ACTIVITY No.

Physical and Chemical Analysis of Urine

Members:
Cabaña, Maria Laica
Dimaano, Sheridan
Saldon, Precious
Delos Reyes, Dominic
Dimayacyac, Justin
Gavilan, John Andre

Submitted to:
Dayle Daniel Sorveto
 ABSTRACT
Urinalysis was the first laboratory test performed in medicine and has been used for several thousand years. Today
urinalysis continues to be a powerful tool in obtaining crucial information for diagnostic purposes in medicine. Urine is
an unstable fluid, and changes to its composition begin to take place as soon as it is voided. As such, collection,
storage, and handling are important issues in maintaining the integrity of this specimen. In the laboratory, urine can
be characterized by physical appearance, chemical composition, and microscopically. Physical examination of urine
includes description of color, odor, clarity, volume, and specific gravity. Chemical examination of urine includes the
identification of protein, blood cells, glucose, pH, bilirubin, urobilinogen, ketone bodies, nitrites, and leukocyte
esterase. Finally, microscopic examination entails the detection of crystals, cells, casts, and microorganisms.
I. Introduction pH paper
10-mL graduated cylinder
Urinalysis includes description of color, odor, clarity, Hot Plate
volume, and specific gravity. Chemical examination of 400-mL beaker
urine includes the identification of protein, blood cells,
glucose, pH, bilirubin, urobilinogen, ketone bodies,
nitrites, and leukocyte esterase. REAGENTS
5% NaOH Bromine Water
The major function of the urinary system is to remove Saturated Pieric Acid 5% NaOH
waster products from out bodies and help regulate the Conc. NH4OH Conc. HNO3
volume and composition of blood by the production 5% Sodium Nitropusside
and excretion of urine. The composition of urine can
vary greatly and constantly fluctuates with dietary
intake (food and water) and metabolic activity. Urine
consists mostly of water with various organic and
Obtain a urine container from the hospital.
inorganic substances such as urea acid, creatine
As you start to urinate, let some urine fall
sodium chloride, ammonia, sulfates, and phosphates
into the toiler, then collect about 50-mL of
as its principal ingredients. A clinical examination of
urine in the container provided. Analysis of
urine can provide a convenient, cost effective and
urine should be done within an hour after
non-invasive means of assessing kidney function and
the collection
providing and overall assessment of our body’s
health.
Procedure
Typically, a complete urinalysis involves an
examination of the physical characteristics of urine, a A. Collection of Urine Sample
chemical analysis and a microscopic examination of
urine sediment. Urine should be collected in a clean
container, stored in a cool place, and tested as soon The color of normal urine is usually light
as possible. In this experiment, physical and chemical yellow to amber. The yellow color of urine
evaluations of urine sample will be conducted. is due to the yellow pigment, urochrome.

II. Methodology B. Physical Characteristics of Urine

EQUIPMENTS 1. Color
(5) 10-mL Test Tubes
Urinometer 2. Odor
 Slightly aromatic, characteristics of freshly
voided
Cover urine. Shake
the crystal. Urinewellbecomes more
from side to
sideammonia-like
to even out theuponurine. standing due to
bacterial activity.

Look into the eyepiece and record the

results.

Take note of the smell of urine C. Chemical Analysis of Urine

1.
Test
3. Turbidity To 1-mL of urine, add 10 drops of 5% for
NaOH and 4 drops of bromine water. Urea

Normal urine is transparent or clear;

becomes cloudy upon standing. Cloudy
urine may be evidence of phosphates,
urates, mucus, bacteria, epithelial cells or
leukocytes.
Observe the production of N2 gas

4. pH
2. Uric Acid

Normal pH range of urine is 4.5-8.0

Determine the pH of urine by dipping a
strip of pH paper into the test tube
To 1-mL of urine, add 3 drops of
concentrated HNO3. Heat in a water
bath to dryness and then add 2 drops
of 1M NH4OH solution. Observe the
formation of reddish-violet color.

Measure the specific gravity if urine using

the refractometer. 3. Test for Creatinine

5. To 3-mL of urine, add 1-mL of saturated

Spe picric acid solution and 1-mL of 5% NaOH.
cific The development of orange color
Grav indicates the presence of creatinine
ity
4. Test for Ketone Bodies
Place 2 drops of urine onto the measuring
prism To 2-mL of urine, add 1-mL of 5%
sodium nitropusside solution
 7. Test for Chloride
Mix

Place 5-mL of urine in a test tube and

acidify with 2 drops of concentrated HNO3
Add concentrated NH4OH slowly on the
side of the tube to form a layer.
Add 2 drops of 0.01 M AgNO3

Note what is produced.

Note the color produced at the point of

contact. 8. Test for Phosphates

Render 5-mL of urine basic with 2

drops of concentrated NH4OH
A purple not a brown color is positive for
ketone bodies.

Warm and note the production of a

5. Test for Bile Pigments.
precipitate.

Take 3-mL of concentrated HNO3 in a test

tube. Add 3-mL of urine and carefully III. Data and Results
along the side of the test tube without
Physical Characteristics
shaking, so that the two fluids do not mix.
A display of colors from yellow, red, violet,
PROPERTY OF
blue, and green will be noted at the point TEST OBSERVATION
NORMAL URINE
of contact of the two liquids, which
indicates the presence of bile pigments in Light Yellow
the urine. Color with Light Yellow /
Color and Odor
Ammoniacal Ammoniacal
Smell
6. Test for Glucose
Turbidity Clear Clear
Place 1-mL of Benedict’s reagent in a test
tube and add 10 drops of urine. Specific Gravity 1.018 1.002-1.030

Boil for 1-2minutes pH 6.0 4.5-8.0

Note the change in color of the solution Chemical Characteristics

 PROPERTY carbon dioxide and nitrogen gas are evolved. CO2
TEST OBSERVATION OF NORMAL after reacting with NaOH gives Na HCO3.
URINE
The second test is for URIC ACID, a reddish-violet
Urea Positive Positive color indicates the presence of uric acid in the
sample. The principle of forming a red violet dye is
Uric Acid Positive Positive due to the ammonium salts or murexide. These salts
are formed when nitric acid is added to a sample
Creatinine Positive Positive
urine, which then evaporates it. And when the residue
Ketone is added with NH4OH and forms a reddish color which
Negative Negative
Bodies is the murexide, it indicates that uric acid is present.
Bile
Negative Negative The next test is determining the presence of
Pigments
Has 3 layer; upper creatinine in the urine by the use of picric acid
layer has a cloudy reaction or the Jaffe reaction. An orange coloration is
Little to no
precipitate; middle formed when creatinine reacts with the picrate ion
Glucose glucose
layer has a green present in the urine.
present
precipitate; bottom
layer is blue Ketone bodies is examined in the next test using the
Positive / nitropusside reaction. Acetoacetic acid reacts with
Chloride Cloudy precipitate Forms white sodium nitroferricyanide and glycine in an alkaline
precipitate medium to produce a violet-to-purple colored
Positive / complex.
Phosphate Cloudy precipitate Forms
precipitate Next test is for the examination of bile pigments in the
urine. Or rather to determine whether there is bilirubin
present in the urine sample. The test used is called
IV. Discussion the Gmelin test, it produces Different coloured rings
between the two layers are visible if bile pigments are
In this experiment, a sample urine was gathered and
present as they are oxidised to various chemical
is observed using physical and chemicals tests. To
products. In this case, concentrated nitric acid is the
see whether there are abnormalities present in the
oxidizing agent. It did not produce the multi-colored
source of the sample.
reaction, indicating there is little to none bilirubin
The physical test observes the color, odor, turbidity, present.
as well as the pH of the urine. The sample exhibited
the physical properties of a normal urine, as it is light The next test performed is the benedict’s test used to
determine the amount of glucose that is present in the
yellow in color; aromatic in odor; a specific gravity of
1.018 which is in range; and a pH of 6.0 which is also urine. When Benedict’s solution and simple
carbohydrates are heated, the solution changes to
in range.
orange red/ brick red. This reaction is caused by the
The chemical tests observes the sample urine using reducing property of simple carbohydrates. The
chemical reagents, to observe the presence of certain copper (II) ions in the Benedict’s solution are reduced
chemicals. to Copper (I) ions, which causes the color change. A
blue and green coloration was observed indicating
The first chemical test was for UREA, sodium
there is none or small quantities of glucose present in
hypobromite is dropped onto a sample of urine. This
the urine. As for the white precipitate, Benedict’s
produces N2 gas which indicates the presence of
quantitative reagent contains potassium thiocyanate
Urea. When urine is treated with sodium hypobromite
and is used to determine how much reducing sugar is
solution containing excess of sodium hydroxide, the
present. This solution forms a copper thiocyanate Hemoglobinuria is a condition in which
precipitate which is white. the oxygen transport protein hemoglobin is found in
abnormally high concentrations in the urine
The second to the last test is a test for presence of
chloride. The white/cloudy precipitate was formed e. Hematuria
from adding nitric acid and silver nitrate to the urine,
The presence of blood in the urine
which then produces a white precipitate. This white
precipitate indicates the presence of chloride, as Urea: Uremia
chloride reacts with silver nitrate. Nitric is there to
acidify the solution. Chloride: Adrenocortical Insufficiency

The last test was for determining phosphate in the Sodium: Barrter Syndrome
sample. Ammonium hydroxide reacts with a sufficient Potassium: Hyperkalemia
concentration of phosphate ions, forming a
precipitate. Indicating the presence of phosphate. Creatinine: Glomerulonephritis

V. Conclusion 2. State an abnormal condition which would cause the

following constituents to be present in the urine.
Based from the data gatherings, researches, and
observed qualities of the urine sample. It is safe to a. Bilirubin: Liver Cirrhosis
say that the source of the sample is a healthy
b. Nitrites: UTI
individual. As the urine provided by the benefactor
has exhibited what a normal urine has, both c. Ketone Bodies: Ketonuria
physically and chemically.
d. Leukocytes: UTI
VI. Question and Answer
3. State the normal values for each of the following
1. Determine the term and name a condition that may
cause an increase in of the following constituents in a. pH: 4.5-8.0
urine. b. Protein: 0-20mg/dL
a. Glycosuria c. Glucose: 0-0.8mmol/dL
Glycosuria is a condition in which a person's urine d. Ketone Bodies: <20 mg/dL
contains more sugar, or glucose, than it should. It
typically occurs due to high blood sugar levels or e. Bilirubin: 0-0.4mg/dL
kidney damage.
f. Specific Gravity: 1.010-1.030
b. Ketonuria
4. Account for the basic principle in the formation of
Ketonuria: A condition in which abnormally high color for the following tests:
amounts of ketones and keytone bodies (a byproduct
a. test for chloride
of the breakdown of cells) are present in the urine.
The white/cloudy precipitate was formed from adding
c. Proteinuria
nitric acid and silver nitrate to the urine, which then
Protein content in urine, often due to leaky or produces a white precipitate. This white precipitate
damaged glomeruli indicates the presence of chloride, as chloride reacts
with silver nitrate. Nitric is there to acidify the solution.
d. Hemoglobinuria
b. test for phosphate
Ammonium hydroxide reacts with a sufficient
concentration of phosphate ions, forming a
precipitate. Indicating the presence of phosphate
c. test for glucose
When Benedict’s solution and simple carbohydrates
are heated, the solution changes to orange red/ brick
red. This reaction is caused by the reducing property
of simple carbohydrates. The copper (II) ions in the
Benedict’s solution are reduced to Copper (I) ions,
which causes the color change
d. test for creatinine
An orange coloration is formed when creatinine reacts
with the picrate ion present in the urine.
e. test for uric acid
The principle of forming a red violet dye is due to the
ammonium salts or murexide. These salts are formed
when nitric acid is added to a sample urine, which
then evaporates it. And when the residue is added
with NH4OH and forms a reddish color which is the
murexide, it indicates that uric acid is present
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References:
Photo Documentation

Positive result for ketone bodies

Specific gravity of the sample
 Chloride test result

Urea test result

Colorimetric Test Results

Creatinine Test Result

Test Results