Urinalysis

Jorge Anton D. Ordas Eloisah Vin S. Ragodon Lindley C. Susi Mercer Keith Von Possel Mark 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. 2. 3.

Prerenal dehydration, congestive heart failure and hemorrhage Postrenal obstruction of the urinary tract 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
-add

0.5 mL 70% NaOH and 4 drops bromine water

Observe formation of bubbles

Procedure/Methodology
B2. Test for uric acid
1 mL Urine Sample

-add 5 mL of 20% Na2CO3 , then mix -add 5 drops phosphotungstic acid reagent, mix -observethen for formation of blue solution
Record Result

Procedure/Methodology
B.3 Indican test
5 mL Urine Sample

5 mL Urine Sample

- add 5 mL Obermayers Reagent, then mix well - add 3 mL chloroform. Shake and allow chloroform to settle. for formation of - observe blue color in lower layer
RecordResult Result Record

Procedure
B4. Creatinine test
2 mL Urine Sample

-add 1 mL of alkaline picrate solution

-note for formation of an orange solution

Record Result

Procedure
C. Examination of pathologic organic constituents Controltest C1. Positive Gunnings
5 mL Urine + 2 mL acetone 5 mL Urine Sample

-Basify with 5 drops concentrated ammonium hydroxide .Check pH by red litmus paper. - add Lugols solution to sample until black cloud appears - note the formation of iodoform crystals
Record Result

Procedure
C.2. Benedicts test
Positive Control

1 mL Urine + 1 mL glucose solution

0.5 mL Urine Sample

- add 5 mL of Benedicts reagent - heat in boiling water bath for 2-3 minutes then cool - note precipitate formation
Record Result

Procedure
C3. Extons test
Positive Control

3 mL Urine + pinch of albumin

3 mL Urine Sample

-add 3 mL of Extons reagent

-warm if cloudiness appears -if cloudiness persist or increase in heating, albumin is present
Record Result

Procedure
C4. Smiths test
Positive Control

3 mL Urine + 3-5 drops bile pigment

5 mL Urine Sample

Note if emerald green is seen at point of contact

-incline test tube and overlay with 3 mL of tincture of alcoholic iodine mixture

Procedure
C5. Test for occult blood
Positive Control

3 mL acidified Urine + 3 drops of blood

3 mL Acidified Urine Sample

- add 5 mL of 95% ethanol to half spatula guaiac powder then add 5 mL of hydrogen peroxide in one test tube - add 5 mL of this solution to 3 mL acidified urine and positive conrol - note the formation of iodoform crystals
Record Result

Results

A. Initial Examination of Urine Sample

Time collected 1 2 3 4 9/20/12 7:15 am 9/20/12 7:30 am 9/20/12 9:00 am 9/19/12 10:59 pm Color Dark yellow Yellow Orange Light Pale Yellow Light Yellow Turbidity Turbid Clear Slightly Turbid Slightly turbid w/ minute suspended particles Clear Clear Clear Turbid Clear Clear Clear pH 6 5 8 6

5 6 7 8 9 10 11

9/20/12 6:00 am 9/19/12 8:25 pm 9/20/12 7:45 am 9/20/12 10:00 am 9/20/12 9:55 am 9/20/12 9:20 am 9/20/12 9:30 am

Dark Yellow Dark Yellow Light Yellow Yellow Orange Very light yellow Light Yellow Golden Yellow

6 6 6 6 6 7 6

B. Examination for Normal Organic Constituents of Urine Sample Group Test for Urea
1 2 3 4 5

White gas was observed then brown gas almost immediately

Clear yellow solution upper layer and clear light yellow lower layer
Light yellow solution, with N2 gases, presence of cloudy white substance Formation of bubbles, light yellow solution Formation of bubbles on top

6
7 8 9 10 11

Clear yellow solution with formation of bubbles on the uppermost portion

Clear yellow solution with presence of bubbles and foamy layer Light yellow color upper layer with white fumes and clear yellow second layer Yellow solution with presence of bubbles indicating formation of N2 gas Evolution of gas was observed There was an evolution of N2 gas

Group 1 2 3 4 5 6 7 8 9 10 11

Test for Uric Acid

Clear blue solution Clear light blue solution light blue colored solution appeared Clear blue solution Formation of blue clear solution Clear blue solution Clear light blue solution Blue solution formed Light blue solution Light blue colored solution Clear blue solution

Group 1 2 3 4 5 6 7 8 9 10 11

Indican Test

Clear blue colored lower layer and dark green upper layer Clear brownish green upper layer and blue lower layer with presence of emulsion Olive green solution at upper layer and pale clear blue solution at lower layer Lower light blue clear solution, middle olive green clear solution, upper thin black layer Olive green upper layer and clear light blue lower layer Clear brown upper layer and very light clear blue lower layer Green colored solution upper layer with clear blue lower layer No blue color formed at lower layer Light blue solution at lower layer Light blue colored solution at lower layer Presence of the blue color in the lower layer

Group 1 2 3 4 5 6 7 8 9 10 11

Creatinine Test

Clear orange colored solution formed Clear red orange solution Dark orange colored solution appear

Red orange colored solution formed

Clear red orange solution formed Clear red orange solution formed Clear red orange solution formed Orange solution formed Orange solution formed Red colored solution Red orange colored solution

C. Examination for Pathological Constituents of Urine Sample C1. Gunnings Test

Group Urine Sample Positive Control

Black precipitate settled at lower layer and cloudy layers visible on upper layer Dark yellow orange solution with light yellow precipitate

Black precipitate was formed but disappeared. Turbid solution with cloudy precipitates at bottom Dark yellow solution with light yellow precipitate Pale turbid yellowish solution with light yellow precipitate at the bottom; no black cloud solution appeared Clear yellow solution with yellow precipitate Formation of white cloudy precipitate with light yellow orange solution

Light yellow turbid solution with black cloud formation seen at the lower layer/bottom
Clear light yellow solution with cloudy precipitate Formation of cloudy white yellow solution

Group

Urine Sample

Positive Control

Cloudy yellow solution with yellow suspension Turbid/cloudy solution with precipitate on middle Clear red orange solution with cloudy white lower layer and red orange precipitate at the bottom Presence of black crystals settled on the bottom Formation of black iodoform crystals Yellow solution with cloudy red precipitate on the bottom

Clear yellow orange solution with cloudy white suspension at the lower part Slightly turbid/cloudy yellow solution with black precipitate at bottom Clear orange solution with cloudy white lower layer and yellow orange precipitate at bottom Presence of black crystals settled on the bottom

10

Turbid solution
Yellow solution with white precipitate on the bottom

11

C2. Benedicts Test

Group Urine Sample Positive Control

Blue Solution Semi turbid solution with green precipitate Blue-green colored solution appeared with formation of a cloudy-like substance at the lower part of the test tube

Yellow-green turbid solution with yellow precipitate settled on the bottom Turbid orange solution with orange precipitate Two layers appeared: a dark yellow turbid upper layer solution and a greenish colored solution for the lower layer; yellow powdered-like precipitate formed Upper yellow layer, middle green layer with orange precipitate Formation of a clear aqua solution with suspended particles

Clear blue solution

Pale orange precipitate with bluegreen interphase; yellow solution

Group

Urine Sample

Positive Control

Clear green upper layer and clear light blue lower layer with some particles suspended in the lower layer and between 2 layers Orange solution upper layer; Green turbid solution middle layer; Brownish orange precipitate at lower layer Blue green turbid upper layer with few scattered and settling white precipitate Clear blue solution Light blue solution Blue solution with a little white precipitate

Orange solution with green and orange precipitate Slightly clear blue solution with little precipitate Turbid yellowish solution throughout 4 layers with the third layer being blue green; Greenish and yellow precipitate at the bottom Presence of brick red precipitate Orange precipitate formed in an orange solution Green turbid solution on top with orange precipitate on the bottom

9 10

11

C3. Extons Test

Group 1 Urine Sample Positive Control

Clear light yellow solution

Turbid light orange solution with fibrous materials in upper layer and turbid green with precipitate at lower later

Creamy white solution with white precipitate Clear orange solution

Clear light yellow solution

Cloudy pale yellow solution and formation of light yellow precipitate at the upper surface
Formation of cloudy mass suspended in a solution

4 5 6 7 8 9 10 11

Clear solution Formation of clear light yellow solution Clear light yellow-orange solution Clear light yellow solution Clear golden yellow solution Clear solution Clear colorless solution

Formation of cloudy solution with clear bottom

Light yellow solution with light yellow aggregates White turbid solution with white precipitate White turbid solution Cloudiness is seen with mass of white precipitate White cloudy present

No cloudiness appeared

Cloudiness appeared

C4. Smiths Test

Group 1 2 3 4 5 6 7 8 9 Urine Sample Positive Control

Green interphase

Green interphase

No emerald green interphase

Emerald green interphase Yellow interphase No emerald green interphase No emerald green interphase No emerald green interphase No emerald green interphase No emerald green interphase seen at point of contact No emerald green interphase Emerald green color is observed at the point of contact

Emerald green interphase

Emerald green interphase Yellow interphase No emerald green interphase No emerald green interphase No emerald green interphase No emerald green interpahse No emerald green interphase at point of contact Emerald green color appeared at point of contact Emerald green color is observed at the point of contact

10

11

C5. Test for Occult Blood

Group 1 2 3 4 5 6 7 8 9 10 11

Urine Sample

Positive Control

Blue green ring absent

Blue green ring formed

Blue green ring absent

Blue green ring absent Blue green ring absent Blue green ring absent Blue green ring absent Blue green ring absent Blue green ring absent Blue green ring absent Blue green ring absent Blue green ring absent

Blue green ring formed

Dark blue colored solution appeared Blue green ring formed Blue green ring formed Blue green ring formed Blue green ring formed Blue green ring formed Blue green ring formed Blue green ring formed Blue green ring formed

Discussion
A. 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 1 2 9/20/12 7:15 am 9/20/12 7:30 am

Color Dark yellow Yellow Orange

Turbidity Turbid Clear

pH 6 5

3
4

9/20/12 9:00 am
9/19/12 10:59 pm

Light Pale Yellow

Light Yellow

Slightly Turbid
Slightly turbid w/ minute suspended particles Clear Clear Clear Turbid Clear Clear Clear

8
6

5 6 7 8 9 10 11

9/20/12 6:00 am 9/19/12 8:25 pm 9/20/12 7:45 am 9/20/12 10:00 am 9/20/12 9:55 am 9/20/12 9:20 am 9/20/12 9:30 am

Dark Yellow Dark Yellow Light Yellow Yellow Orange Very light yellow Light Yellow Golden Yellow

6 6 6 6 6 7 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 1 2 3 4 9/20/12 7:15 am 9/20/12 7:30 am 9/20/12 9:00 am 9/19/12 10:59 pm

Color Dark yellow Yellow Orange Light Pale Yellow Light Yellow

Turbidity Turbid Clear Slightly Turbid Slightly turbid w/ minute suspended particles Clear Clear Clear Turbid Clear Clear Clear

pH 6 5 8 6

5 6 7 8 9 10 11

9/20/12 6:00 am 9/19/12 8:25 pm 9/20/12 7:45 am 9/20/12 10:00 am 9/20/12 9:55 am 9/20/12 9:20 am 9/20/12 9:30 am

Dark Yellow Dark Yellow Light Yellow Yellow Orange Very light yellow Light Yellow Golden Yellow

6 6 6 6 6 7 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 1 2 9/20/12 7:15 am 9/20/12 7:30 am

Color Dark yellow Yellow Orange

Turbidity Turbid Clear

pH 6 5

3
4

9/20/12 9:00 am
9/19/12 10:59 pm

Light Pale Yellow

Light Yellow

Slightly Turbid
Slightly turbid w/ minute suspended particles Clear Clear Clear Turbid Clear Clear Clear

8
6

5 6 7 8 9 10 11

9/20/12 6:00 am 9/19/12 8:25 pm 9/20/12 7:45 am 9/20/12 10:00 am 9/20/12 9:55 am 9/20/12 9:20 am 9/20/12 9:30 am

Dark Yellow Dark Yellow Light Yellow Yellow Orange Very light yellow Light Yellow Golden Yellow

6 6 6 6 6 7 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 1 2 3 4 9/20/12 7:15 am 9/20/12 7:30 am 9/20/12 9:00 am 9/19/12 10:59 pm

Color Dark yellow Yellow Orange Light Pale Yellow Light Yellow

Turbidity Turbid Clear Slightly Turbid Slightly turbid w/ minute suspended particles Clear Clear Clear Turbid Clear Clear Clear

pH 6 5 8 6

5 6 7 8 9 10 11

9/20/12 6:00 am 9/19/12 8:25 pm 9/20/12 7:45 am 9/20/12 10:00 am 9/20/12 9:55 am 9/20/12 9:20 am 9/20/12 9:30 am

Dark Yellow Dark Yellow Light Yellow Yellow Orange Very light yellow Light Yellow Golden Yellow

6 6 6 6 6 7 6

DISCUSSION
B. QUALITATIVE EXAMINATION FOR NORMAL ORGANIC CONSTITUENTS OF URINE

TEST FOR UREA

Urea

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 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

Principle

Positive result

Evolution of N2 gas as indicated with presence of bubbles

TEST FOR UREA

MECHANISM

TEST FOR UREA

Group 1 Test for Urea

White gas was observed then brown gas almost immediately

2
3 4 5

Clear yellow solution upper layer and clear light yellow lower layer
Light yellow solution, with N2 gases, presence of cloudy white substance Formation of bubbles, light yellow solution Formation of bubbles on top

6
7 8

Clear yellow with formation of bubbles on the uppermost portion

Clear yellow solution with presence of bubbles and foamy layer Light yellow color upper layer with white bubbles and clear yellow second layer

Yellow solution with presence of bubbles indicating formation of

9

N2
gas Evolution of gas was observed There was an evolution of N2 gas

10

11

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: C H N O
5 4 4 3

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

TEST FOR URIC ACID

Group 1 2 3 4 5 6 7 8 9 10 11 Test for Uric Acid

Clear blue solution Clear light blue solution A light blue colored solution appeared Clear blue solution Formation of blue clear solution Clear blue solution Clear light blue solution Blue solution formed Light blue solution Light blue colored solution Clear blue solution

TEST FOR URIC ACID

INDICATIONS 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

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
Obermayers

reagent (FeCl3 in concentrated HCl and chloroform)

Principle
Oxidation

of indoxyl to blue indigo and its absorption into the chloroform layer color of the chloroform layer

Positive result
Blue

INDICAN TEST
MECHANISM

INDICAN TEST
Group 1 2 3 4 5 6 7 8 9 10 11 Indican Test

Clear blue colored lower layer and dark green upper layer Clear brownish green upper layer and blue lower layer with presence of emulsion

Olive green solution at upper layer and pale clear blue solution at lower layer
Lower light blue clear solution, middle olive green clear solution, upper thin black layer Olive green upper layer and clear light blue lower layer Clear brown upper layer and very light clear blue lower layer Green colored solution upper layer with clear blue lower layer No blue color formed at lower layer Light blue solution at lower layer

Light blue colored solution at lower layer Presence of the blue color in the lower layer

INDICAN TEST
INDICATIONS

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 TEST
Creatinine

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. colored solution.

Positive result:
Red-orange

CREATININE TEST
MECHANISM

CREATININE TEST
Group 1 2 3 4 5 6 7 8 9 10 11 Creatinine Test

Clear orange colored solution formed Clear red orange solution Dark orange colored solution appear

Red orange colored solution formed

Clear red orange solution formed Clear red orange solution formed Clear red orange solution formed Orange solution formed Orange solution formed Red colored solution 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

DISCUSSION
C. QUALITATIVE EXAMINATION FOR PATHOLOGICAL ORGANIC CONSTITUENTS OF URINE

GUNNINGS TEST
KETONE 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

GUNNINGS TEST
Test for Ketone bodies Reagents Used:

Acetone

(for positive control only), Conc. Ammonium hydroxide, Lugols solution of the enolate of acetone

Principle
Halogenation
enolate

formation is catalyzed by presence of hydroxyl ions from NH4OH

Positive result

Iodoform crystals

GUNNINGS TEST
MECHANISM

GUNNINGS TEST
Group Urine Sample Positive Control

Black precipitate settled at lower layer and cloudy layers visible on upper layer Dark yellow orange solution with light yellow precipitate

Black precipitate was formed but disappeared. Turbid solution with cloudy precipitates at bottom Dark yellow solution with light yellow precipitate Pale turbid yellowish solution with light yellow precipitate at the bottom; no black cloud solution appeared

Light yellow turbid solution with black cloud formation seen at the lower layer/bottom Clear light yellow solution with cloudy precipitate
Formation of cloudy white yellow solution

Clear yellow solution with yellow precipitate

Formation of white cloudy precipitate with light yellow orange solution

GUNNINGS TEST
Group

Urine Sample

Positive Control

Cloudy yellow solution with yellow suspension

Clear yellow orange solution with cloudy white suspension at the lower part Slightly turbid/cloudy yellow solution with black precipitate at bottom
Clear orange solution with cloudy white lower layer and yellow orange precipitate at bottom Presence of black crystals settled on the bottom Turbid solution Yellow solution with white precipitate on the bottom

Turbid/cloudy solution with precipitate on middle

Clear red orange solution with cloudy white lower layer and red orange precipitate at the bottom Presence of black crystals settled on the bottom

10

Formation of black iodoform crystals

Yellow solution with cloudy red precipitate on the bottom

11

GUNNINGS TEST
INDICATIONS

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

BENEDICTS 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

BENEDICTS TEST (TEST FOR GLUCOSE)

Reagents

Benedicts reagent (carbonate-citrate solution and copper sulfate solution) Oxidation reaction of glucose by the Cu2+ and reduction of Cu2+ to Cu+ Formation of brick red precipitate

Principle

Positive Result

BENEDICTS TEST
MECHANISM

C2. Benedicts Test

Group Urine Sample Positive Control

Blue Solution Semi turbid solution with green precipitate Blue-green colored solution appeared with formation of a cloudy-like substance at the lower part of the test tube

Yellow-green turbid solution with yellow precipitate settled on the bottom Turbid orange solution with orange precipitate Two layers appeared: a dark yellow turbid upper layer solution and a greenish colored solution for the lower layer; yellow powdered-like precipitate formed Upper yellow layer, middle green layer with orange precipitate Formation of a clear aqua solution with suspended particles

Clear blue solution

Pale orange precipitate with bluegreen interphase; yellow solution

Group

Urine Sample

Positive Control

Clear green upper layer and clear light blue lower layer with some particles suspended in the lower layer and between 2 layers Orange solution upper layer; Green turbid solution middle layer; Brownish orange precipitate at lower layer

Orange solution with green and orange precipitate Slightly clear blue solution with little precipitate Turbid yellowish solution throughout 4 layers with the third layer being blue green; Greenish and yellow precipitate at the bottom Presence of brick red precipitate

Blue green turbid upper layer with few scattered and settling white precipitate
Clear blue solution Light blue solution Blue solution with a little white precipitate

9 10

Orange precipitate formed in an orange solution

Green turbid solution on top with orange precipitate on the bottom

11

BENEDICTS TEST
INDICATIONS:

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

EXTONS 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

EXTONS TEST
Test for Albumin Reagents

Extons

reagent (anhydrous sodium sulfate and sulfosalicylic acid(SSA) dissolved in distilled water) and SSA will cause acidification and the application of heat precipitates albumin of a white turbid and cloudy

Principle
Na2SO4

Positive Result
Formation

precipitate

Extons Test
Group 1 Urine Sample Positive Control

Clear light yellow solution

Turbid light orange solution with fibrous materials in upper layer and turbid green with precipitate at lower later

Creamy white solution with white precipitate Clear orange solution Cloudy pale yellow solution and formation of light yellow precipitate at the upper surface Formation of cloudy mass suspended in a solution

Clear light yellow solution

4 5 6 7 8 9 10 11

Clear solution Formation of clear light yellow solution Clear light yellow-orange solution Clear light yellow solution Clear golden yellow solution Clear solution Clear colorless solution No cloudiness appeared

Formation of cloudy solution with clear bottom

Light yellow solution with light yellow aggregates White turbid solution with white precipitate White turbid solution Cloudiness is seen with mass of white precipitate White cloudy present Cloudiness appeared

EXTONS TEST
INDICATIONS

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

SMITHS TEST
BILE 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.

SMITHS 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)
of the bile pigments to colored derivatives of the alcoholic iodide of emerald green at the point of

Principle
Oxidation

Positive Result
Formation

contact

Smiths Test
Group 1 2 3 4 5 6 7 8 9 Urine Sample Positive Control

Green interphase

Green interphase

No emerald green interphase

Emerald green interphase Yellow interphase No emerald green interphase No emerald green interphase No emerald green interphase No emerald green interphase No emerald green interphase seen at point of contact No emerald green interphase Emerald green color is observed at the point of contact

Emerald green interphase

Emerald green interphase Yellow interphase No emerald green interphase No emerald green interphase No emerald green interphase No emerald green interphase No emerald green interphase at point of contact Emerald green color appeared at point of contact Emerald green color is observed at the point of contact

10

11

SMITHS TEST
INDICATIONS

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 BLOOD

OCCULT 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 nonhemolyzed (intact red blood cells)

TEST FOR OCCULT BLOOD

Reagents

spatula of guaiac poweder in 5 mL 95% ethanol solution; hydrogen peroxide peroxidase activity of the Heme which decomposes H2O2 and the liberated oxygen oxidizes organic substances such as guaiac powder

Principle
The

Positive Result
Presence

of orange, green ring Blue ring if much or excess blood is detected

TEST FOR OCCULT BLOOD

MECHANISM

Test for Occult Blood

Group 1 2 3 4 5 Urine Sample Blue ring absent Blue ring absent Blue ring absent Blue ring absent Blue ring absent Blue ring absent Blue ring absent Blue ring absent Blue ring absent Blue ring absent Blue ring absent Positive Control Blue green ring formed Blue green ring formed Dark blue colored solution appeared Blue green ring formed Blue green ring formed

6
7 8 9 10 11

Blue green ring formed

Blue green ring formed Blue green ring formed Blue green ring formed Blue green ring formed Blue green ring formed

TEST FOR OCCULT BLOOD

INDICATIONS 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.