Republic of the Philippines

CAVITE STATE UNIVERSITY

Don Severino delas Alas Campus
Indang, Cavite

COLLEGE OF ARTS AND SCIENCES

Department of Physical Science

BIOCHEMISTRY
LABORATORY
EXERCISES AND EXPERIMENTS

Compiled by:
Nancy P. Galit
 TABLE OF CONTENTS

I. Laboratory Safety Rules and Regulations

II. Exercises

Exercise 1: The Cell …………………………………………………...01

Exercise 2: Concentration Problems ………………………………..04
Exercise 3: Preparation of Buffers…………….…………………..….05

III. Experiments

Experiment No. 1- Buffers in Action ………………………….……..09

Experiment No. 2 - Water and its Properties……….……….……...11

Experiment No. 3 - Protein Analysis…………….….……….….……13

Experiment No. 4 - Denaturation of Proteins………………..………16

Experiment No. 5 - Carbohydrates…….……….…….……………...18

Experiment No. 6 - Tests for Carbohydrates …………….……..….20

Experiment No. 7 - Properties of Lipids ……………..….………….22

Experiment No. 8 - Lipid Tests……………………..…..……..……..25

Experiment No. 9 - DNA Extraction………………………..……..…27

Experiment No. 10 - Collecting DNA ………..……….……………..29

Experiment No. 11 – Color of Urine …………….……………..…....31

References…………………………..………………………………….32
 Laboratory Safety Rules and Regulations

For Instructors

In this difficult time, our goal is to be able to deliver clear instructions to

our students and to create laboratory experiments they can perform without a teacher
present, while reducing technical glitches. This may seem intimidating especially
at a time when everyone is feeling stressed by our global health crisis. To be able to
modify a previously planned lab so that students can perform it at home. Or, due to limited
resources, the better option might be creating a new lab from scratch by considering the
resources available to your students now. Keeping in mind that students do not always
purchase the required lab materials in a timely manner. Focus on supplies in the home as
much as possible.

For Students:
Home labs are a good learning experience. Students are required to take a
picture of their results with a notecard in the shot that has their name and date on it, to
help verify that they performed the lab.
Students should establish safety precautions at home when they perform the
experiment. They need to follow safety protocols, whether they are using materials they
assembled on their own or other materials and chemicals needed in the experiment.
Students are provided with online safety videos to prepare them before they do
that first lab and apply all the things discussed and explained before the laboratory period.
They should not hesitate to ask questions if in doubt about any procedures, and ask the
teacher for help.
Laboratory gown/apron, safety goggles and gloves must be worn whenever they
perform the experiment.

Exercise No. 1
The Cell
Name: __________________________ Date Submitted: _______________________
Year and Sec: _____________________ Date Performed: _______________________
Group No. ________________________ Score: _______________________________

Objectives:

1. Describe what a cell is.

2. Define and understand the structures and functions of each cell part.
3. Understand the structure and purposes of basic components of cells.

Theory:

The basic unit of structure and function in the human body is the cell. Each of a cell’s part,
or organelles, as well as the entire cell, is organized to perform a specific function. Cells
have the ability to metabolize, grow and reproduce, move and response to stimuli. The
cells of the body differ in shape, size and in specific role in the body. Cells that are similar
in structure and function form tissues, which in turn, construct the various body organs.

I. Using the following list of terms, correctly label all the cell parts indicated by leader lines.
Then select different colors for each structure and use them to color the coding circles and
the corresponding structure in the illustration.

A. Animal Cell
___ Nucleus ___ Golgi Bodies
___ Nucleolus ___ Smooth Endoplasmic reticulum
___ Nuclear membrane ___ Rough Endoplasmic reticulum
___ Plasma Membrane ___ Lysosome
___ Cytoplasm ___ Filamentous Cytoskeleton
___ Mitochondrion (microtubules)

01
II. Anatomy of an Animal Cell
 Complete the following table to fully describe the various cell parts.

Cell Structure Function

B. Plant Cell

___ Nucleus ___ Golgi Bodies

___ Nucleolus ___ Smooth Endoplasmic reticulum
___ Nuclear membrane ___ Rough Endoplasmic reticulum
___Cell Wall ___ Lysosome
___ Chloroplast ___ Vacuole
___ Mitochondrion

02
III. Anatomy of a Plant Cell

Complete the following table to fully describe the various cell parts.

Cell Structure Function

03
Exercise No. 2
Concentration Problems
Name: ___________________________ Date Submitted: _______________________
Year and Sec: _____________________ Date Performed: _______________________
Group No. ________________________ Score: _______________________________

Objective:

Review the students in solving for the concentration of solution in Molarity, Molality,
Normality, percent by mass and percent by volume.

Theory:

The concentration of a solution is the amount of solute present in a given amount of

solvent, or a given amount of solution.

1. A sample of 0.653 g of potassium chloride (KCl) is dissolved in 45.5 g of water. What is

the percent by mass of KCl in the solution?

2. Calculate the molality of a sulfuric acid solution containing 34.2 g of sulfuric acid in
173 g of water. The molar mass of sulfuric acid is 98.09 g/mol.

3. How would you prepare a 1.8% NaBr (w:v) aqueous solution?

4. In a biochemical assay, a chemist needs to add 2.93 g of glucose to a reaction mixture.

Calculate the volume in mL of a 1.35M glucose solution she should use for the addition.

5. The concentrated sulfuric acid we use in the laboratory is 98.0 percent H 2SO4 by mass.
Calculate the molality and molarity of the acid solution. The density of the solution is
1.83g/mL

04
Exercise No. 3
Preparation of Buffers
Name: ____________________________ Date Submitted: ______________________
Year and Sec: ______________________ Date Performed: _____________________
Group No. _________________________ Score: _____________________________

Objectives:

1. To prepare a buffer solution.

2. To study and analyze how well the buffer solution resisted to changes to its’ pH and
compared it to the changes in an unbuffered solution.
3. To study the validity of Henderson-Hasselbach equation to a buffer system.
4. To determine the buffer capacity of a buffer solution

Theory:

A buffer solution is one that is resistant to change in pH when small amounts of

strong acid or base are added. For example, when 0.01 mole of strong acid or base are
added to distilled water, the pH drops to 2 with the acid and rises to 12 with the base. If
the same amount of acid or base is added to an acetic acid – sodium acetate buffer, the
pH may only change a fraction of a unit.
Many buffers are prepared by combining a weak acid and its conjugate (acetic acid
and sodium acetate) or a weak base and its conjugate (ammonia and ammonium
chloride). In general, the pH range in which a buffer solution is effective is +/- one pH unit
on either side of the pKa. The Henderson Hasselback equation:

𝑝𝐻 = 𝑝𝐾𝑎 + log conj𝑢𝑔𝑎𝑡𝑒 𝑏𝑎𝑠𝑒 (𝐴)

𝑤𝑒𝑎𝑘 𝑎𝑐𝑖𝑑 (𝐻𝐴)
There is a limit to the amount of acid or base that can be added to a buffer solution
before one of the components is used up. This limit is called the buffer capacity and is
defined as the moles of acid or base necessary to change the pH of one liter of solution by
one unit.

𝐵𝑢𝑓𝑓𝑒𝑟 𝐶𝑎𝑝𝑎𝑐𝑖𝑡𝑦 = 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑚𝑜𝑙𝑒𝑠 𝑜𝑓 𝑂𝐻− 𝑜𝑟 𝐻3𝑂 +

(𝑝𝐻 𝑐ℎ𝑎𝑛𝑔𝑒)(𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑏𝑢𝑓𝑓𝑒𝑟 𝑖𝑛 𝐿)

Materials:

100and 400 mL glass jars 0.10 M CH3COOH

0.02 M NaOH (soap) NaHCO3
Pipette/dropper NaCO3
pH Meter (in the internet) Bottle

05
Sample Computation:

How would you prepare 10mL of a 0.01M phosphate buffer, pH 7.40, from stock solutions
of 0.10M KH2PO4and 0.25M K2HPO4? pKa of KH2PO4 = 7.20. Prepare 10 mL of a 0.01
M phosphate buffer, pH 7.70, from stock solutions of 0.1 M K2HPO4and 0.2 M KH2PO4.
(pKa for the weak acid = 7.20).

1. Use the Henderson Hasselbalch equation to find the ratio of A

pH = pKa + log [A-] / [HA]

7.40 = 7.20 + log [A-] / [HA]
0.20 = log [A-] / [HA]
1.584893192 = [A-] / [HA]*

*Since [A-] / [HA] = 1.584893192, we can say that [A-] / [HA] = 1.584893192/1. In this
case
[A-] = 1.584893192; [HA] = 1.

2. Calculate the decimal fraction (part/whole) of each buffer component.

A- = 1.584893192 / (1.000 + 1.584893192)
=1.584893192/2.584893192 = 0.61313682
HA = 1.000 / 2.584893192 = 0.38686318

3. Find the molarity (M) of each component in the buffer by simply multiplying the molarity
of the buffer by the decimal fraction of each component.
MA- = 0.01M x 0.61313682 = 0.006131368M
MHA = 0.01M x 0.38686318 = 0.003868632M

4. Calculate the moles of each component in the buffer.

Moles = Molarity x Liters of buffer
moles A- = 0.006131368M x 0.01L = 6.131 x 10-5
moles HA = 0.003868632M x 0.01L = 3.869 x 10-5

5. Calculate the volume of each stock solution required to make the buffer
Liters of stock = moles of the buffer component / Molarity of the stock
LA- = 6.131 x 10-5 moles / 0.25 M = 2.452 x 10-4 L = 245
Liters of stock = moles of the buffer component / Molarity of the stock
LA- = 6.131 x 10-5 moles / 0.25 M = 2.452 x 10-4 L = 245µL
LHA = 3.869 x 10-5 moles / 0.10 M = 3.86 9 x 10-4 L = 387 µL

06
Procedure:

A. Phosphate Buffer

1. Prepare 100 mL of a 0.01 M phosphate buffer, pH 7.70, from stock solutions of

0.1 M K2HPO4and 0.2 M KH2PO4. (pKa for the weak acid = 7.20).

a. Use the Henderson-Hasselbalch equation to calculate the volume of each stock

solution needed.
pH = pKa + log [conjugate base] / [weak acid]
b. Check your calculations with other students. See the instructor if there is
uncertainty.
c. Make the solution and check the pH of a portion of your buffer solution using the
pH meter.

B. Acetate Buffer

2. Prepare 100 mL of 0.01 M acetate buffer,pH 3.80, from stock solutions of 0.1 M
acetic acid and 0.02 M sodium hydroxide. pKa acetic acid = 4.76.

a. Use the Henderson-Hasselbalch equation to calculate the volume of each stock

solution needed.
pH = pKa + log [conjugate base] / [weak acid]
b. Check your calculations with other students. See the instructor if there is uncertainty.
c. Make the solution and check the pH of a portion of your buffer solution using the pH
meter.
d. Calculate the exact volume of the 0.01 M acetate buffer required to make 100 mL of
0.0005 M acetate buffer.
e. Prepare this new buffer using the following equation to aid you in your calculations.
C1V1 = C2V2
d. Check the pH of this new buffer.

C. Bicarbonate Buffer

3. Prepare 100 mL of 0.200 bicarbonate buffer, pH 10 by calculating the mass of sodium

bicarbonate and sodium carbonate.

a. Use the Henderson-Hasselbalch equation to calculate the volume of each stock

solution needed.
pH = pKa + log [conjugate base] / [weak acid]1
b. Check your calculations with other students. See the instructor if there is uncertainty.
c. Make the solution and check the pH of a portion of your buffer solution using the pH
meter.
L = 245μL

07
DATA:

Show your calculation:

1. Preparation of Phosphate Buffer

Calculated Volume
K2HPO4 _________________________________
KH2PO4 _________________________________

Actual pH of the buffer: ____________________

2. Preparation of Acetate Buffer

Calculated Volume
CH3COOH _________________________________
KH2PO4 _________________________________

Actual pH of the buffer: ____________________

3. Preparation of Bicarbonate Buffer

Calculated Volume
CH3COOH
KH2PO4

Actual pH of the buffer: ____________________

Guide Questions:

1. How would you relate the importance of pH to the body fluids?

2. Calculate the pH if 0.30M Acetic Acid, with Ka = 1.8 x 10-5 is added to 0.20M Sodium
Acetate.

3. On the laboratory shelf are 250mM solutions of both Acetic Acid and Sodium
Hydroxide. How would you make a 100 ml solution of 25mM Acetate buffer of pH 5.50
using these stock solution?

08
Experiment No. 1
Buffers in Action
Name: ___________________________ Date Submitted: _______________________
Year and Sec: _____________________ Date Performed: _______________________
Group No. ________________________ Score: _______________________________

Objectives:

1. To identify which of the three buffer samples is the best buffer.

2. To know the role and importance of buffers in the human body.
3. To understand the effect of increasing or decreasing the pH of our blood.

Theory:

A buffer is a solution that can resist pH change upon the addition of an acidic or
basic components. It is able to neutralize small amounts of added acid or base, thus
maintaining the pH of the solution relatively stable. This requires specific and stable pH
ranges. Buffer solutions have a working pH range and capacity which dictate how much
acid/base can be neutralized before pH changes, and the amount by which it will change.

A buffer system in the human body is an interaction between a weak acid-base conjugate
pair that keeps the body at the proper pH. A conjugate acid-base pair is typically
composed of a weak acid and the basic ion formed when that acid loses a hydrogen ion.
Weak acids tend to be organic, such as carbonic acid or acetic acid.

The circulatory system cleans up the acid and carbon dioxide produced by exercise by
taking it into the blood. This could result in a dangerous condition called acidosis, but the
bicarbonate buffer system maintains the blood pH at 7.4. The bicarbonate buffer system
works by donating protons if the substances carried in the blood stream are too basic and
accepting protons if the substances are too acidic. When the blood becomes more acidic
due to exercise, the additional protons from those acids are absorbed by the bicarbonate
in the blood to form carbonic acid. The increase in carbonic acid in the blood stimulates
the lungs to expel more CO2, which eventually causes the acid in the blood to be lowered
back to a normal range.

Materials:
1. dry milk 6. small glasses
2. tang powder 7. medicine droppers
3. Gatorade 8. plastic spoons
4. Lemon juice
5. Baking Soda

09
Procedure:
1. To each 3 glasses, add 20 mL of lemon juice.
2. Then add I tsp of dry milk to the first glass, 1 tsp of tang powder to the second glass
and 1 tsp of Gatorade to the third glass.
3. Stir each glass for 10 seconds to dissolve thoroughly the solid in the lemon juice.
4. Add 1 tsp of baking soda to each glass and observe what happened.
5. Write your observation on the table below:

DATA:

Buffer Concentration Reaction time Result

1. dry milk
2. tang juice
3. gatorade

Guide Questions:
1. Identify the three buffers used in the experiment?

2. Which of the three buffers is the best buffer?

3. What is the concentration of the substance used in the buffer solution?

4. How is the strength of buffer determined or measured in the experiment?

5. What are the systems in our body that act as a buffer? How do they work?

6. What are the acids in our body and how to stabilize them in our body?

7. What is the pH of our blood? What will be the effect if it will increase or decrease than
normal?

Illustration:

Generalization:

10
Experiment No. 2
Water and its Properties
_______________________________________________________________________
Name: ___________________________ Date Submitted: ____________________
Year and Sec: _____________________ Date Performed: ____________________
Group No. ________________________ Score: ____________________________

Objectives:

1. To understand the uses of water as a solvent and as a medium for biochemical

reactions.
2. Describe some of the properties of water, and relate them to hydrogen bonding.
3. Discover the unique properties of water?

Theory

Water is important to all living systems. It serves as a natural solvent for mineral ions and
other substances. It is also the dispersion medium for colloidal cells like protoplasm. It
serves as a medium for most biochemical reactions, and is the most abundant component
of cells. Except for bone tissues and enamel, water constitutes about 70% of the human
body.

Materials:

NaCl Sugar Gelatin

Margarine Ethanol Acetone
citric acid powder NaHCO3 powder
Water

Procedure

1. Water as a universal solvent

a. Put about 0.5 grams of the following substances into six separate test tubes: NaCl,
sugar, gelatin, margarine, ethanol.

b. Add 1 mL water to each test tube and shake vigorously to dissolve the substance. To
substances that did not dissolve, add another 1 mL of water and shake again. Add
another 1 mL to the solids that still not dissolve and shake again.

c. Repeat the solubility test using acetone instead of water.

2. Water as a good medium for biochemical reactions

a. Mix about 0.1 gram of dry, powdered citric acid and sodium bicarbonate (NaHCO 3) in a
dry test tube. Observe if a chemical reaction occurs.
b. Add about 10 mL of water to the mixture and note what happens.
11
DATA

1. Solubility of substances in water and Acetone

Substances Solubility in Water Solubility in Acetone

1. NaCl

2. Sugar

3. Gelatin

4. Margarine

5. Ethanol

Reaction of Citric Acid and Sodium Bicarbonate

_______________________________________________________________________
_______________________________________________________________________
___________________________________________________________________

GUIDE QUESTIONS:

1. Enumerate the different functions of water in living systems.

2. Water as a good medium for biochemical reactions.

3. Properties of Water Solutions

ILLUSTRATION:

GENERALIZATION:

12
Experiment No. 3
Protein Analysis
_______________________________________________________________________
Name: ___________________________ Date Submitted: ____________________
Year and Sec: _____________________ Date Performed: ____________________
Group No. ________________________ Score: ____________________________

Objectives:

1. To detect the presence of protein concentration using various type of protein tests.
2. To determine which color indicates the positive result.
3. To know which reagent is considered a strong oxidizing agent in determining the
presence of protein

THEORY:

The common property of all proteins is that they consist of long chains of α-amino
(alpha amino). Proteins function as biological catalyst or enzymes, transporters of oxygen
and hormones. Twenty different types of amino acids occur naturally in proteins. Proteins
differ from each other according to the type, number and sequence of amino acids that
make up the polypeptide backbone. As a result, they have different molecular structures,
nutritional attributes and physiochemical properties. Proteins are important constituents of
foods for a number of different reasons. They are a major source of energy, as well as
containing essential amino-acids, such as lysine, tryptophan, methionine, leucine,
isoleucine and valine, which are essential to human health, but which the body cannot
synthesize. Proteins are also the major structural components of many natural foods,
often determining their overall texture, e.g., tenderness of meat or fish products. Isolated
proteins are often used in foods as ingredients because of their unique functional
properties, i.e., their ability to provide desirable appearance, texture or stability. Typically,
proteins are used as gelling agents, emulsifiers, foaming agents and thickeners. Many
food proteins are enzymes which are capable of enhancing the rate of certain biochemical
reactions. These reactions can have either a favorable or detrimental effect on the overall
properties of foods.

Materials:
NaOH Hydroxyproline tryptophan
CuSO4 Asparagine phenylalanine
milk water bath water bath
salt Pipette/Droppers plastic spoons
albumin 40% NaOH glycine
2% Ninhydrin in Acetone HNO3 concentrated H2SO4
Proline tyrosine glacial Acetic Acid
 13
Procedure:

BIURET TEST

1. Add 5-10 drops or 2-3 pcs. of each sample in a small glass Jars.
2. Add 10 drops of biuret reagent to each sample.
3. Allow to sit for 5 minutes.
4. Observe the change in color and identify whether it contains protein or not.

NINHYDRIN TEST

1. Take 1 mL test solution in dry test tube and 1 mL distilled water in another tube as a
control.
2. Pour few drops of 2% Ninhydrin in both test tubes.
3. Keep the test tubes in water bath for 5 minutes.

XANTHOPROTEIC TEST

1. Take 1 mL test solution in dry container and 1 mL distilled water in another glass as a
control.
2. Add 1 mL of concentrated HNO3 in all test tubes.
3. Mix well then heat gently the content in test tube in an incline position. Cool the solution
under tap water.
4. Add 2 mL of 40% NaOH to all test tubes.

HOPKINS COLE TEST

1. Mix 1 mL of glysine with 1 mL of glacial acetic acid in a clean test tube.

2. Incline the test tube and slowly add 1 mL of concentrated H2SO4 but do not mix.
3. Two layers should form
4. Let the layers stand and note the color of the interface after 2-3 minutes.
5. repeat using stock solution of tyrosine and tryptophan.

DATA
A. Biuret Test
Sample Color Change Protein (+) (-)
1. Milk
2. Salt solution
3. Albumin

B. Ninhydrin Test
Sample Color Change Protein (+) (-)
1. Proline
2. Hydroxyproline
3. Asparagine
 14
C. XANTHOPROTEIC TEST
Sample Color Change Protein (+) (-)
1. Tyrosine
2. Tryptophan
3. Phenylalanine

D. HOPKIN’S COLE TEST

Sample Color Change Protein (+) (-)
1. Glycine
2. Tyrosine
3. Tryptophan

Guide Questions:
1. What colors indicate the presence of protein in the given sample?

2. Which of the given reagents are considered strong oxidizing agent?

3. What are the roles of proteins inside our body?

ILLUSTRATION:

GENERALIZATION:

https://www.youtube.com/watch?v=y9mSMQLcHrs NaOH and CuSO4

https://www.youtube.com/watch?v=s2Qq2F54wr4 Xanthoproteic Test
https://www.youtube.com/watch?v=b8dHXanlzX0 Ninhydrin Test
https://www.youtube.com/watch?v=aUBQofFZ4xs Hopkin’s Cole Test
 15
Experiment No. 4
Denaturation of Protein
_______________________________________________________________________
Name: ___________________________ Date Submitted: ____________________
Year and Sec: _____________________ Date Performed: ____________________
Group No. ________________________ Score: ____________________________

Objectives:

1. To determine the process of denaturation of proteins.

2. To determine whether all proteins denature at the same temperature.
3. To understand why egg whites and milk are used as antidotes for heavy metal
poisoning.
4. To learn the factors affecting protein denaturation.

Theory

Denaturation is the unfolding of the complex secondary, tertiary and quaternary

structure of proteins. Heat, strong acids and organic solvents can denature proteins. Heat
causes the atoms within the protein molecule to vibrate more rapidly, causing the
hydrogen bonds and hydrophobic interactions to break. Strong acids break salt linkages
by ionizing the carboxylic group, and the alcohol denatures the protein by disrupting the
hydrogen bonds.
Heavy metals like silver, lead and mercury also denature the proteins by combining the
free carboxylate anions of the acidic amino acid with the metal, causing precipitation. This
is the rationale in the use of proteins (egg white) as antidote for heavy metal poisoning.

Materials
70% ethyl alcohol NaCl
Baking soda (NaHCO3) 10-20 mL calamansi juice
2 fresh eggs distilled water
6 Glasses
Hot water

PROCEDURE

Preparation of the Stock solution

1. Separate the yolk from the egg whites.
2. Mix the egg whites with 30 mL of distilled water and stir well.

Coagulation by heating
3. Place 5mL albumin solution in 2 separate small glass jars, label it 1 and 2.
4. In no. 1, add 3 ml distilled water and 10 mL hot water then stir.
5. With jar 2, add 3 mL distilled water, stir and compare the result with jar 1.
 16
Inorganic Acids
6. Place 5 mL of albumin solution in container 3, then add 2 mL of calamansi juice.
7. Observe and record the changes.

Alkaline Reagent
8. Add a pinch of baking soda, (NaHCO3) in jar 4 with 5 mL albumin solution.
9. Observe and record the changes.

Alcohol
10. In jar 5 with 5 mL albumin solution, add 3 mL of 70% ethyl alcohol.
11. Observe and record the changes.

Salts
12. Add a pinch of salt in jar 6 with 5 mL albumin solution.
13. Observe and record the changes.

DATA

SAMPLES Observation Denaturation (+ or -)

Glass 1 With distilled water, o denaturation of protein -
Glass 2 With hot water, denaturation occurs +
Glass 3 With acids, small white particles occur +
Glass 4 With NaHCO3, denaturation occurs +
Glass 5 With alcohol, more whites formed +
Glass 6 With salts, smaller white particles occurs +

Guide Questions

1. Surgical instruments are sterilized by heating them, and alcohol is used as disinfectant
in cleansing the skin prior to an injection. Why are those methods successful in killing
harmful organisms?

2. Explain why egg whites and milk are used as antidotes for heavy metal poisoning.

3. What is protein denaturation?

4. What factors affect protein denaturation?

ILLUSTRATION:

GENERALIZATION:
 17
Experiment No. 5
Carbohydrates
__________________________________________________________________
Name: ___________________________ Date Submitted: ____________________
Year and Sec: _____________________ Date Performed: ____________________
Group No. ________________________ Score: ____________________________

Objectives:

1. To detect the presence of carbohydrate in the different samples using the Iodine test..
2. To determine which color indicates the presence of carbohydrate in a sample.
3. To discover the effect of temperature and role of saliva in a carbohydrate test.

Theory

Carbohydrates are a major source of energy from our diet. It is composed of the elements
C, H, and O. It is also called saccharides, which means “sugars. Carbohydrates are
produced by photosynthesis in plants. Glucose are synthesized in plants from CO2, H2O,
and energy from the sun. Carbohydrates are oxidized in living cells (respiration) to
produce CO2, H2O, and energy. Monosaccharides are the simplest carbohydrates.
Disaccharides consist of two monossacharides. Polysaccharides contain many
monosaccharides.

Materials

Iodine solution glass jars or vials stirring rod

droppers Starch Sucrose
Brown Sugar Rice w/ Saliva (boiled) Bread w/ Saliva (boiled)
Rice w/o Saliva Rice w/ Saliva
Bread w/o Saliva Bread w/ Saliva

PROCEDURE

Iodine test

1. Place 3 drops of each test carbohydrate solution in separate containers/small jars

2. Add 1 drop of the iodine solution to each test carbohydrate solution.
3. Note and record the color of each sample
 18
DATA

Test Sample Observation Result (+/-)

Sucrose

Brown Sugar

Starch

Rice w/o Saliva

Rice w/ Saliva

Rice w/ Saliva (boiled)

Bread w/o Saliva

Bread w/ Saliva

Bread w/ Saliva (boiled)

GUIDE QUESTIONS:

1. What color indicates the presence of carbohydrates in the sample?

2. What has analysis in the saliva done to the carbohydrate in bread and rice? Be specific.

5. Is there a different result between boiled and unboiled saliva? Explain.

Illustration:

Generalization:
 19
Experiment No. 6
Tests for Carbohydrates
__________________________________________________________________
Name: ___________________________ Date Submitted: ____________________
Year and Sec: _____________________ Date Performed: ____________________
Group No. ________________________ Score: ____________________________

Objectives:

1. To perform colour tests for carbohydrates.

2. To examine the reducing and non-reducing properties of several carbohydrates.
3. To hydrolyze acetal groups in sucrose and starch and thereby form products that have
the hemiacetal groups.
4. To dehydrate a carbohydrate with sulfuric acid.

Theory

Carbohydrates are important to a healthy diet. Carbohydrates are the main source of
energy for the body. They are the sugars, starches, and dietary fiber that occur in plant
foods and dairy products. Foods high in carbohydrates include bread, pasta, beans,
potatoes, rice, and cereals. Each gram of carbohydrates provides 4 calories. The body
breaks carbohydrates down into glucose, which is the primary energy source for the brain
and muscles. Carbohydrates are one of three macronutrients, which are nutrients that the
body needs in larger amounts. The chemical structures of carbohydrates contain carbon,
hydrogen, and oxygen atoms.

Materials

glass jars or vials wash bottle Bial’s Reagent

droppers water bath starch
test tube holder/clamp Saliwanoff’s Reagent glucose
Graduated Cylinder conc. H2SO4 Xylose
Alcohol lamp Molisch reagent Lactose
match 15% FeCl3

PROCEDURE

MOLISCH TEST

1. Label each test tube. Distilled water is the control tube.

2. Add 4 mL sample in each test tube.
3. Add 2 drops of Molisch reagent in each test tube. Start off with the control tube.
4. Incline the test tube and add cautiously 5 mL of H 2SO4.
5. Observe the solution.
 20
BIAL’S TEST
1. Prepare another set of test tubes, place 1 mL of the sample in each test tube.
2. Add 3 mL of the Bial’s orcinol reagent.
3. Heat until the solution begins to boil.
3. Add 1mL of 15% FeCl3.
4. Observe the solution.

SALIWANOFF’S TEST
1. Prepare another set of test tube. Place 1 mL each of the sample.
2. Add 4 mL of Saliwanoff’s reagent in each test tube.
3. Place in a water bath for 1 minute.
4. Continue heating and observe the color change at a 1 minute interval.

DATA

Sample Molisch Reagent Bial’s Reagent Saliwanoff’s Reagent

1. Control
2. Glucose
3. Starch
4. Xylose
5. Lactose

GUIDE QUESTIONS:

1. What are carbohydrates?

2. What are the important things to remember in dealing with concentrated acids?

3. What are reducing and non-reducing sugars?

ILLUSTRATION:

GENERALIZATION:

https://www.youtube.com/watch?v=CLl9tAoLkjw
 21
Experiment No. 7
Properties of Lipids
__________________________________________________________________
Name: ___________________________ Date Submitted: ____________________
Year and Sec: _____________________ Date Performed: ____________________
Group No. ________________________ Score: ____________________________

Objectives:

1. To detect the presence of lipids and related substances in different cooking oils.
2. To observe the solubility of lipids in polar and non-polar solvents.
3. To compare saturated and unsaturated fatty acids in their chemical reaction with Iodine.
4. To identify which is a better emulsifying agent between soap or detergent.

Theory

Lipids are poorly soluble in water but they dissolve in organic solvents such as benzene
and chloroform. Their functions are to act as metabolic fuel, as stored forms of energy,
and as components of cellular membranes. Lipids are classified into fatty acids,
glycerides, sphingolipids, and steroids. Fatty acids are the smallest unit of lipids. They are
further divided into saturated and unsaturated. Saturated or single bonded fatty acids are
usually solid at room temperature. Unsaturated fatty acids contain double bonds and are
liquid at room temperature. Fatty acids are long chained carboxylic acids which, when
ionized, cause the formation of H+ and RCOO- anion. Fatty acids react with NaOH
producing soaps.

Materials

Palm Oil Vegetable Oil Coconut Oil Acetone

Sesame Oil Canola Oil Pork Oil Lard
Margarine Butter Dropper Soap
Iodine solution Olive Oil Detergent Pipette
Glass container/jar Weighing scale

PROCEDURE

1. The Solubility of Fats.

a. The following procedure is to be repeated for each of the samples of lipids at
your table.
b. Place 5 drops of and oil or a small sample of your lipid into each of three
separate test tubes.
c. To the first tube add 5 ml. of water, to the second 5 ml. of acetone and to the
third add 5 ml. of water to which a pinch of soap has been added.
d. Shake each tube well and allow to stand for a few minutes.
e. Observe whether solution or emulsification has occurred. Instead of using
soap add a small quantity of protein to the lipid and water mixture
 22
2. A Test for the Degree of Saturation of Fatty Acids.
a. Add 5 ml. of your liquid lipids into separate test tubes and a control tube with
5 ml. of water.
b. To each tube slowly and carefully add the halogen water dropwise, shaking
the tube after each addition. The halogen solution should be added just until
it fails to be decolorized.
c. Record the number of drops needed to bring about full decolorization.
d. Compare your results with those of your neighbors at other tables and order
your lipids in terms of increasing degrees of unsaturation.

DATA

Solubility of Lipids

Sample Lipid Water Acetone Soap

1. Canola Oil
2. Coconut Oil
3. Butter
4. Margarine
5. Palm Oil
6. Pork Oil
7. Sesame Oil
8. Vegetable Oil
9. Olive Oil

Test for the Degree of Saturation

Saturated or
Sample Lipid Number of Drops Color after adding I2 Unsaturated
1. Canola Oil
2. Coconut Oil
3. Butter
4. Margarine
5. Palm Oil
6. Pork Oil
7. Sesame Oil
8. Vegetable Oil
9. Olive Oil
 23
GUIDE QUESTIONS:

1. Note the difference fats tested for unsaturation. List them from the most unsaturated to
the least unsaturated.

2. From your observations, which is the better emulsifying agent, soap or detergent?

ILLUSTRATION:

GENERALIZATION:
 24
Experiment No. 8
Lipid Tests
__________________________________________________________________
Name: ___________________________ Date Submitted: ____________________
Year and Sec: _____________________ Date Performed: ____________________
Group No. ________________________ Score: ____________________________

Objectives:

1. To examine the chemical properties of lipids.

2. To classify the different lipids based on chemical reactions.
3. To detect the presence of lipids and related substances in a given sample.
4. To differentiate fats from oils.

Theory

These organic compounds are nonpolar molecules, which are soluble only in nonpolar
solvents and insoluble in water because water is a polar molecule. In the human body,
these molecules can be synthesized in the liver and are found in oil, butter, whole milk,
cheese, fried foods and also in some red meats. Lipids are a family of organic
compounds, composed of fats and oils. These molecules yield high energy and are
responsible for different functions within the human body. Lipids are the polymers of fatty
acids that contain a long, non-polar hydrocarbon chain with a small polar region containing
oxygen. Lipids can be classified into two main classes: Saponifiable and Nonsaponifiable
lipids. A saponifiable lipid comprises one or more ester groups, enabling it to undergo
hydrolysis in the presence of a base, acid, or enzymes, including waxes,
triglycerides, sphingolipids and phospholipids whereas a nonsaponifiable lipid cannot be
disintegrated into smaller molecules through hydrolysis, which include cholesterol,
prostaglandins, etc.

Materials:
Spam Coke Coconut Oil Milk
Gelatin Potato Honey Starch
Butter glass jars stirring rod

PROCEDURE

1. Prepare 9 pieces of small paper bags cut into small squares.

2. Smear each sample onto each piece of paper bag.
3. Let it stand for 10-15 minutes until it dries up.
4. If the paper becomes translucent or clear (almost see through) then the sample is a
lipid.
 25
DATA

Sample Observation + or –

1. Butter
2. Coconut Oil
3. Spam
4. Coke
5. Potato
6. Honey
7. Starch
8. Milk
9. Gelatin

GUIDE QUESTIONS

1. Describe lipids.

2. What are the major functions of lipids?

3. What is the difference between fats and oils?

4. Explain the difference between saturated and unsaturated fats.

ILLUSTRATION:

GENERALIZATION:

https://www.youtube.com/watch?v=tvl5uciEGU0&t=65s
 26
Experiment No. 9
DNA Extraction
__________________________________________________________________
Name: ___________________________ Date Submitted: ____________________
Year and Sec: _____________________ Date Performed: ____________________
Group No. ________________________ Score: ____________________________

Objectives:

1. To learn some basic facts about the DNA molecule.

2. To understand the steps involved in DNA extraction and the chemistry involved in
each step.
3. To observe the white springy substances after mixing with enzyme and alcohol.

Theory

Deoxyribonucleic acid (DNA) is a chemical found in the nucleus of cells that contains the
blueprint for the development and function of living organisms. It’s compared to a set of
blueprints since it contains the instructions on how to build cells. The instructions are
divided into segments along a strand of DNA and are called genes. Genes provide the
code for the production of a protein and control hereditary characteristics such as eye
color or personality behaviors. Proteins determine cell type and function, so a cell knows
whether it is a skin cell, a blood cell, a bone cell, etc, and how to perform its duties.

When you swished the saltwater around in your mouth and scraped your teeth along the
inside of your cheek, you were also collecting cheek cells. The salt helped them clump
together. The degreasing agents in the soap worked to break down the cell membrane to
release the DNA, which is housed inside the cell’s nucleus. Gently mixing the soap and
mouthwash solution ensured you didn’t break up the DNA clumps too much. The rest of
the cheek cells remained in the saltwater-soap solution. The strand of clumped together
DNA would have eventually dissolved in the saltwater, but since it’s not soluble in alcohol,
it precipitates out where the liquid layers meet.
For further study, experiment with different types of liquid soap. Try swishing with plain
water to see how salt affects the experiment. See if using lower-concentration or room-
temperature alcohol changes your results.

Materials

Beaker Test tube with tight fitting lid

Scoop Pipette or Dropper
Safety goggles Gloves
Sodium chloride (salt) Distilled or bottled water
Liquid dish soap or hand soap Stirring rod or wooden skewer (optional)
Small glass vial (optional)
Paper Cup (if alcohol doesn’t come in a dropper bottle)
Isopropyl or ethyl alcohol (at least 70% concentration, higher is better—we used 95%),
chilled in freezer for several hours
 27
PROCEDURE:

1. Create a saline solution in a beaker by adding two lab scoops of salt to approximately
25 ml of distilled water. Stir until the salt is completely dissolved.
2. Pour the saltwater into the paper cup.
3. Without swallowing, drink a mouthful of the solution from the paper cup and swish it
back and forth for at least 30 seconds, occasionally scraping your teeth along the inside
of your cheeks as you do. It’s best to do this with a clean mouth, i.e. not right after
lunch.
4. Spit your mouthwash solution back into the cup. Then bend the cup into a sort of spout
and pour the mouth washed solution into the test tube until it fills about one-half inch of
the bottom of the test tube.
5. Carefully add two drops of the liquid soap.
6. Tilting the test tube approximately 45 degrees, use the pipette (or dropper on the
alcohol bottle) to add 20 drops of the chilled alcohol so it slides down the test tube
without disturbing the solution. Since it’s less dense, the alcohol will sit atop the
mouthwash and soap solution.
7. Tightly put the cap on the test tube and very slowly and gently tilt it upside down then
right side up three times. Do it carefully so as not to make bubbles.
8. Let the test tube sit undisturbed in an upright position for one minute. At this point, you
should begin to see a milky white thread, possibly interspersed with bubbles, appear
between the solution and the alcohol. That’s your DNA! After several minutes, the DNA
should be suspended in the alcohol layer.

Guide Questions:

1. What is the purpose of soap in the experiment?

2. What is the purpose of using ice cold alcohol?

3. Where can the error be in getting only few threads of DNA in the experiment?

Illustration

Generalization:
 28
Experiment No. 10
COLLECTING DNA
Name: ___________________________ Date Submitted: ____________________
Year and Sec: _____________________ Date Performed: ____________________
Group No. ________________________ Score: ____________________________

Objectives:

1. To use some physical and chemical processes to extract a visible mass of DNA from a
fruit tissue
2. To discover which among the fruit samples produces more DNA.
3. To contrast DNA from cheeks and from fruits.

Theory

DNA is present within every living thing, however, cannot usually be seen by the human
eye in its natural state. Using this experiment, you will isolate the DNA so it can be seen.
You will record your data in a journal and/or take photographs documenting your findings.

Materials

Fruit sample (ripe) tap water

15 mL dishwashing soap 5 mL salt
100 mL isopropyl alcohol measuring spoon
Stirrer zip lock bag or any plastic bag
Strainer 3 glass containers

PROCEDURE

1. Prepare the extraction mixture.

2. Measure 50 mL tap water and pour into beaker.
3. Add 15 mL dishwashing soap, 5 mL salt. Stir until it dissolves and set aside.
4. Put the ripe fruit sample inside the ziplock or plastic bag. Use your hands and fingers to
mash the fruits inside the plastic bag.
5. Pour the extraction mixture into the glass. Mix thoroughly.
6. Pour the mixture through the strainer into another glass.
7. Use spoon to press the masked fruit against the strainer forcing more mixture into the
glass.
8. Add 50 mL of the chilled isopropyl alcohol to the solution.
9. Use spoon or wooden stirrer to gently remove the DNA.
 29
GUIDE QUESTIONS:

1. Do you get more DNA from using your chosen fruit? Why do you think this is so?

2. Do you think you can still get results if you skip any step in the procedure? Why?

3. Compare the DNA collected from using your cheeks versus the ripe fruit.

Illustration

Generalization:
 30
Experiment No. 11
Color of Urine
Name: ___________________________ Date Submitted: ____________________
Year and Sec: _____________________ Date Performed: ____________________
Group No. ________________________ Score: ____________________________

Objectives:

1. To gain an understanding of normal and abnormal urine composition.

2. To learn what tests can analyze a person’s urine.
3. To apply knowledge through research works of normal versus abnormal urine to
determine diagnoses on unknown samples and the remedies which can be done to
normalize one’s urine.

Theory

Urine is formed by the kidneys as they function to remove waste products and
foreign materials. It keeps the level of ions in the blood serum at constant value. These
substances include ammonia, urea, proteins from pathogens, uric acid and ions of
hydrogen, sodium potassium, calcium, chloride and sulfate.

Materials

Urine sample
pH meter

PROCEDURE

Physical Properties
1. Place 20 drops of urine in a test tube.
2. Note the odor/ smell, color and clarity of the urine.
3. Dip a piece of pH paper to determine the pH of the urine

Data:

A. Physical Properties

Smell: __________________________________________________________

Color: __________________________________________________________

Clarity: _________________________________________________________

pH: ____________________________________________________________
 31

Color Indication Remedy

1. transparent or
pale yellow
2. Dark Yellow
3. Honey or Amber
4. Pink to Reddish
5. Blue
6. Dark Brown or
Black
B.

GUIDE QUESTIONS

1. What is your overall assessment on the health of the subject based from the results
of the different tests?

2. What indicates that the person is dehydrated?

3. What are the benefits we get from being well hydrated?

4. What is urine’s composition?

 32
REFERENCES
 Lontoc, B. M. et.al. 2000. Biochemistry Laboratory Workbook
 CH104 Paper and TLC.pdf
 PREP of the Buffers at desired pH_ Carroll lab Chap 3(ex
www. xula.edu_Ch emistry documents biolab Caroll…(14-12-03pdf)
 Chang, R. 2010. General Chemistry, McGraw Hill, New York, USA
 https://www.sciencedirect.com/topics/neuroscience/dna-extraction
 https://people.umass.edu/~mcclemen/581Proteins.html
 https://li.wsu.edu/teaching-tool-boxes/options-for-virtual-labs-and-simulations-for-
laboratory-based-courses/
 https://www.medicalnewstoday.com/articles/161547

VIDEOS:

1. Buffer https://www.youtube.com/watch?v=P-R-Cqvb5yo
Demonstration
2. Cool Water https://www.youtube.com/watch?v=auoPKcbLrAA
Experiment
3. Test for Protein, https://www.youtube.com/watch?v=tvl5uciEGU0&t=65s
Starch, and Lipid
Xanthoproteic Test https://www.youtube.com/watch?v=s2Qq2F54wr4
Ninhydrin Test https://www.youtube.com/watch?v=b8dHXanlzX0
Hopkin’s Cole Test https://www.youtube.com/watch?v=_3Act2Js7kU
4. General Test for https://www.youtube.com/watch?v=CLl9tAoLkjw&t=58s
Carbohydrates
5. Test for Lipids https://www.youtube.com/watch?v=81SpohOUHjA
6. What the color of https://www.youtube.com/watch?v=oiCYI5lNflA
your urine says about
your health
33