Biochemistry Laboratory 5049 Date: 07/13/2022

Narrative Report
Narrative Report
Topic Outline

Laboratory Activity 5: Lipids

Laboratory Activity 6: Analysis and Denaturation of

Proteins

Laboratory Activity 7: Isolation of DNA from

fruits/Vegetables and Human DNA

Laboratory Activity 8: Enzymes and factors thatv

affect enzyme activity

Laboratory Activity 9: Optimum ph and Temperature

of hydrolitic enzymes

Laboratory Activity 10: Analysis of Urine

Labvoratory Activity 11: Analysis of Feces

Laboratory Activity 12: Analysis of Blood GROUP MEMBERS

Laboratory Activity 13: Analysis of Saliva Hazel Mae Alindada

Trisha Mae Infante

Rachelyne Matulin

Shaynelle Garnette Paranis

INTRODUCTION Blessie Anne Prado

Sharelle Rimas
Biochemistry focuses on understanding the chemical
basis which allows biological molecules to give rise
INSTRUCTOR
to the processes that occur within living cells and
between cells, in turn relating greatly to the Sir. Albert C. Cabullos
understanding of tissues and organs, as well as
organism structure and function.
 Laboratory Activity 1
Lipids
Water does not easily dissolve lipids, but organic solvents like benzene or chloroform do. Their roles include
serving as cellular membrane components, metabolic fuel, and energy stores. Fatty acids, triglycerides, sphingolipids, and
steroids are different types of lipids.

Soaps are the alkali metal salts of fatty acids that typically include 10 to 18 carbon atoms. They have a long,
nonpolar hydrocarbon chain that is oil-soluble at one end and a carboxylate ring that is water soluble at the other. Because
of this, soap functions well as an emulsifying and wetting agent. Soap is an effective cleaning agent since it can suspend
materials that won't dissolve in water. But when combined with metal ions like Co and Mg ions, soaps create insoluble
salts. Hard water contains these lons, which produce precipitates that stick to sinks and even clothing. The issue of "scum"
creation with soap was resolved with the invention of synthetic detergents.

By including iodine in CC14, it is possible to assess the level of ipid unsaturation. Through double bonds, the free
12 binds to the carbons. The decolorization of the additional lodine is a sign of unsaturation.

We conducted three procedures as part of the laboratory activity for lipids on June 23 of the year 2022. In order to
test for unsaturation, we first combined 5 drops of each of the following oils in separate test tubes: olive oil, oleic acid,
stearic acid, peanut oil, canola oil, com oil, linseed oil, cottonseed oil, and coconut oil. We then added CCI4 drop by drop,
shaking the mixture until a reddish brown color persisted. The number of iodine drops taken was then recorded. The
cotton seeds, on the other hand, use the least amount of iodine.

The Lebermann-Burchard Test is the second step. Whereas great care should be taken when performing this test.
A few cholesterol granules were added to 3 mL of onhydrous chloroform in a clean, dry test tube. 10-20 drops of acetic
anhydride were added, followed by 2 drops of conc. carefully. After slowly combining H2SO, we saw the creation of a
lilac tint that progressively evolved into blue and, eventually, an emerald green color.

Finally, we made a solution of 1 g of powdered detergent in 50 mL of distilled water, checked the pH of the
detergent solution and soap solution by adding phenolphthalein, and then poured 10 mL of the soap solution into two test
tubes with labels before adding 2 mL of the 0.5 percent. Now comes CaCl was placed in test tube 1, 0.5 percent MgCh
was placed in test tube 2, and the process was repeated using detergent solution in place of soap solution. After that, we
added 1 mL of cottonseed oil, 10 mL of warm water, 1 mL of soap solution, and shook the test tube once more. Replace
the soap with detergent and repeat this process. In CaCl2 solution on soap, it has a cloudy precipitate while in detergent, it
was immiscible, for the MgCl2 solution, it has clear precipitate in soap and also immiscible in detergent. Lastly, for the
emulsification, there is a mixed and has a cloudy formation in soap and on the otherhand, there is dispersed in detergent

Laboratory Activity 7
Analysis and Denaturation of Proteins
On the 4th of July, 2022, we did an activity regarding the analysis and denaturation of proteins. Denaturation is
the process of unfolding the complex secondary, tertiary, and quaternary structures of proteins. Proteins function as
biological catalysts or enzymes that transport oxygen and hormones in our body.

The items and materials listed below are also used and provided for our laboratory experiment. Throughout the
whole experiment, we perform different procedures with different tests in them.

In Protein Denaturation, the following materials are utilized in our experiment: 2 fresh eggs, 10-20 mL of
calamansi juice, baking soda, 100 mL distilled water, 70% ethyl alcohol, a dropper, and (optional) inorganic fertilizer). In
order to make an albumin solution, we separate the yolk and the eggwhites of two eggs. Then, in a 100 mL of distilled
water, we put the eggs and stir them until they mix. Then we let it stand at room temperature. In the procedure of
coagulation by heat, we get 2 test tubes and label them 1 & 2. In both test tubes we placed a small amount of albumin
solution. In TT1 and TT2 we add 3 mL of distilled water and heat it in the boiling water bath for 10 minutes with constant
stirring. We allow it to cool, and after that we observe. Upon observation, we notice that TT1 and TT2 are both dim white,
but TT1 has more precipitate than TT2. After that, we move forward in the Inorganic Acids test. We get another glass
container and label it. After that, we add 5 mL of albumin solution, then put 2 mL of calamansi juice, and then we observe
it. Upon observation, we noticed that when the solution is heated, a white coagulant or coagulum is obtained because
albumin is denatured by heat. After that, we moved forward in the alcohol test. We add 5 mL of albumin solution, then
add 3 mL of 70% ethyl alcohol and then we observe it. Upon observation, we noticed that it formed a three-layer of liquid
inside the test tube. The first layer is clear with a few precipitates, the second layer is cloudy with a few brown
precipitates, and the third layer is dirty white with a few brown precipitates in its CH bottom. After that, we move forward
in the salt test. We add 5 mL of albumin solution, then add a pinch of salt to the solution, and then we observe it. Upon
observation, we noticed that the color of the liquid was dirty white with a few brown precipitates at the bottom.

Laboratory Activity 8
Isolation of DNA from fruits/Vegetables and Human DNA

On the 4th of July, 2022, we did an activity regarding the isolation of DNA from
fruits/vegetables as well as us, humans. The main objectives of performing this activity are to isolate
the DNA as well as to discover its fundamental properties and the methods required to isolate it.

The following items were utilized in our experiment: any fruit or vegetable ( in our case, we
used calamansi), detergent, salt, 70% isopropyl alcohol, mortar and pestle or a blender, beakers,
graduated cylinder, glass rod, and lastly, cheesecloth/gauze.

In terms of DNA isolation from fruits or vegetables, we begin by cutting our fruit until 30-50g is
obtained. After that, we transferred the fruit to a mortar and pestle. Then we prepared a precipitating
solution by transferring 30 of warm water into a container, and then we added 1 teaspoon of detergent
and 1/2 teaspoon of salt. We mixed it well until it was dissolved. After that, we transferred the mushed
fruit to a beaker containing the solution and stirred it for 5 minutes. Then we filtered the mixture by
using a cheesecloth into a new beaker and tried to obtain as much liquid as possible. After that, we
added cold isopropyl alcohol in the same amount as the mixture in a test tube, and finally, we tilted our
test tube and slowly poured our isopropyl alcohol into the tube until it formed a layer on top of the
mixture.

As for the second procedure, which is the isolation of a person's DNA, we first added a
tablespoon of salt to 2 cups of water and then used it to gargle for about 1 minute. After gargling, we
spit it in a glass cup and added 2 drops of dish soap in the gargle mixture. After doing so, we added
rubbing alcohol in the same volume as the mixture, waited for 3 minutes, and looked for the DNA
floating in the solution.

With all the tests being accomplished, we were able to come to the conclusion that all living
things have DNA within their cells as it contains all of the necessary building blocks to build and
maintain an organism. The role of the tenderizer in the second method was to dissolve the peptic bond
that connects the amino acids in protein, which releases the links holding those proteins together, which
then makes DNA more accessible. The role of salt in the DNA solution is to neutralize the negatively
charged phosphate in DNA, and the hydration shell of water molecules surrounding the phosphate is
then removed by isopropanol and ethanol. With that being said, the neutralizing of charge and the
elimination of water that causes the precipitate from the solution have the combined effect of allowing
DNA molecules to bind together.

Laboratory Activity 11
Analysis of Feces

The feces include the residue remaining in the intestine after the digestion and absorption of the
food together with products of intestinal secretion, epithelial and bacterial growth and decomposition.
Nowadays, the clinical importance of the study of the stool lies in the microscopic examination
especially in the presence of the parasites and their ova.

The study of the feces includes a macroscopic, microscopic and chemical examination. The
quantity, color, odor, form and consistency, mucus concretion, animal parasites, and curds examination
are included in the microscopic study.
 The chemical examination is a study of the reaction, fermentation, blood and bile pigments in
the feces. Occult blood is one which could not be seen even with the microscopic but only by chemical
means.

The objectives of this experiment is to observe and record the general or physiological
characteristics of feces, to test for the presence of occult blood in the stool, and to test for the presence
of bile pigments in the stool.

The sample used has a yellowish brown color, normal foul odor, acidic pH, smooth and mushy
consistency, and small seed-like particles. Usually, these are the characteristics of a normal stool. The
brownish color of the stool is due to the bile and bilirubin present. The foul smell of a stool is due to
the food that the person eats and the bacteria present in the digestive tract. The slightly acidic pH of a
stool is caused by colonic fermentation of normal amounts of carbohydrate sugars and production of
fatty acids. The sample stool has a smooth, mushy consistency since the baby is just 11 months old and
mostly milk-fed. The little seed-like particles are undigested milk fat. On the detection of bile pigment
derivative (hydrobilirubin), the result and observation in Schmidt’s is upon the addition of saturated
mercuric chloride solution in the fecal matter, they suddenly reacted and produced a pink solution.
After 24 hours, the color of the sample intensified and darkened. While on the Schlesinger’s

There is a very little greenish fluorescence found in the upper part of the solution when exposed
to sunlight. The greenish fluorescence was not visible with the light from the light bulb nor flashlight.

Laboratory Activity 12
Analysis of Blood

The body's most active transport system is blood, a form of connective tissue with a liquid
matrix. The body's most physiologically active tissue is blood plasma. The plasma's proteins are
diverse and complicated, performing a variety of tasks. The three most significant nes are fibrinogen,
globulin, and albumin.

For General test for Blood10 mL of recently taken blood should be put in a heparinized conta iner.
Shake, then put through the following evaluations. Next is Hemin Test Add 1 drop of 1 percent NaCl in
glacial acetic acid after 1 drop of heparinized blood on a glass slide. The mixture should evaporate over
a water bath. Cool and check with a microscope. Draw and label what you have observed. Benzidine
Test, One drop of blood should be diluted with 100 mL of distilled water. Add 6 mL of a 1% benzidine
solution and 2 mL of a 3% H2O2 solution to 2 mL of the diluted blood. Watch for the development of
green or blue color. The outcome is described below. What does the development of blue color mean?
Note the reaction that occurred. Test for Blood Constituents, In a tiny beaker, combine 2.5 mL of
recently drawn blood or defibrinated blood with 25 mL of distilled water. Add 1–2 drops of 10% acetic
acid once the water has boiled, and then keep heating for a few more minutes. When the solution is
heated, filter it. Use the filtrate for the subsequent tests and set aside the residue for the iron test. Test
for Chlorides, We Add 2 to 3 drops of a 5 percent AgNO3 solution after 0.1 M HNO3 has been used to
acidify 1 mL of filtrate. Chlorides are present when a white precipitate forms. Test for the Presence of
Phosphates, Add 4 mL of the 5 percent ammonium molybdate solution after acidifying 2 mL of the
filtrate with 0.1M HNO3. The appearance of yellow precipitate indicates a successful outcome.
Benedict's Test, In a test tube, add 4 drops of Benedict's reagent after adding 2 mL of the filtrate.
Observe the creation of a brick-red precipitate as the mixture is heated over a water bath. Test for Iron,
To completely burn out all organic material, heat the leftovers in an evaporating dish. 2 to 3 mL of a
10% HCl solution should be added after cooling. The dissolved substance has to have 3 to 4 drops of a
5% KCNS solution added to it. Should be observed is the resulting color.

For the Results and Discussion, Chlorides is no white precipitate, which means there is no chloride
present. For the Phosphates yellow precipitate is present, which means the test was successfull. And
lastly, Benedict's Test is brownish green in color and has no precipitate Iron- The solution's color
matches the burn out, which changes from translucent to light yellow.

Laboratory Activity 13
Analysis of Saliva

Salivary mucin isolation to get 15 mL of saliva, rinse your mouth out with water. To the
mixture, add 3 drops of 1N acetic acid. Add the saliva slowly, stirring the flask after each addition, to
30 mL of acetone in an Erlenmeyer flask. The flask should be sealed, then left alone for 30 minutes.
Use two volumes of 3 mL each of acetone to filter and wash the precipitate that was produced. Allow
the precipitation to completely drain. The filter paper should be taken out of the funnel, stretched out,
and allowed to dry in a watch glass. Test the precipitated mucin for any potential saliva components
( in 2a and 2b)

Carbohydrates In a test tube, put about half of the isolated mucin. Add 1 mL of 3M HCl, then
soak for 30 minutes in a boiling water bath. Benedict's solution (3 mL) is added after cooling and 3
drops of 3M NaOH. For ten minutes, reheat in a bath of boiling water. After 30 minutes of boiling, the
mucin fragmented into tiny pieces. Benedict's solution was added, and after another boil, both the
solution and the particles turned black. Proteins break down the residual mucin in 1 mL of 2.5 M
NaOH before including 3 drops of a 0.05 percent CuSO4 solution. Mix the mucin turned brown, and
the fluid took on a purple hue. Nitrates Blend 1 mL of. 0.05 N H2SO4, two drops of KI solution diluted
at 1 percent, and two drops of starch solution. Spit 5 mL into the mix. The saliva was dissolved in the
acetone mixture as the mixture turned yellow. The bottom is covered in a dark precipitate. Add 5 drops
of freshly made 0.1M FeCl3 solution and 5 drops of 0.1 N HCl to 5 mL of saliva to create thiocyanate.
Saliva remained on top while the O.11M FeCl3 solution and 0.1 N HCl mixture remained at the
bottom. cloudy arrangement. Glucose 15 minutes after a meal, collect a saliva sample. Add 2 mL of
Benedict's solution to 5 mL of saliva, followed by 10 minutes in a hot water bath. Only after being
boiled in a pot of boiling water did the saliva and Benedict's solution mix. It only has an ombre effect.
Mix 5 mL of saliva and 2 mL of the ammonium molybdate solution with the inorganic phosphate. After
adding 10 drops of ammonium hydroxide, heat for an additional 5 minutes. Bubbles can be seen in the
top of the saliva, which floats. Ammonium hydroxide and ammonium molybdate settled at the bottom.

Nitric acid, followed by AgNO3 solution, should be added to 5 mL of saliva before adding the 5
drops of chloride. There is something resembling a foggy structure, and at the bottom is a yellow liquid
solution.
DOCUMENTATION