DEPARTMENT OF CHEMISTRY

College of Arts and Sciences

CENTRAL PHILIPPINE UNIVERSITY
ILOILO CITY

CHEMISTRY_5001__ SCORE & TEACHER’S SIGNATURE ______________________

GROUP NO. ___2___ SCHEDULE __________________ DATE _________________

MEMBERS: 1. Eriquez, Micha Ella 4. Tambuson, Kynt

Nathaniel

2. Rocero, Lloyd Andre 5. Ysug, Erwin

3. Suravilla, Kria Faye 6.

6 – NUCLEIC ACIDS
EXPERIMENT NO. & TITLE _____________________________________________

I. OBJECTIVES
 To be able to isolate DNA either thymus or liver
 To be able to isolate RNA from yeast
 To be able to detect qualitatively for the presence of simple components of hydrolyzed
DNA and RNA, respectively
II. INTRODUCTION
The two types of nucleic acids are DNA (deoxyribonucleic acid) and RNA (ribonucleic
acid). DNA carries genetic information and is typically found in the nucleus of cells. It consists of
two strands twisted into a double helix structure, made up of nucleotides containing a sugar-
phosphate backbone and nitrogenous bases (adenine, thymine, cytosine, and guanine). RNA is
involved in various cellular processes, including protein synthesis. It is single-stranded and
contains ribose sugar instead of deoxyribose found in DNA. RNA also contains four nitrogenous
bases: adenine, uracil, cytosine, and guanine. There are several types of RNA, including
messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA), each with specific
functions in protein synthesis.
The hydrolysis reaction of DNA and RNA involves breaking the phosphodiester bonds
between nucleotides, resulting in the separation of the nucleic acid strands into individual
nucleotides. In DNA, the hydrolysis reaction typically involves the addition of water molecules to
the phosphodiester bonds between adjacent nucleotides. This process is catalyzed by enzymes
known as nucleases. As a result of hydrolysis, the DNA strand is cleaved into its component
nucleotides, each containing a phosphate group, a deoxyribose sugar, and a nitrogenous
base.Similarly, in RNA, hydrolysis breaks the phosphodiester bonds between ribonucleotides.
This reaction also requires the addition of water molecules and is catalyzed by nucleases. As a
result, the RNA molecule is fragmented into individual ribonucleotides, each consisting of a
phosphate group, a ribose sugar, and a nitrogenous base.Overall, the hydrolysis of DNA and
RNA plays a crucial role in various biological processes, including DNA replication, RNA
degradation, and gene expression..
Benedict's Test is used to detect the presence of reducing sugars, such as glucose and
fructose, which have free aldehyde or ketone groups. In the test, Benedict's reagent (which
contains copper(II) sulfate, sodium citrate, and sodium carbonate) is added to the solution
containing the sugar and heated. If a reducing sugar is present, it will reduce the blue copper(II)
ions in Benedict's reagent to form a brick-red precipitate of copper(I) oxide. While Orcinol Test is
specifically used to detect the presence of pentoses, particularly ribose and deoxyribose sugars.
In the Orcinol test, the sample is treated with orcinol reagent (containing orcinol, sulfuric acid,
and ferric chloride) and heated. If pentoses are present, they react with the orcinol reagent to
produce a bluish-green color. Test for Purine Bases, such as adenine and guanine, can be
detected using various methods. One common method involves the use of a reagent called
trichloroacetic acid (TCA). When TCA is added to a solution containing purine bases, it
precipitates the purines as white crystals, which can then be collected and identified. Lastly, the
test for Inorganic Phosphate can be detected using the molybdate blue method. In this test, a
sample containing phosphate ions is treated with ammonium molybdate and sulfuric acid,
followed by the addition of ascorbic acid. If phosphate ions are present, they react with the
molybdate reagent to form a blue-colored complex, known as molybdenum blue.These tests are
essential in biochemical analysis for detecting specific components in a sample, such as sugars,
nucleic acids, or phosphate ions.
 DEPARTMENT OF CHEMISTRY
College of Arts and Sciences
CENTRAL PHILIPPINE UNIVERSITY
ILOILO CITY

CHEMISTRY_5001__ SCORE & TEACHER’S SIGNATURE ______________________

GROUP NO. ___2___ SCHEDULE __________________ DATE _________________

MEMBERS: 1. Eriquez, Micha Ella 4. Tambuson, Kynt

Nathaniel

2. Rocero, Lloyd Andre 5. Ysug, Erwin

3. Suravilla, Kria Faye 6.

EXPERIMENT NO. & TITLE _____________________________________________

III. RESULTS AND DISCUSSION

A. MICROSCOPIC ANALYSIS.
Search through the internet the microscopic structure of DNA precipitate and then draw.

DNA
Describe the appearance of the precipitate: The DNA precipitate appears as clumped or
aggregated white material in the solution. When DNA is precipitated using isopropanol or
ethanol in the presence of a high-concentration salt solution, the DNA molecules clump together
and form a visible white precipitate. This clumped DNA precipitate can be observed as a solid
mass that separates from the solution during the precipitation process. The DNA precipitate is
typically collected by centrifugation, where it condenses into a pellet at the bottom of the tube.
The pellet contains the relatively pure DNA, while the supernatant contains the excess salt and
other cellular debris that were removed during the precipitation process.

B. QUALITATIVE TESTS FOR HYDROLYZED DNA

Test for OBSERVATIONS RESULT
DNA +/-
Reducing sugar no color change -
(Benedict’s test)
Pentoses formation of deep blue coloration +
(Orcinol test)
Purine bases formation of a white or cream-colored +
precipitate.
Inorganic phosphate formation of yellow precipitate +

DISCUSSION:
1. Should all your results for the tests be positive? Why or why not?
Not all tests are positive because when we hydrolyze DNA, we're essentially breaking
down the molecule into its constituent parts. This can be done through various methods, such
 DEPARTMENT OF CHEMISTRY
College of Arts and Sciences
CENTRAL PHILIPPINE UNIVERSITY
ILOILO CITY

CHEMISTRY_5001__ SCORE & TEACHER’S SIGNATURE ______________________

GROUP NO. ___2___ SCHEDULE __________________ DATE _________________

MEMBERS: 1. Eriquez, Micha Ella 4. Tambuson, Kynt

Nathaniel

2. Rocero, Lloyd Andre 5. Ysug, Erwin

3. Suravilla, Kria Faye 6.

EXPERIMENT NO. & TITLE _____________________________________________

as acid hydrolysis or enzymatic digestion. Once we've broken down the DNA, we can use
various tests to identify the different components that are present. One of the first tests we can
do is Benedict's test for reducing sugars. This test is used to detect the presence of reducing
sugars, such as glucose. However, when we hydrolyze DNA, we don't typically get reduced
sugars like glucose. So, the result of Benedict's test is negative (-).

Next, we can use the Orcinol test to detect the presence of pentoses. Pentoses are a
type of sugar that are found in DNA, specifically the pentose sugar 2-deoxyribose. The Orcinol
test is specific for pentoses, so when we run the test on our hydrolyzed DNA sample, we get a
positive (+) result. We can also use tests to detect the presence of purine bases. Purine bases
are a type of nitrogenous base that are found in DNA, specifically adenine and guanine. When
we hydrolyze DNA, we release these purine bases, so the test for purine bases will be positive
(+). Finally, we can use tests to detect the presence of inorganic phosphate. Inorganic
phosphate is a byproduct of the hydrolysis reaction, so when we run the test, we get a positive
(+) result.

Overall, these tests help us identify the different components that are present in our
hydrolyzed DNA sample. By using these tests, we can gain a better understanding of the
structure and composition of DNA.

B. MICROSCOPIC ANALYSIS.
Search through the internet the microscopic structure of RNA precipitate and then draw.

RNA

Describe the appearance of the precipitate: The microscopic structure of RNA

precipitate, as revealed by cryo-EM, appears as anisotropic ensembles of single-stranded RNA
molecules with branched secondary structures and varying shapes and sizes. The phosphorus-
rich RNA backbone is visible, and the use of staining agents like thionine can enhance the
contrast and provide insights into the location of mRNA within complex structures like LNPs.
 DEPARTMENT OF CHEMISTRY
College of Arts and Sciences
CENTRAL PHILIPPINE UNIVERSITY
ILOILO CITY

CHEMISTRY_5001__ SCORE & TEACHER’S SIGNATURE ______________________

GROUP NO. ___2___ SCHEDULE __________________ DATE _________________

MEMBERS: 1. Eriquez, Micha Ella 4. Tambuson, Kynt

Nathaniel

2. Rocero, Lloyd Andre 5. Ysug, Erwin

3. Suravilla, Kria Faye 6.

EXPERIMENT NO. & TITLE _____________________________________________

B. QUALITATIVE TESTS FOR HYDROLYZED RNA

Test for OBSERVATIONS RESULT
RNA +/-
Reducing sugar from red litmus paper to blue litmus paper +
(Benedict’s test)
Pentoses After boiling in a water bath the color will change from yellow- -
(Orcinol test) orange to dark green.
Purine bases White cloudy precipitate formed +
Inorganic The appearance formed yellow cloudy precipitate. +
phosphate

DISCUSSION:
1. Should all your results for the tests be positive? Why or why not?
The results of the qualitative tests on hydrolyzed RNA should not all be positive because each
test targets a specific component that may or may not be present in the sample. The tests are
designed to detect the presence or absence of reducing sugars, pentoses, purine bases, and
inorganic phosphate, which are the products of RNA hydrolysis. Since not all components are
always present in detectable amounts, the results will vary, with some tests giving positive
results and others giving negative results.
IV. SUMMARY AND GENERALIZATIONS
In summary, the objectives of this lab experiment—which were to comprehend the basic
characteristics of nucleic acids, more especially DNA and RNA—were successfully met.
Participant familiarity with the extraction methods essential for molecular biology research was
obtained by isolating DNA from thymus and liver samples as well as RNA from yeast.

Additionally, the qualitative identification of basic hydrolyzed DNA and RNA components
revealed important details about the molecular makeup of these nucleic acids. This part of the
experiment highlighted the value of meticulous experimental design and interpretation in
molecular biology research in addition to reinforcing theoretical understanding.
All things considered, this lab exercise opened the door for additional research and
experimentation in the subject by providing a hands-on introduction to the ideas and techniques
essential to the study of nucleic acids.

V. REFERENCE

7 – BILE

I. OBJECTIVES
 DEPARTMENT OF CHEMISTRY
College of Arts and Sciences
CENTRAL PHILIPPINE UNIVERSITY
ILOILO CITY

CHEMISTRY_5001__ SCORE & TEACHER’S SIGNATURE ______________________

GROUP NO. ___2___ SCHEDULE __________________ DATE _________________

MEMBERS: 1. Eriquez, Micha Ella 4. Tambuson, Kynt

Nathaniel

2. Rocero, Lloyd Andre 5. Ysug, Erwin

3. Suravilla, Kria Faye 6.

EXPERIMENT NO. & TITLE _____________________________________________

● To be able to test for the presence of bile components and describe their reactions
● To be able to isolate cholesterol from bile and describe its reactions to chemical tests
II. INTRODUCTION
Bile is a digestive fluid produced by the liver and stored in the gallbladder. It plays a
crucial role in the digestion and absorption of fats and fat-soluble vitamins in the small intestine.
Bile is primarily composed of water, bile salts, bilirubin, cholesterol, electrolytes, and
phospholipids. Bile salts are the most abundant organic component and are essential for the
emulsification and digestion of fats.Bile aids in the digestion and absorption of fats by
emulsifying them into smaller droplets, which increases the surface area for the action of
digestive enzymes, such as lipase. This process enhances the efficiency of fat digestion and
allows for the absorption of fatty acids and fat-soluble vitamins (A, D, E, and K) in the small
intestine. Bile salts have both hydrophilic (water-attracting) and hydrophobic (water-repelling)
regions, which enable them to surround fat molecules and form micelles. These micelles
disperse fat droplets into smaller particles, making them more accessible to lipase enzymes for
digestion. Bile is continuously produced by the liver and stored in the gallbladder between
meals. When food, especially fatty food, enters the small intestine, hormonal signals stimulate
the gallbladder to contract and release bile into the duodenum (the first part of the small
intestine) through the common bile duct. The production and release of bile are regulated by
various factors, including neural signals, hormonal signals (such as cholecystokinin), and the
presence of fatty acids and amino acids in the small intestine. Overall, bile plays a crucial role in
the digestion and absorption of fats and fat-soluble vitamins, contributing to overall nutrient
absorption and metabolism in the body.
Bile consists of water, electrolytes, bile acids, cholesterol, phospholipids, and bilirubin.
Proteins found in bile include albumin, globulins, and enzymes such as alkaline phosphatase.
Lipids include cholesterol, phospholipids, and fatty acids. Inorganic constituents include
electrolytes such as sodium, potassium, chloride, bicarbonate, and bilirubin.
III. RESULTS AND DISCUSSION
A. REACTION OF BILE

Red Litmus Paper Blue Litmus Paper Congo Red Paper Phenolphthalein

Observations It retained red that It retained blue that It retained red that remained as
means it’s acidic yellow
means it’s basic. means it’s acidic.

What is the pH of your bile sample? (Acidic, basic or neutral)

Based on the result of the pH paper the pH of the bile sample was 7, therefore the pH of
the bile sample is neutral.

Normally, what is the pH value of bile? Is your sample within the normal limit?
 DEPARTMENT OF CHEMISTRY
College of Arts and Sciences
CENTRAL PHILIPPINE UNIVERSITY
ILOILO CITY

CHEMISTRY_5001__ SCORE & TEACHER’S SIGNATURE ______________________

GROUP NO. ___2___ SCHEDULE __________________ DATE _________________

MEMBERS: 1. Eriquez, Micha Ella 4. Tambuson, Kynt

Nathaniel

2. Rocero, Lloyd Andre 5. Ysug, Erwin

3. Suravilla, Kria Faye 6.

EXPERIMENT NO. & TITLE _____________________________________________

Normally, bile is composed of bile acids and salts, phospholipids, cholesterol, pigments,
water, and electrolyte chemicals that keep the total solution slightly alkaline (with a pH of about
7 to 8). Therefore, the normal pH range for bile is approximately 7 to 8, which is slightly alkaline,
and the pH our bile sample is 7 which still fits and with in the normal limit.
B. NUCLEOPROTEINS AND BILE ACIDS
Observation:
After adding acetic acid to the bile sample it looks like the bile precipitated out of the
solution since the solution becomes clear and the precipitate is yellow which is the color of the
bile sample before adding the acetic acid.
What substance is produced?
Insoluble bile acid and salts.
C. INORGANIC CONSTITUENTS

Test For Observations +/-

1. Chloride ions formation of white precipitate +

2. Sulfate ions no changes in color -
3. Phosphate ions no changes in color -

D. TESTS FOR BILE PIGMENTS

Test Observations +/-

1. Gmelin’s test Shows change in color into blue and green +

2. Rosenbach’s test Shows a change in color from turbid yellow to light pink +

E. BILE ACIDS
1. Pettenkofer Test
Observation:
Turbid yellow color can be first observed on the sample and turned into cherry red color
when mixed with sulfuric acid thus it is positive for pettenkofer test.

2. Effect on Surface Tension

 DEPARTMENT OF CHEMISTRY
College of Arts and Sciences
CENTRAL PHILIPPINE UNIVERSITY
ILOILO CITY

CHEMISTRY_5001__ SCORE & TEACHER’S SIGNATURE ______________________

GROUP NO. ___2___ SCHEDULE __________________ DATE _________________

MEMBERS: 1. Eriquez, Micha Ella 4. Tambuson, Kynt

Nathaniel

2. Rocero, Lloyd Andre 5. Ysug, Erwin

3. Suravilla, Kria Faye 6.

EXPERIMENT NO. & TITLE _____________________________________________

Observation: The Pettenkofer test is not directly related to surface tension observation.
It's a chemical test used to detect the presence of bile acids, particularly bile acids like cholic
acid and chenodeoxycholic acid. It relies on the formation of a cherry-red color when bile acids
react with sulfuric acid and cane sugar. Surface tension observation would involve a different
experimental setup or test method.
What happened with the sulfur powder? Explain why.

In the Pettenkofer test for bile acids, sulfur powder is used to aid in the detection
process. When bile acids react with sulfuric acid and cane sugar, they form a complex mixture
that contains furfurals. These furfurals react with the sulfur powder to produce a cherry-red
color, indicating the presence of bile acids. Therefore, the sulfur powder acts as a catalyst in the
reaction, facilitating the formation of the characteristic color change.
F. CHOLESTEROL IN BILE

TESTS Observation +/-

Lieberman-Buchard Green Solution +

Salkowski Deep yellow or reddish-brown ring +

What is the danger of excessive amount of cholesterol in bile? Explain why.

If the bile contains too much cholesterol, the excess forms crystals, and hard stones in
the gallbladder. Gallstones are small, hard crystalline mass that forms abnormally in the
gallbladder or bile ducts from bile pigments, cholesterol, and calcium salts. Gallstones can
cause severe pain and bile duct blockage. Maintaining a healthy cholesterol level through
recommended blood tests andlowering the risk of heart disease will help improve the overall
quality of life.
IV. CONCLUSION
The bile sample was found to be neutral, with a pH of 7, which is within the normal
slightly alkaline range of 7-8 for bile due to its composition of bile acids, salts, phospholipids,
cholesterol, pigments, water, and electrolytes. The experiment tested for various bile
components, including positive results for inorganic chloride ions, bile pigments detected
through Gmelin's and Rosenbach's tests, and the presence of bile acids confirmed by the
Pettenkofer test. Additionally, the precipitation of insoluble bile acids and salts was observed
upon adding acetic acid to the bile sample. The Lieberman-Buchard and Salkowski tests were
positive for cholesterol in the bile, and excessive cholesterol in bile can lead to the formation of
gallstones, which can cause severe pain and bile duct blockage, highlighting the importance of
maintaining a healthy cholesterol level to prevent such complications.
V. REFERENCE
8 - URINE

I. OBJECTIVES
 DEPARTMENT OF CHEMISTRY
College of Arts and Sciences
CENTRAL PHILIPPINE UNIVERSITY
ILOILO CITY

CHEMISTRY_5001__ SCORE & TEACHER’S SIGNATURE ______________________

GROUP NO. ___2___ SCHEDULE __________________ DATE _________________

MEMBERS: 1. Eriquez, Micha Ella 4. Tambuson, Kynt

Nathaniel

2. Rocero, Lloyd Andre 5. Ysug, Erwin

3. Suravilla, Kria Faye 6.

EXPERIMENT NO. & TITLE _____________________________________________

 To be able to isolate and microscopically examine some urine components such as

hippuric acid, uric acid, calcium sulfate, and triple phosphate.
 To be able to test and describe for the presence of physiological inorganic and organic
constituents as well as pathological constituents of urine.
II. INTRODUCTION
Urine is a liquid waste product produced by the kidneys as they filter excess water, salts,
and waste products from the bloodstream. Normal metabolic products present in urine include
urea, creatinine, uric acid, and various electrolytes like sodium, potassium, and chloride.
Urinalysis can provide valuable information about various diseases and conditions.
Some of the conditions that can be determined through urinalysis include urinary tract
infections, kidney diseases, diabetes, liver diseases, dehydration, and certain metabolic
disorders. Abnormalities in the appearance, pH, specific gravity, protein, glucose, ketones,
blood cells, and bacteria in the urine can indicate underlying health issues.
Benedict’s test is used to detect reducing sugars, particularly glucose. Its positive result
is a formation of a brick-red precipitate. Next is the Jaffe’s test which detects creatinine, a waste
product of muscle metabolism. Its positive result is a development of a deep orange or red
color. Followed by Obermeyer’s test which detects acetone, a ketone body. Its positive result is
a formation of a purple or violet color. Then Rothera’s Nitroprusside test which detects ketone
bodies, mainly acetoacetate. Its positive result is a formation of a purple precipitate or color
change. Then the Heller’s test which detects proteins in urine. Its positive result is a formation of
a white precipitate at the junction of the urine and acid. Then Robert’s test detects urobilinogen,
a breakdown product of bilirubin. The positive result is a formation of a pink or red color. Then
the benzidine test detects blood in urine. The positive result is a formation of a blue color. Then
the murexide test detects uric acid. The positive result is a formation of a purple color. Also,
Lucke’s test detects the presence of bile salts. The positive result is a formation of a pink or red
color. Lastly, Nylander's Test detects reducing sugars, especially pentoses. Positive result is a
formation of a black precipitate or color change to gray or black.

III. RESULTS AND DISCUSSION

A. Physical Properties of Urine
Blue
Color Odor Transparency Red Litmus pH
Litmus
Observation Yello Aromatic Clear Turns blue Remains 7 pH
w odor blue color

Answer the following questions based on research.

1. What substances are responsible for the normal color of urine?
The normal color of urine is primarily due to the presence of urochrome, a pigment
resulting from the breakdown of hemoglobin. Other factors influencing urine color include the
concentration of waste products, hydration levels, and dietary intake. Generally, a pale yellow to
straw-yellow color is considered normal for urine.
2. What happens when urine is allowed to stand for some time exposed to air?
 DEPARTMENT OF CHEMISTRY
College of Arts and Sciences
CENTRAL PHILIPPINE UNIVERSITY
ILOILO CITY

CHEMISTRY_5001__ SCORE & TEACHER’S SIGNATURE ______________________

GROUP NO. ___2___ SCHEDULE __________________ DATE _________________

MEMBERS: 1. Eriquez, Micha Ella 4. Tambuson, Kynt

Nathaniel

2. Rocero, Lloyd Andre 5. Ysug, Erwin

3. Suravilla, Kria Faye 6.

EXPERIMENT NO. & TITLE _____________________________________________

When urine is allowed to stand exposed to air for some time, several changes can occur.
First urine develops an ammonia-like odor due to the breakdown of urea into ammonia by
bacterial action. Then urine may darken in color due to oxidation of certain compounds. For
example, urochrome may oxidize, causing the urine to turn more brownish. Substance like
urates, phosphates, and crystals may precipitate out of the urine, forming sediments at the
bottom of the container. Bacteria present in urine can multiply, leading to the formation of cloudy
urine. Overall, these changes are mostly due to microbial action and oxidation processes.
3. What is the normal reaction (pH) of urine when freshly voided? To what substances is
this reaction due to?
The normal pH of freshly voided urine typically ranges from about 4.6 to 8.0, but it
commonly falls between 5.0 and 6.0 for most individuals. This acidity or alkalinity is due to the
presence of various substances in urine, including acidic substances such as uric acid and
ammonium chloride, which contribute to urine acidity. Then alkaline substances such as
bicarbonate ions, which can buffer acids and contribute to urine alkalinity. Factor such as diet,
hydration level, and certain medications can influence the pH of urine.
4. What happens when urine is allowed to stand without a preservative? Why?
When urine is allowed to stand without a preservative, several changes occur due to
bacterial action and chemical reactions. Bacteria present in urine can multiply rapidly in a warm,
moist environment, leading to the decomposition of urea into ammonia. This bacterial growth
can cause a foul odor and may also lead to the formation of sediments. Various substances
present in urine, such as urates, phosphates, and crystals, can precipitate out of solution over
time. This can result in the formation of sediments or crystals at the bottom of the container. As
bacterial activity increases, the pH of urine may become more alkaline due to the conversion of
urea to ammonia by bacteria. Some compounds in urine may undergo oxidation or other
chemical reactions when exposed to air, leading to changes in color and odor. Overall, allowing
urine to stand without a preservative can lead to microbial contamination, changes in pH, and
the formation of sediments, which can affect the reliability of urine testing and analysis.
Organic and Inorganic Constituents
1. Uric Acid
Test for Observation
Murexide Test The residue turns reddish- violet
Silver reduction After putting on the strip filter paper, the filter paper become black in
test color.
Benedict’s test The color of the solution turns blue, it means uric acid is present in urine

What is uric acid? Is it normal for the urine to contain uric acid?

Uric acid is a waste product that forms when the body breaks down purines, which are
natural substances found in the body and certain foods. It is a normal body waste product that is
usually dissolved in the blood and eliminated through urine. Uric acid levels can increase when
the body produces too much or when the kidneys are not functioning properly. While having
some uric acid in the blood and urine is normal, high levels of uric acid in the urine can indicate
health conditions like gout or kidney stones. Therefore, it is normal for urine to contain uric acid,
but abnormal levels may indicate underlying health issues.

2. Indican
 DEPARTMENT OF CHEMISTRY
College of Arts and Sciences
CENTRAL PHILIPPINE UNIVERSITY
ILOILO CITY

CHEMISTRY_5001__ SCORE & TEACHER’S SIGNATURE ______________________

GROUP NO. ___2___ SCHEDULE __________________ DATE _________________

MEMBERS: 1. Eriquez, Micha Ella 4. Tambuson, Kynt

Nathaniel

2. Rocero, Lloyd Andre 5. Ysug, Erwin

3. Suravilla, Kria Faye 6.

EXPERIMENT NO. & TITLE _____________________________________________

Test for Observation

Jaffe’s test The result is indigo blue color
Obermeyer’s Test The result is indigo colored compound

What is Indican? Is it normal for the urine to contain indican?

Indican is a compound formed when the amino acid tryptophan is broken down by gut
bacteria. It is normally present in urine at low levels, around 0.13-0.21 mmol/L (32.5-52.5 mg/L).
Elevated indican above 0.4 mmol/L (100 mg/L) may suggest digestive issues or bacterial
overgrowth. While its presence is normal, abnormal indican levels warrant further medical
investigation.

3. Hippuric Acid
Test for Observation
Luke’s reaction The odor is like a shoe polish
Sublimation Hippuric acid is sublime in the side of test tube

What is Hippuric acid? Is it normal for the urine to contain Hippuric acid?
Hippuric acid is a compound that is normally present in human urine. It is formed when
benzoic acid, produced during the breakdown of certain amino acids and found in some foods
and medications, is conjugated with glycine in the liver. Yes, it is normal for urine to contain
hippuric acid. The typical excretion of hippuric acid in non-exposed individuals ranges from 0.18
to 1.0 g/g creatinine. Elevated levels may indicate exposure to certain chemicals like toluene,
but its presence in urine is considered normal.
4. Detection of pigments
Observation
Ammoniacal Zinc The result is greenish fluorescence, that means urobilin is present.
Chloride test

What are the pigments normally present in urine?

The pigments normally present in urine are urochrome or urobilin. Urochrome is the
primary pigment responsible for the yellow color of urine, while urobilin contributes to the color
variations in urine, especially in cases of liver dysfunction or increased bilirubin production.
5. Detection of Purine Bases
What are the purine bases normally present in urine?

The purine bases normally present in urine are adenine and guanine. These purine
bases are essential components of nucleic acids and are excreted in urine as part of normal
metabolic processes.

Inorganic Constituents
1. Sulfates
 DEPARTMENT OF CHEMISTRY
College of Arts and Sciences
CENTRAL PHILIPPINE UNIVERSITY
ILOILO CITY

CHEMISTRY_5001__ SCORE & TEACHER’S SIGNATURE ______________________

GROUP NO. ___2___ SCHEDULE __________________ DATE _________________

MEMBERS: 1. Eriquez, Micha Ella 4. Tambuson, Kynt

Nathaniel

2. Rocero, Lloyd Andre 5. Ysug, Erwin

3. Suravilla, Kria Faye 6.

EXPERIMENT NO. & TITLE _____________________________________________

Test for Observation

Detection of Inorganic Sulfuric acid White precipitate of barium sulfate forms that indicated
inorganic sulfuric acid is present.

Detection of Ethereal Sulfuric acid White precipitate of barium sulfate forms upon adding
barium chloride that indicated ethereal sulfuric acid is
present.
Calcium Sulfate Crytals Colorless, needle-like, or prism-shaped crystals
indicated that that the calcium sulfate crystals is
present.

2. Chlorides
Observation
Detection of Chlorides Curdy white precipitate

3. Phosphate
Test for Observation
Formation of triple phosphates Absence of white or colorless crystalline precipitates
indicates a negative result for urine infection.

Detection of earthy phosphates Exhibit a yellowish-white precipitate due to the

presence of calcium phosphate.

Detection of alkali phosphates Absence of a visible color change that indicate a

negative result for an elevated urinary pH.

Pathological Constituents
1. Glucose
Test for Observation +/-
Fehling’s test No color change, it remained to be a dark blue precipitate -
after the water bath
Benedict’s test No color change, it remained to be a blue precipitate after the -
water bath
Nylander’s test No color change, it remained to be a light- yellow precipitate -
after the water bath

Is glucose normally present in urine? In what condition can it be detected in urine?

Glucose is not normally present in urine under normal circumstances because the
kidneys typically reabsorb all the filtered glucose back into the bloodstream. However, glucose
can be detected in urine under certain conditions, such as in Diabetes mellitus and renal
glycosuria. In Diabetes mellitus, uncontrolled diabetes, when blood glucose levels are high
(hyperglycemia), the kidneys may not be able to reabsorb all the filtered glucose, leading to
glucose spilling into the urine (glucosuria). While in Renal glycosuria itis a rare condition where
the kidneys are unable to reabsorb glucose properly, leading to glucose being excreted in the
urine even when blood glucose levels are normal.In both cases, the presence of glucose in
 DEPARTMENT OF CHEMISTRY
College of Arts and Sciences
CENTRAL PHILIPPINE UNIVERSITY
ILOILO CITY

CHEMISTRY_5001__ SCORE & TEACHER’S SIGNATURE ______________________

GROUP NO. ___2___ SCHEDULE __________________ DATE _________________

MEMBERS: 1. Eriquez, Micha Ella 4. Tambuson, Kynt

Nathaniel

2. Rocero, Lloyd Andre 5. Ysug, Erwin

3. Suravilla, Kria Faye 6.

EXPERIMENT NO. & TITLE _____________________________________________

urine (glucosuria) can be detected through urinalysis, which is a common test used to screen for
diabetes and monitor blood sugar control in diabetic patients.
2. Albumin
Test for Observation +/-
Heat and Acetic acid test No turbidity observed in the precipitate -
Heller’s test Formation of yellow-orange color at the -
junction
Robert’s test No color formed at the junction -
Is glucose normally present in urine? In what condition can it be detected in urine?
No, glucose is not normally present in urine. However, glucose can be detected in urine
under certain conditions such as diabetes mellitus (both type 1 and type 2), gestational diabetes
during pregnancy, renal glycosuria (a rare inherited condition where the kidneys fail to reabsorb
glucose even with normal blood sugar levels), Fanconi syndrome (a defect in the kidneys that
causes problems absorbing glucose), eating large amounts of carbohydrates at a meal
(alimentary glycosuria), and as a side effect of certain medications like SGLT-2 inhibitors used to
treat diabetes.
3. Blood
Observation +/-
Benzidine The solution is turns from blue color to green +
test

Is glucose normally present in urine? In what condition can it be detected in urine?

No, glucose is not normally present in urine. However, glucose can be detected in urine
under certain conditions such as diabetes mellitus (both type 1 and type 2), gestational diabetes
during pregnancy, renal glycosuria (a rare inherited condition where the kidneys fail to reabsorb
glucose even with normal blood sugar levels), Fanconi syndrome (a defect in the kidneys that
causes problems absorbing glucose), eating large amounts of carbohydrates at a meal
(alimentary glycosuria), and as a side effect of certain medications like SGLT-2 inhibitors used to
treat diabetes.

IV. CONCLUSION
The experiment on urine analysis provided valuable insights into the various
components and properties of urine. The normal color of urine is primarily due to the presence
of urochrome, a pigment resulting from the breakdown of hemoglobin. Factors like concentration
of waste products, hydration levels, and diet can influence urine color. When urine is allowed to
stand exposed to air, it can develop an ammonia-like odor due to the breakdown of urea by
bacteria. Urine may also darken in color due to oxidation, and substances like urates,
phosphates, and crystals may precipitate out, forming sediments. The normal pH of freshly
voided urine typically ranges from 4.6 to 8.0, commonly falling between 5.0 and 6.0. This acidity
or alkalinity is due to the presence of acidic substances like uric acid and alkaline substances
like bicarbonate ions. Allowing urine to stand without a preservative can lead to microbial
contamination, changes in pH, and the formation of sediments, which can affect the reliability of
 DEPARTMENT OF CHEMISTRY
College of Arts and Sciences
CENTRAL PHILIPPINE UNIVERSITY
ILOILO CITY

CHEMISTRY_5001__ SCORE & TEACHER’S SIGNATURE ______________________

GROUP NO. ___2___ SCHEDULE __________________ DATE _________________

MEMBERS: 1. Eriquez, Micha Ella 4. Tambuson, Kynt

Nathaniel

2. Rocero, Lloyd Andre 5. Ysug, Erwin

3. Suravilla, Kria Faye 6.

EXPERIMENT NO. & TITLE _____________________________________________

urine testing and analysis. Uric acid is a normal waste product in urine, but abnormal levels may
indicate health conditions like gout or kidney stones. Indican is normally present in urine at low
levels, and elevated levels may suggest digestive issues or bacterial overgrowth. Hippuric acid
is a normal constituent of urine, formed from the breakdown of certain compounds. The
pigments normally present in urine include urobilin, which can be detected through the
Ammoniacal Zinc Chloride test. The purine bases normally present in urine are adenine and
guanine. Glucose is not normally present in urine, but it can be detected in conditions like
diabetes mellitus and renal glycosuria. Overall, this experiment provided a comprehensive
understanding of the various components and properties of urine, which is crucial for
interpreting urinalysis results and diagnosing potential health conditions if possible.

V. REFERENCE