Physical and Chemical

Analysis of Urine
Experiment no.9
Written Report

Submitted by:
Barrameda, Suzette D.
Submitted to:

Mr. Dayle Daniel G. Sorveto

 November 18, 2019
BMLS-2B

Written Report
Activity # 9

Abstract:
Urinalysis involves the physical, chemical and microscopic analysis. This lab focused on the
physical and chemical analysis of urine. Physical urinalysis involves observing colour, odour, volume, and
clarity. There are many things that affect urine colour, including fluid balance, diet, medicines, and diseases.
The intensity of aurine specimen colour depends on its concentration. A darker colour indicates a higher
concentration. Urine colour ranges from colourless to black. Urine is normally transparent; however
bacteria, blood, sperm, crystals, mucus or other particles can make urine look cloudy. Urine clarity ranges
from clear to turbid. Urine does not smell very strong, but has a slightly "nutty" or aromatic odour. Some
diseases cause a change in the odour of urine. An E. coli bacterial infection can cause a foul odour, while
diabetes or starvation can cause a sweet, fruityodour. As the name suggests, urinalysis is the analysis of
urine. Urine is analyzed for the presence or absence of chemicals or microbes and other characteristics
which may aid in the diagnosis of disease, and screening asymptomatic populations for undetected
disorders, and monitoring the progress of disease and the effectiveness of therapy. Blood is also analyzed
in this way, however urinalysis is preferred since it is a less invasive procedure. It is therefore patient
friendly. 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: Characteristics of the urine change, depending on
influences such as water intake, exercise,
The urinary system’s ability to filter the
environmental temperature, nutrient intake, and
blood resides in about 2 to 3 million tufts of
other factors. Some of the characteristics such as
specialized capillaries—the glomeruli—
color and odor are rough descriptors of your state
distributed more or less equally between the two
of hydration. For example, if you exercise or
kidneys. Because the glomeruli filter the blood
work outside, and sweat a great deal, your urine
based mostly on particle size, large elements like
will turn darker and produce a slight odor, even if
blood cells, platelets, antibodies, and albumen are
you drink plenty of water. Athletes are often
excluded. The glomerulus is the first part of the
advised to consume water until their urine is
nephron, which then continues as a highly
clear. This is good advice; however, it takes time
specialized tubular structure responsible for
for the kidneys to process body fluids and store it
creating the final urine composition. All other
in the bladder. Another way of looking at this is
solutes, such as ions, amino acids, vitamins, and
that the quality of the urine produced is an
wastes, are filtered to create a filtrate composition
average over the time it takes to make that urine.
very similar to plasma. The glomeruli create
Producing clear urine may take only a few
about 200 liters (189 quarts) of this filtrate every
minutes if you are drinking a lot of water or
day, yet you excrete less than two liters of waste
several hours if you are working outside and not
you call urine.
drinking much. 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 inorganic substances such as
urea acid, creatine sodium chloride, ammonia,
sulfates, and phosphates as its principal
ingredients. a complete urinalysis involves an
examination of the physical characteristics of
urine, a 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 as possible. In this
experiment, physical and chemical evaluations of
urine sample will be conducted.
II. Objectives:
 Describe the characteristics of a normal
urine sample, including normal range of
pH, osmolarity, and volume.
 To test qualitatively for the presence of
some normal and abnormal organic
constituents in urine.
III. Methodology:
EQUIPMENTS
(5) 10-mL Test Tubes
Urinometer
pH paper
10-mL graduated cylinder
Hot Plate
400-mL beaker
REAGENTS
5% NaOH Bromine Water
Saturated Pieric Acid 5% NaOH
Conc. NH4OH Conc. HNO3
5% Sodium Nitropusside
Procedure
A. Collection of Urine Sample
Obtain a urine container from the hospital. As you
start to urinate, let some urine fall into the toiler,
then collect about 50-mL of urine in the container
provided. Analysis of urine should be done within
an hour after the collection.
B. Physical Characteristics of Urine
1. Color
The color of normal urine is usually light yellow
to amber. The yellow color of urine is due to the
yellow pigment, urochrome. Place 2 mL urine
sample in a test and observe the color.
2. Odor
Slightly aromatic, characteristics of freshly
voided urine. Urine becomes more ammonia-like
upon standing due to bacterial activity.
3. Turbidity
Normal urine is transparent or clear; becomes
cloudy upon standing. Cloudy urine may be
evidence of phosphates, urates, mucus, bacteria,
epithelial cells or leukocytes.
4. pH
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.
5. Specific Gravity
Measure the specific gravity if urine using the
refractometer.
1. Fill the cylinder with enough urine using
urinometer will float in the urine and not
touch the bottom.
2. Be careful not to drop the urinometer in
the cylinder. Gently release it in order not
to break or burst the cylinder. It should
not touch the sides or bottom of cylinder.
3. The specific gravity can be read on the
scale on the stern of the urinometer at the
meniscus.
4. The specific gravity of water is 1,000
with respect to temperature. The
urinometer can be checked periodically
against this standard to ensure quality
control at that temperature.
C. Chemical Analysis of Urine
1. Test for Urea
a. To 1-mL of urine, add 10 drops of 5%
NaOH and 4 drops of bromine water.
b. Observe the production of N2 gas.
2. Test for Uric acid 8. Test for Phosphates
To 1-mL of urine, add 3 drops of concentrated 1. Render 5-mL of urine basic with 2 drops
HNO3. Heat in a water bath to dryness and then of concentrated NH4OH.
add 2 drops of 1M NH4OH solution. Observe the 2. Warm and note the production of a
formation of reddish-violet color. precipitate.
3. Test for Creatinine VI. Data and Results:
To 3-mL of urine, add 1-mL of saturated picric Physical Characteristics:
acid solution and 1-mL of 5% NaOH. The
development of orange color indicates the PROPERTY
presence of creatinine. OF
TEST OBSERVATION
NORMAL
4. Test for Ketone Bodies URINE
Light Yellow
1. To 2-mL of urine, add 1-mL of 5% Light
Color and Color with
sodium nitropusside solution. Yellow /
Odor Ammoniacal
Ammoniacal
2. Mix. Smell
3. Add concentrated NH4OH slowly on the Turbidity Clear Clear
side of the tube to form a layer. Specific
4. Note the color produced at the point of 1.015 1.002-1.029
Gravity
contact. pH 6.0 4.6-7.9
5. A purple not a brown color is positive for
Chemical Characteristics:
ketone bodies.
5. Test for Bile Pigments PROPERTY
OF
Take 3-mL of concentrated HNO3 in a test tube. TEST OBSERVATION
NORMAL
Add 3-mL of urine and carefully along the side of URINE
the test tube without shaking, so that the two Urea Positive Positive
fluids do not mix. A display of colors from
Uric Acid Positive Positive
yellow, red, violet, blue, and green will be noted
at the point of contact of the two liquids, which Creatinine Positive Positive
indicates the presence of bile pigments in the Ketone
Negative Negative
urine. Bodies
Bile
6. Test for Glucose Negative Negative
Pigments
Three layer; The
a. Place 1-mL of Benedict’s reagent in a test
upper layer has a
tube and add 10 drops of urine. cloudy precipitate; Little no
b. Boil for 1-2minutes Glucose middle layer has a glucose
c. Note the change in color of the solution green precipitate; present
7. Test for Chloride bottom layer is
light blue
a. Place 5-mL of urine in a test tube and
Positive /
acidify with 2 drops of concentrated Chloride Cloudy precipitate Forms white
HNO3 precipitate
b. Add 2 drops of 0.01 M AgNO3 Positive /
c. Note what is produced. Phosphate Cloudy precipitate Forms
precipitate
V. Discussion:
In this experiment, I gathered a sample urine and
observed using physical and chemicals tests. To
determine whether there are abnormalities
present in the source of the sample. The physical
test observes the color, odor, turbidity, as well as
the pH of the urine. The sample exhibited the
physical properties of a normal urine, as it is light
yellow in color; aromatic in odor; a specific
gravity of 1.015which is in range; and a pH of 6.0.
First chemical test was for UREA, sodium
hypobromite is dropped onto a sample of urine.
This produces N2 gas which indicates the
presence of Urea. When urine is treated with
sodium Hypobromite solution containing excess
of sodium hydroxide, the carbon dioxide and
nitrogen gas are evolved. CO2 after reacting with
NaOH gives Na HCO3. Second test is for URIC
ACID, a reddish-violet color indicates the
presence of uric acid in the sample. 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. Ketone bodies is examined in the
next test using the Nitropusside reaction.
Acetoacetic acid reacts with sodium
nitroferricyanide and glycine in an alkaline
medium to produce a violet-to-purple colored
complex. 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 the Gmelin test, it produces
Different coloured rings between the two layers
are visible if bile pigments are present as they are
oxidized to various chemical products. In this
case, concentrated nitric acid is the oxidizing
agent. It did not produce the multi-colored
reaction, indicating there is little to none bilirubin
present. Test performed is the Benedict’s test
used to determine the amount of glucose that is
present in the urine. When Benedict’s solution
and simple carbohydrates are heated, the solution
changes to orange red/ brick red. A blue and
green coloration was observed indicating there is
none or small quantities of glucose present in the
urine. As for the white precipitate.
VI. Conclusion:
I therefore conclude that the based from the data
researches, gatherings, and observed qualities of
the urine sample. Routine chemical examination
of urine has changed dramatically since the first
test It is safe to say that the source of the sample
is a healthy individual.
VII. Question and Answers:
1. Determine the term and name a condition
that may cause an increase in of the
following constituents in urine.
a. Glycosuria
Glycosuria is a condition in which a person's
urine contains more sugar, or glucose, than it
should. It typically occurs due to high blood sugar
levels or kidney damage,
b. Ketonuria
Ketonuria: A condition in which abnormally
high amounts of ketones and ketone bodies (a
byproduct of the breakdown of cells) are present
in the urine.
c. Proteinuria
Protein content in urine, often due to leaky or
damaged glomeruli.
d. Hemoglobinuria
Hemoglobinuria is a condition in which
the oxygen transport protein hemoglobin is
found in abnormally high concentrations in
the urine.
e. Hematuria
The presence of blood in the urine
Urea: Uremia
Chloride: Adrenocortical Insufficiency
Sodium: Barrter Syndrome
Potassium: Hyperkalemia
Creatinine: Glomerulonephritis
 2. State an abnormal condition which
would cause the following constituents to
be present in the urine.
a. Bilirubin: Liver Cirrhosis
b. Nitrites: UTI
c. Ketone Bodies: Ketonuria
d. Leukocytes: UTI
3. State the normal values for each of the
following.
a. pH: 4.5-8.0
b. Protein: 0-20mg/dl
c. Glucose: 0-0.8mmol/dl
d. Ketone Bodies: <20 mg/dl
e. Bilirubin: 0-0.4mg/dl
f. Specific Gravity: 1.010-1.030
4. Account for the basic principle in the
formation of color for the following tests:
A. Test for chloride
The white/cloudy precipitate was formed from
adding nitric acid and silver nitrate to the urine,
which then produces a white precipitate. This
white precipitate indicates the presence of
chloride, as chloride reacts with silver nitrate.
Nitric is there to acidify the solution.
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
VII. Certification:
Barrameda, Suzette D. ________________
References:
 https://www.nps.gov/museum/publicatio
ns/conserveogram/06-03.pdf
 https://www.labce.com/spg506350_keto
ne_reaction.aspx
 https://opentextbc.ca/anatomyandphysio
logy/chapter/25-1-physical-
characteristics-of-urine/
 https://www.slideshare.net/1000004247
73559/physical-and-chemical-
examination-of-urine
 https://www.nps.gov/museum/publicatio
ns/conserveogram/06-03.pdf