubin is water-soluble and can
therefore be excreted in urine. Unconjugated bilirubin is water-insoluble.
SMITH’S TEST Test for Bile Pigments Reagents
Tincture of alcoholic iodine mixture (iodine crystals and sodium iodide in an absolute ethanol, then added with distilled water)
Principle Oxidation of the bile pigments to colored
derivatives of the alcoholic iodide Positive Result
Formation of emerald green at the point of contact
Smith’s Test
Group Urine Sample Positive Control
1 Green interphase Green interphase
2 No emerald green interphase Emerald green interphase
3 Emerald green interphase Emerald green interphase
4 Yellow interphase Yellow interphase
5 No emerald green interphase No emerald green interphase
6 No emerald green interphase No emerald green interphase
7 No emerald green interphase No emerald green interphase
8 No emerald green interphase No emerald green interphase
9No emerald green interphase
seen at point of contactNo emerald green interphase at
point of contact
10 No emerald green interphase Emerald green color appeared at point of contact
11Emerald green color is observed
at the point of contact
Emerald green color is observed at the point of
contact
SMITH’S TESTINDICATIONS Dark urine is caused when there is obstruction
to bile flow, or obstructive jaundice. In which case there is direct bilirubinemia, which is water soluble and is filtered through the glomerular membrane. This is the cause of dark urine.
High levels of bile pigment (hyperbilirubinemia) in urine may be due to: Liver injury Presence of gallstones Drug toxicity Hemolytic anemia
TEST FOR OCCULT BLOODOCCULT BLOOD sometimes referred to as “hidden blood”
Heme groups are most commonly recognized in their presence as components of hemoglobin (red pigment in blood)
Can be either hemolyzed (dissolved blood) or non-hemolyzed (intact red blood cells)
TEST FOR OCCULT BLOOD Reagents
½ spatula of guaiac poweder in 5 mL 95% ethanol solution; hydrogen peroxide
Principle The peroxidase activity of the Heme which
decomposes H2O2 and the liberated oxygen oxidizes organic substances such as guaiac powder
Positive Result Presence of orange, green ring Blue ring if much or excess blood is detected
TEST FOR OCCULT BLOODMECHANISM
Test for Occult Blood
Group Urine Sample Positive Control
1 Blue ring absent Blue green ring formed
2Blue ring absent
Blue green ring formed
3Blue ring absent
Dark blue colored solution appeared
4Blue ring absent
Blue green ring formed
5Blue ring absent
Blue green ring formed
6Blue ring absent
Blue green ring formed
7Blue ring absent
Blue green ring formed
8Blue ring absent
Blue green ring formed
9Blue ring absent
Blue green ring formed
10Blue ring absent
Blue green ring formed
11Blue ring absent
Blue green ring formed
TEST FOR OCCULT BLOODINDICATIONS Hematuria
The presence of more or less intact red blood cells in the urine
Usually accompanied by hemoglobinuria because of disintegration of red cell in urine or brown or produce a smoky appearance
CONCLUSION Urine samples from the 11 groups obtained a pH
range of 5-8, most are not turbid and samples in which suspended particles were seen are collected for almost 12 hours prior to the examination that could otherwise allowed the particles to settled down and provides a false observation and color of samples varies from very light yellow to dark orange with respect to concentration.
Qualitative examination for normal organic constituents of urine proved that normally it contains Urea, Uric Acid, Indican and Creatinine.
CONCLUSION
Most urine were found to be negative for presence of albumin, occult blood, ketone bodies and bile pigments(with minor exception for sample nos. 1, 3 and 11.). However 7 out of 11 samples were proven to be positive for presence of glucose indicating high sugar consumption or diet.