EXPERIMENT 1

PROTEINS

Protein is the most abundant substance in the cell next to water, comprising 15% of its
over-all mass. Protein is composed of amino acids as its building block. It is linked together with
peptide bonds with a positive charged nitrogen-containing group at one end and a negatively
charged carboxyl-group. Along the chain is a series of different side chains from different amino
acids. Some side chains are neutral, some are acidic, some are basic, and some are classified as
polar or nonpolar. The different tests in this experiment will help you identify the different
types of amino acids present in a protein sample.

OBJECTIVES
• To perform qualitative test for different types of proteins
• To identify proteins based on the different tests performed
• Relate the test results to the chemical structure of each protein or amino acid

PRE-LAB ASSIGNMENT
For the following tests, research on what is the purpose of each test, the reagents
involved in the test, and its positive indicator:
A. Biuret Test
B. Ninhydrin Test
C. Hopkins-Cole Test
D. Sakaguchi Test
E. Xanthoproteic Test

WHAT TO BRING
Aspartame or Equal (5 tabs or 1 sachet)
Glutathione (1 tab)
1 fresh chicken egg

1
MATERIALS
Graduated cylinder Test tube holder
Beaker Test tube brush
Stirring rod Hot plate
Test tubes Water bath
Test tube rack

REAGENTS
Distilled water Hopkins-Cole reagent
10% NaOH Concentrated H2SO4
50% NaOH 0.02% α-naphthol solution
1% CuSO4 solution Sodium hypochlorite
8% Ninhydrin Reagent Concentrated HNO3

PROCEDURE
A. SAMPLE PREPARATION
1. Aspartame or Equal – 1 sachet in 10 ml water
2. Glutathione capsule – Dissolve 1 tab in 5ml water
3. Egg Albumin Solution – Mix 5 ml of egg white in 45ml water

B. BIURET TEST
1. Add 1.0 ml of protein samples in each test tube.
2. To each test tube, add 2 ml of 10% sodium hydroxide solution and mix
thoroughly.
3. Add 2-3 drops of 1% copper sulfate solution to each test tube, mix well and
record your observation.

C. NINHYDRIN TEST
1. Add 2.0 ml of protein samples in each test tube.
2. Add 1ml of 8% ninhydrin solution to each sample. Mix thoroughly.
3. Heat the test tubes in boiling water bath until a color change is observed.

2
D. HOPKINS-COLE TEST
1. Add 2.0 ml of protein samples in each test tube.
2. To each test tube, add 2.0 ml of Hopkins-Cole reagent and mix thoroughly.
3. Tilt the test tube and carefully pour along one side the tube 1.0 ml of
concentrated sulfuric acid.
4. Hold the test tube in an upright position and observe the color of the ring
formed at the interface of the two liquids. (if no ring is visible, gently agitate the
tube to cause a very slight mixing at the surface)
5. Record your observations.

E. SAKAGUCHI TEST (Strictly observe the order of addition of reagents)

1. Add 2.0 ml of protein samples in each test tube.
2. Add to each test tube 1ml of 10% sodium hydroxide and 1.0-ml of 0.02% α-
naphthol solution. Mix thoroughly.
3. After 3 minutes, add 4 drops of sodium hypochlorite.
4. Immediately note the color of the resulting solution because it fades quickly.
Record observations.

F. XANTHOPROTEIC TEST
1. Add 1.0 ml of protein samples in each test tube.
2. Add 1.0 ml of concentrated nitric acid to each sample and mix with a stirring rod.
3. Warm in water bath for 5 minutes. Note the color of precipitate
4. Cool the contents then make it basic by adding 50% sodium hydroxide.
5. Note the changes.

3
 EXPERIMENT 1
DATA SHEET

Name: _________________________________________ Date: ________________

Yr. & Sec.: ______________________ Group No. ____ Rating: _______________

PROTEINS
EXPERIMENT 1

Instruction: Write positive (+) or negative (-) to the test results and color observed.

RESULTS
TESTS
ASPARTAME GLUTATHIONE EGG ALBUMIN

BIURET TEST

NINHYDRIN TEST

HOPKINS-COLE TEST

SAKAGUCHI TEST

XANTHOPROTEIC TEST

4
POST-LAB GUIDE QUESTIONS
1. What is the principle behind each test? Give general chemical equation.
2. What group of amino acids is identified in each test?
3. Account for the difference in color intensity between each sample (if there are
any).
4. Are all the proteins positive for all tests? Why or why not?

5
 EXPERIMENT 2
PROTEIN DENATURATION

Protein denaturation is the modification in conformation of protein accompanied by

disruption and possible destruction of secondary, tertiary, and quaternary structure of protein.
This is brought upon by different types of agents like heat, mechanical disturbance, inorganic,
and organic substances and etc. Loss of solubility in water is the frequent consequence of
protein denaturation. When denatured protein precipitates out, it is called coagulation. This
experiment help you understand the effects brought about by some of these agents that cause
denaturation of proteins.

OBJECTIVES
• To observe the effects of several denaturing reagents on a protein sample
• To differentiate the effect of these denaturing reagents to the protein sample
• To cite practical applications of these denaturing agents.

PRE-LAB ASSIGNMENT
1. Define denaturation.
2. What physical and chemical agents are capable of denaturing proteins? Give the type of
bonds or attractive forces disrupted by these agents.

WHAT TO BRING
1 fresh chicken egg

MATERIALS
Graduated cylinder Test tube holder
Beaker Test tube brush
Stirring rod Hot plate
Test tubes Water bath
Test tube rack

6
REAGENTS
Distilled water 1% BaCl2
Albumin Concentrated H2SO4
95% ethanol Concentrated HNO3
70% ethanol Picric acid
1% AgNO3 Tannic acid
1% CuSO4 Trichloroacetic acid

PROCEDURE
A. Sample Preparation for Proteins
Egg Albumin Solution – Mix 5 ml of egg white in 45ml water
B. EFFECT OF HEAT
1. Add 1.0 ml of egg albumin sample in a test tube.
2. Heat in boiling water for 5 minutes.
3. Compare the results with the standard.

C. EFFECT OF ALCOHOL
1. Place 2.0 ml of albumin solution in 2 separate test tubes and add to each the
following reagents:
a. 5 ml of 95% ethanol
b. 5 ml of 70% ethanol.
2. Shake vigorously.
3. Compare the results.
4. Record your observations as no precipitation, slight precipitation, and heavy
precipitation.

D. EFFECT OF STRONG ACID and BASE

1. Place 2.0 ml of albumin solution in 2 separate test tubes and add to each the
following reagents:
a. 1 ml of 50% NaOH
b. 1 ml of concentrated HNO3
2. Mix thoroughly.
3. Compare the results.

7
E. EFFECT OF HEAVY METALS
1. Place 2.0 ml of albumin solution in 3 separate test tubes and add to each the
following reagents:
a. 1 ml of 1% AgNO3
b. 1 ml of 1% CuSO4
2. Shake all test tubes.
3. Note the color of the precipitates formed. Set aside for 5 minutes.
4. Decant the supernatant liquid and test the solubility of a small portion of the
precipitate in 5.0 ml water.

F. EFFECT OF ALKALOIDAL REAGENTS

1. Place 2.0 ml of albumin solution in 3 separate test tubes and add to each the
following reagents:
a. 1 ml of 1% picric acid solution
b. 1 ml of 1% tannic acid solution
2. Shake all test tubes gently. Note the color of precipitate formed.
3. Compare with the standard.

8
 EXPERIMENT 2
DATA SHEET

Name: ______________________________ Date: ________________

Yr. & Sec.: ______________________ Group No. ____ Rating: _______________

PROTEIN DENATURATION
EXPERIMENT 2

Instructions: Compare the color and precipitations of the results. Write slight precipitation,
moderate precipitation, or heavy precipitation.
DENATURING
OBSERVATIONS
AGENTS

HEAT

95% Ethanol

ALCOHOL
70% Ethanol

NaOH

STRONG
MINERAL
HNO3
ACIDS

9
 AgNO3

HEAVY
METALS CuSO4

Picric acid

ALKALOIDAL
REAGENTS Tannic acid

POST-LAB GUIDE QUESTIONS

1. Explain how each of the agents causes denaturation of the protein.
2. Explain the differences of the results in each group of reagents tested.
3. Give ONE practical application of each agent presented.

10
 EXPERIMENT 3
ENZYMES

Enzymes are substances that act as a catalyst for biochemical reactions by finding a
pathway which has lower activation energy. Each enzyme has an active site where the chemical
reactions occur. The reactant of the said chemical reaction is called substrate. One amazing
property of enzymes is its high specificity; it only acts to a specific substrate or group of
substrates. For example, alcohol dehydrogenase is an enzyme that oxidizes group of alcohols to
aldehyde to carboxylic acid. On the other hand, enzyme like lactase is only specific for the
substrate lactose that breaks it to glucose and galactose. In this experiment, you will experience
the action of some enzymes to its specific substrate.

OBJECTIVES
• Demonstrate the catalytic action of enzymes through different organic specimen
• Recognize the specificity of enzymes to its substrate
• Identify the products of each reaction through different color tests
• Describe the role of enzyme in digestive processes

PRE-LAB ASSIGNMENT
For the following group of enzymes, research on the role or function of each in
biological system and give one example:
A. Hydrolases
B. Oxidoreductases
C. Lyases

WHAT TO BRING
Potato
Cooked meat or cooked egg white
Saliva
Cheesecloth or gauze pad

11
MATERIALS
Graduated cylinder Test tube brush
Beaker Hot plate
Stirring rod Water bath
Test tubes Thermometer
Test tube rack Blender
Test tube holder Ruler

REAGENTS
Distilled water 0.4% HCl solution
1% starch solution 3% HC2H3O2
Iodine solution (I2 in KI) 3% NaHCO3
1% pepsin solution 3% H2O2

PROCEDURES

A. PEPSIN
1. Place the following into three separate test tubes:
a. 5 ml distilled water
b. 5 ml 1% pepsin solution
c. 4 ml 1% pepsin solution + 1 ml 0.4% HCl solution
2. Add an equal amount of very small piece of cooked meat or small cube of egg
white to each test tube.
3. Warm the test tubes in a water bath (37-40oC) for 1 hour to 1 ½ hour. Shake
occasionally.

B. AMYLASE
1. Put 1.0 ml of 1% starch solution on each two test tubes.
2. To one test tube, add 1.0 ml saliva.
3. Mix both thoroughly and place in a water bath set for 37 oC for 15 minutes.
(Make sure temperature does not raise more than 40oC)
4. Set aside and cool.
5. Add to each test tube 3-5drops of iodine solution. Note the color.

12
C. CATALASE
1. Weigh 10 grams of peeled potato.
2. Mash the potato in a blender.
3. Divide the grounded potato into three test tubes and mark the level of the
potato in the test tube.
4. To the first test tube add 1ml of 3% acetic acid and to the second test tube 1ml
of 3% sodium bicarbonate. Leave the third test tube as is.
5. Add 3ml of 3% hydrogen peroxide to the third test tube and wait for one minute.
6. Measure the height of the bubbles with a ruler.
7. Repeat procedures 5 and 6 to the other test tubes.

13
 EXPERIMENT 3
DATA SHEET

Name: ______________________________ Date: ________________

Yr. & Sec.: ______________________ Group No. ____ Rating: _______________

ENZYMES
EXPERIMENT 3

OBSERVATIONS AND
RESULTS
ENZYMES
TEST TUBE 1 TEST TUBE 2 TEST TUBE 3

A. PEPSIN

B. AMYLASE N/A

C. CATALASE

POST-LAB GUIDE QUESTIONS

1. What group of enzymes does each enzyme in the experiment belong? What is its
specific function?
2. Explain the mechanism of enzyme action on its substrate presented in the experiment.
Identify the substrate and the products of each digestion performed.
3. Explain the specificity of enzymes on its substrate.

14
 EXPERIMENT 4
FACTORS AFFECTING ENZYME ACTIVITY

Enzymes are known to catalyze a lot of biochemical reactions. Some enzymes can
increase the reaction rate many million times faster than without a catalyst. An example is
OMP, orotidylate decarboxylase, catalyzing 1017 times faster than uncatalyzed reaction. This
means that reaction that would take 78 million years will only take 18 milliseconds with the
enzyme OMP. Enzymes are mostly globular proteins. Some are simple proteins; some are
conjugated proteins. Their action on the substrate can be controlled by adjusting the
temperature, pH, or substrate or enzyme’s concentration. Further, since they are proteins, they
also affected by agents that causes them to undergo denaturation. This experiment shows the
effect of these factors on enzymatic reactions.

OBJECTIVES
• Describe the effect of the factors presented in the experiment on enzyme’s activity.
• Relate the factors to human enzymatic metabolic activities.

PRE-LAB ASSIGNMENT
Give the factors affecting enzyme activity and give the normal conditions for each factor
to make enzyme perform in its optimum activity.

WHAT TO BRING
Saliva and potato

REAGENTS
Distilled water
1% starch solution
Lugol’s solution
3% H2O2
Buffer solution at pH 3
Buffer solution at pH 7
Buffer solution at pH 10

15
MATERIALS
Blender Test tube holder
Graduated cylinders Test tube brush
Beaker Hot plate
Stirring rod Water bath
Test tubes Thermometer
Test tube rack

PROCEDURES
A. EFFECT OF TEMPERATURE
1. Weigh 3 sets of 3 grams peeled potato.
2. Perform the following on each set:
1. Mash the first set of 5g potato in a blender (A1)
2. Place the second 5g in the freezer for 15 minutes (A2)
3. Boil the third 5 gram until soft (A3)
3. Place the A1 in a 10ml graduated cylinder and measure the volume.
4. Add 5ml of 3% hydrogen peroxide to the graduated cylinder and wait for one
minute.
5. Note and record the volume of the bubbles.
6. After 15 minutes, mash A2 in a blender. Repeat procedures 3 to 5.
7. Mash A3 and repeat procedures 3 to 5.

B. EFFECT OF pH
1. Label three clean and dry test tubes as B1, B2 and B3. Into each test tube place
2.0 ml of 1% starch solution.
2. Place in B1 2.0ml of pH 3 buffer; to B2 2ml of pH 7 buffer; and in B3 2.0ml of ph
10 buffer solution.
3. Make sure in this step, the saliva is added simultaneously to all test tubes. To
each test tube, add 1.0 ml saliva.
4. Warm all test tubes in 37oC water bath for 15 minutes.
5. After heating, add 2 drops of Lugol’s solution.
6. Note and record the color. Rate the intensity of color from 1- 3, 1 being
colorless.

16
C. EFFECT OF ENZYME CONCENTRATION
1. Prepare the following weights of peeled potato and mash separately in a
blender:
1. 5 grams (C1)
2. 3 grams (C2)
3. 1 gram (C3)
2. Place the samples in 3 separate 25 ml graduated cylinders and measure the
volume of potato.
3. Add 10ml of 3% hydrogen peroxide to C1 and wait for one minute.
4. Measure the volume of the bubbles.
5. Repeat procedures 3-4 for samples C2 and C3.

Note: Do not perform simultaneously the 3 samples if you can’t record the time
simultaneously.

D. EFFECT OF SUBSTRATE CONCENTRATION

1. Prepare the following weights of peeled potato and mash separately in a
blender:
1. 3 grams (D1)
2. 3 grams (D2)
3. 3 grams (D3)
2. Place the samples in 3 separate graduated cylinders and measure the volume of
potato.
3. To D1, add 10ml of 3% hydrogen peroxide and wait for one minute.
4. Measure the volume of the bubbles.
5. Repeat procedures 3 and 4 but this time use 5 ml of 3% hydrogen peroxide for
D2 and 1 ml to D3.

Note: Do not perform simultaneously the 3 samples if you can’t record the time
simultaneously.

17
 EXPERIMENT 4
DATA SHEET

Name: ______________________________ Date: ________________

Yr. & Sec.: ______________________ Group No. ____ Rating: _______________

FACTORS AFFECTING ENZYME ACTIVITY

EXPERIMENT 4

FACTORS VARIATIONS RESULTS RATE

A1

EFFECT OF
A2
TEMPERATURE

A3

B1

EFFECT OF pH B2

B3

C1

EFFECT OF ENZYME
C2
CONCENTRATION

C3

D1

EFFECT OF SUBSTRATE
D2
CONCENTRATION

D3

18
POST-LAB GUIDE QUESTIONS
1. Explain how the factors presented in this experiment affect enzyme activity.
2. Give the theoretical results of each factor and explain of it coincides with the results of
your experiment.
3. Relate each factor to human metabolic activities.

19
 EXPERIMENT 5
DNA EXTRACTION

Nucleic acid is one of the biomolecules which is compose of monomer units called
nucleotides. A nucleotide is a three-subunit molecule composing of a pentose sugar bonded to
the phosphate group and a nitrogen-heterocyclic base. If the nucleotide is made up of a ribose
sugar, it is named as ribonucleotide. If it is a deoxyribose sugar, then it is named as
deoxyribonucleotide. The polymers are called ribonucleic acid (RNA) and deoxyribonucleic acid
(DNA) respectively. In this experiment you will learn how to extract DNA from plant tissue.

OBJECTIVES
• Extract DNA from plant tissue
• Describe each procedure done during the extraction
• Analyze the extracted DNA through a qualitative test

PRE-LAB ASSIGNMENT
1. Differentiate DNA from RNA
2. What are the factors that stabilize and destabilize the DNA structure?

WHAT TO BRING
2 white onion bulbs or 1 piece banana or 5 pieces strawberries
Coffee filter paper or gauge pad

MATERIALS
Graduated cylinder Test tube holder
Beaker Test tube brush
Stirring rod Hot plate
Blender Water bath
Test tubes Gauze pad or coffee filter
Test tube rack

REAGENTS
Distilled water 1% deoxyribose solution
95% ethanol Diphenylamine solution
1% glucose solution Standard Saline Citrate Solution
1% ribose solution,

20
PROCEDURE
***Make two sets of experiment, one for today’s analysis and one for the next session

A. Preparation of materials and reagents

1. Place 20ml 95% ethanol in the refrigerator to chill it. You will use this in
procedure no. 12.
2. Homogenizing solution. In a 250-ml beaker add 120ml hot distilled water, 1.5
gram NaCl, and 5 ml dishwashing liquid. Mix them together using a clean stirring
rod slowly to avoid foaming of the soap.

B. Extraction of DNA
1. Coarsely chop the onions or banana in small cubes (do not chop too finely) and
blend for 30 seconds.
2. Add 50ml homogenizing solution and blend again.
3. Filter the mixture in a beaker through a coffee filter or gauze. When you filter,
try to keep the foam from getting into the filtrate.
4. Take the 95% ethanol out of the freezer and place slowly about 15ml to the
filtrate using a graduated cylinder.
5. Let the mixture stand for 5-10 minutes undisturbed. Observe for a bubble
formation and DNA will precipitate out of the solution. DNA will appear as white
filamentous material in the ethanol layer.
***For the other set-up, leave it overnight.
6. Spoon the DNA with stirring rod and add SSC solution to dissolve it.

C. Analysis of the extracted DNA: Dische Test

1. Prepare four test tubes with the label: glucose, ribose, deoxyribose, and
extracted DNA.
2. Place 2 ml each of the following solutions that corresponds their labels: 1%
glucose solution, 1% ribose solution, 1% deoxyribose solution, and 1% extracted
DNA solution.
3. Add 3 ml of diphenylamine solution to each test tube and mix well.
4. Heat in water bath for 10 minutes and cool in an ice bath. A clear tube means
absence of DNA or RNA.

21
 EXPERIMENT 5
DATA SHEET

Name: ______________________________ Date: ________________

Yr. & Sec.: ______________________ Group No. ____ Rating: _______________

DNA EXTRACTION
EXPERIMENT 5

Instruction: Write positive (+) or negative (-) to the test results

PROCEDURE OBSERVATIONS RESULTS

EXTRACTION

Glucose

Ribose

ANALYSIS
Deoxyribose

SAMPLE

22
POST-LAB GUIDE QUESTIONS
1. What are the general structures of ribonucleotide and deoxyribonucleotide? Explain the
difference.
2. What are the levels of the structure of either RNA or DNA?
3. Explain the purposes of each step done in the procedure of extracting DNA.
4. What is the purpose of diphenylamine test? What is the chemical reaction involved?
Explain the results of experiment based on the principle involved in this test.

23
 EXPERIMENT 6
LIPIDS

Lipids, one of the major biomolecules in living cells have no common structure unlike
proteins, carbohydrates, and nucleic acids. Lipids are defined as organic substances that are
insoluble (or sparingly soluble) in water but soluble in organic solvents. This physical
characteristic as well as the chemical properties of lipids depends on the presence of carboxyl
groups, number of double bonds, number of hydroxyl groups, and length of carbon chains.
Referring to its structure, lipids can be divided into simple, compound or derived lipids.
Simple lipids are esters of fatty acids and alcohol. Compound lipids are esters of fatty acids and
alcohol that contain other functional group. While derived lipids are lipids that contain
hydrocarbon rings and a long hydrocarbon side chain. The building blocks of lipids differ from
each type but the most common is the fatty acid. This experiment helps you observe and
understand the different properties of lipids.

OBJECTIVES
• Learn how to characterize lipids through different tests
• Identify the type of lipids based on chemical and physical properties

PRE-LAB ASSIGNMENT
1. Give 2-3 examples of the different types of lipids.
2. Draw the structures of the lipid samples in the experiment.
3. What is the positive indicator of Iodine test for unsaturation?
4. What is the positive indicator of Acrolein test?

WHAT TO BRING
Olive oil
Coconut oil
Lecithin
Glycerol

24
MATERIALS
Filter paper Test tube rack
Graduated cylinder Test tube holder
Beaker Test tube brush
Stirring rod Hot plate
Test tubes

REAGENTS
Distilled water 1M NaOH
Dichloromethane Iodine solution (I2 in KI)
Cyclohexane Potassium bisulfate
1M HCl

PROCEDURES
A. TRANSLUSCENT SPOT TEST
1. In a small filter paper, drop a lipid sample. Do this to the four samples
separately.
2. Allow the spots to dry.
3. After drying, hold the filter paper against a light and see any translucent spot.
Record your observation.

B. TEST FOR UNSATURATION

1. Place 3 ml of ether in five separate test tubes.
2. Place another 3 ml of ether in a separate test tube and label it as negative
control.
3. Add the lipid sample in each test tube while add none to the control. Mix
thoroughly.
4. To each test tube add 3 drops of I2 in KI.
5. Shake again and note the changes in color.

25
C. ACROLEIN TEST
1. In a clean crucible, place 0.5 gram of potassium bisulfate and one to two drops of
lipid sample. Then cover.
2. Heat the sample slowly and note the odor produced.
3. Repeat procedures 1 and 2 to three other lipid samples.

D. SOLUBILITY TEST
1. In five different test tubes add 1ml each of water, cyclohexane, and 1M NaOH.
2. Add to each test tube three drops of coconut oil and mix thoroughly. Make sure
you place strictly three drops of sample on each solvent as solubility is affected by
amount.
3. Observe if sample is thoroughly dissolved. See separation of layers to detect
miscibility. Record your observations.
4. Repeat the procedure with the other three lipid samples.

26
 EXPERIMENT 6
DATA SHEET

Name: ______________________________ Date: ________________

Yr. & Sec.: ______________________ Group No. ____ Rating: _______________

LIPIDS
EXPERIMENT 6

Instruction: Write positive (+) or negative (-) to the test results and color observed (if applicable)

RESULTS
SAMPLES
TRANSLUCENT SPOT TEST UNSATURATION TEST ACROLEIN TEST

Olive oil

Coconut oil

Lecithin

Glycerol

SOLUBILITY TEST
Instruction: Write soluble or insoluble to the test results

DILUTE
SAMPLES WATER CYCLOHEXANE
NaOH

Olive oil
Coconut oil
Lecithin
Glycerol

27
POST-LAB GUIDE QUESTIONS
1. What is the purpose of translucent spot test? What is detected by this test? Is it
conclusive? Why?
2. Explain why results in the Test for Unsaturation are positive or negative based on the
structure of the lipid samples.
3. What is the principle behind Acrolein test? Explain why results showed positive or
negative.
4. Explain the solubility test by comparing the structures of the lipid samples and the
solvents.

28
 EXPERIMENT 7
CELL MEMBRANE OSMOSIS

Every cell has cell membrane which separates it from outside environment. It is
composed of double layer of lipids and embedded with proteins. Cell membrane plays an
important role in the transportation of specific substances in and out of the cell. It transports
important nutrients in the cell and also transports toxins out of the cell. This transportation of
different molecules in and out of the cell, follows different pathways depending on the nature
of the molecule. It can be an active transport, passive transport or facilitated transport. This
experiment will help us understand more of the semipermeable property of membranes.

OBJECTIVES
• Observe the travel of water molecules in and out of a sample membrane
• Explain the phenomenon that occurs in the experiment
• Make a graph that shows the relationship between the concentration and change in
length of potato sample

PRE-LAB ASSIGNMENT
1. Give three different functions of cell membrane.
2. Differentiate hypertonic and hypotonic solutions.
3. Define osmosis and give one example.

WHAT TO BRING
3 pcs potato
Cellophane
Rubber bond

MATERIALS
Thistle tube Ruler
Beakers Clamp
Cork borer Iron stand

29
REAGENTS
Brown Sugar Table Salt

PROCEDURES
A. OSMOSIS
1. Cover the thistle tube with cellophane.
2. Dissolve a spoonful of brown sugar in 50 ml of water and set aside.
3. Get a 250ml beaker and set-up the thistle tube upside down into the center of
the beaker and secure with a clamp.
4. Using a dropper slowly add the sugar solution to thistle tube until the “head” of
the thistle tube is full.
5. Fill the beaker with tap water with equal level of the sugar solution.
6. Observe the level of water every 15 minutes to one hour.

B. TONICITY AND DIFFUSION

1. Insert a cork borer to your potato so that you can make at least five potato tubes
per potato.
2. Measure the potato tubes so that all have the same lengths. Record this as your
initial length.
3. Prepare five solutions of salt with masses, 1 g, 2 g, 3g, 4 g and 5 g in 100 ml of
water. Label the beakers 1-5.
4. Place three tubes per beaker and leave for thirty minutes.
5. After 30 minutes, measure the lengths of the potato tubes and record your
lengths in a table.
6. Input your data in excel and make a graph out of it (post lab report).

30
 EXPERIMENT 7
DATA SHEET

Name: ________________________________ Date: ________________

Yr. & Sec.: ______________________ Group No. ____ Rating: _______________

CELL MEMBRANE OSMOSIS

EXPERIMENT 7
A. OSMOSIS
0 MINS 15 min 30 min 45 min 1 hour

0.00 cm

B. TONICITY AND DIFFUSION

Amount of salt Initial Length Final Length Change in Length

1g

2g

3g

4g

5g

31
POST-LAB QUESTIONS
1. Explain the results gathered from part A of the experiment by presenting the principle
behind it.
2. Explain the results gathered from part B of the experiment by presenting the principle
behind it.
3. Explain the graph that you made out from the data and draw a general statement from
it.

32
 EXPERIMENT 8
CARBOHYDRATES

Carbohydrates can be classified into monosaccharide, the monomer unit of

carbohydrates, disaccharide if two monosaccharides are joined, oligosaccharide if few
monosaccharides are joined or polysaccharide if it is already composed of large number of
monosaccharides. Several tests can be used to determine whether a carbohydrate is (a) a
ketone monosaccharide (ketose) or an aldehyde monosaccharide (aldose), (b) a
monosaccharide or a disaccharide, (c) a reducing disaccharide or a non-reducing disaccharide,
or (d) if it is a polysaccharide or not. In this experiment different types of carbohydrates will be
tested through different qualitative tests.

OBJECTIVES
• Perform the different tests for different types of carbohydrates
• Understand the principle behind each test and its purpose
• Quantify the amount of sugar in a urine sample

PRE-LAB ASSIGNMENT
4. What are the different classifications of monosaccharides?
5. In each test in the procedure, determine the following:
a. Purpose of the test
b. Expected results

WHAT TO BRING
URINE SAMPLE - should be taken at most an hour before the analysis

MATERIALS
Graduated cylinder Test tube holder
Beaker Test tube brush
Stirring rod Water bath
Test tubes Hot plate
Test tube rack Microscope

33
REAGENTS
5% glucose Conc. Sodium hydroxide
5% fructose Barfoed’s reagent
5% sucrose Seliwanoff’s reagent
5% lactose Lugol’s Reagent
5% starch 5M nitric acid
5% glycogen Benedict’s Reagent

PROCEDURES
You should perform each test in these samples:
5% glucose 5% starch
5% fructose 5% glycogen
5% sucrose Urine
5% lactose

C. MOORE’S TEST
1. Place 1 ml of each sample in 6 separate test tubes.
2. Add 0.5ml of conc. Sodium hydroxide and mix thoroughly.
3. Boil the test tubes and note color and odor.

D. BARFOED’S TEST
1. Place 1 ml of each sample in 6 separate test tubes.
2. Add in each test tube 1 ml of Barfoed’s reagent.
3. Heat for strictly one minute and observe.

E. SELIWANOFF’S TEST
1. Place 1 ml of each sample in 6 separate test tubes.
2. Add in each test tube 1 ml of Seliwanoff’s reagent.
3. Place in a boiling water for 30 seconds.
4. Observe color changes and record.

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F. IODINE’S TEST
1. Place 1 ml of each sample in 6 separate test tubes.
2. Add in each test tube 1 ml of Lugol’s reagent.
3. Observe color changes and record.

G. BENEDICT’S TEST ON URINE SAMPLE (QUANTITATIVE TEST)

1. In a test tubes place 1ml of urine samples.
2. In another test tube place 1ml of 5% glucose sample as positive indicator.
3. Add 1 ml of Benedict’s reagent to all test tubes and mix thoroughly.
4. Place the test tubes in boiling water and remove immediately if one of the test
tubes already changes its color.
5. Record observations according to the pattern below.

DATA COLOR OF SOLUTION INTERPRETATION

(-) Blue Absent
(+) Green, slight yellow ppt* Present, trace
(++) Green, thick yellow ppt Present, about 1g/100mL
(+++) Yellow, orange ppt Present, about 2g/100mL
(++++) Orange, orange to red ppt Present, more than 2g
*ppt = precipitate

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 EXPERIMENT 8
DATA SHEET

Name: ________________________________ Date: ________________

Yr. & Sec.: ______________________ Group No. ____ Rating: _______________

CARBOHYDRATES
EXPERIMENT 8

Instruction: Write positive (+) or negative (-) to the test results

RESULTS
SAMPLES MOORE’S BARFOED’S SELIWANOFF’S LUGOL’S
TEST TEST TEST TEST

GLUCOSE

FRUCTOSE

SUCROSE

LACTOSE

STARCH

GLYCOGEN

BENEDICT’S TEST Result:

POST-LAB QUESTIONS
4. Explain the results gathered from each test by presenting the principle behind it. What
makes the test positive? Explain why some samples exhibit positive results and why
some have negative results.
5. Explain the result of your urine sample.

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 EXPERIMENT 9
CARBOHYDRATE HYDROLYSIS

Of all the organic carbon on Earth, more than half of those are the carbohydrates starch
and cellulose which are produced by plants. Animals and humans possess enzyme in the body
that breaks down starch into glucose molecules while humans don’t have enzyme cellulase to
break down cellulose into glucose molecules. Animals on the other hand, can’t produce starch,
instead it produces glycogen in order to store excess glucose in the body. These three
carbohydrates (polysaccharides) are polymers of glucose. This means that after complete
hydrolysis, these polymers will produce glucose molecules. This experiment will help us
examine the breakdown of starch and glycogen into glucose molecules.

OBJECTIVES
• Hydrolyze starch and glycogen.
• Examine the progress of hydrolysis through iodine test.
• Correlate the findings to carbohydrate catabolism.

PRE-LAB ASSIGNMENT
1. Draw a representative structure of starch and glycogen.
2. What is the chemical reaction that breaks carbohydrates into glucose molecules?
3. What is the difference between hydrolysis of carbohydrates and glycolysis?

WHAT TO BRING
SALIVA – should be gathered during the experiment

MATERIALS
Spot plate Pasteur Pipets
Beaker Wash bottle

REAGENTS
Lugol’s solution distilled water
5% starch 5% glycogen

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PROCEDURES
A. STARCH
1. Place 2-3 drops of starch solution in 10 different spots in the spot plate.
2. Label the spots from 1-10.
3. To each spot, place 2-3 drops of saliva.
4. To spot 1, add one drop of Lugol’s solution and record color.
5. After one minute, add one drop of Lugol’s solution in spot 2 and record color.
6. Repeat procedure 5 every after one minute until all spots are recorded.

B. GLYCOGEN
1. Place 2-3 drops of starch solution in 10 different spots in the spot plate.
2. Label the spots from 1-10.
3. To each spot, place 2-3 drops of saliva.
4. To spot 1, add one drop of Lugol’s solution and record color.
5. After one minute, add one drop of Lugol’s solution in spot 2 and record color.
6. Repeat procedure 5 every after one minute until all spots are recorded.

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 EXPERIMENT 9
DATA SHEET

Name: ____________________________________ Date: ________________

Yr. & Sec.: ______________________ Group No. ____ Rating: _______________

CARBOHYDRATE HYDROLYSIS
EXPERIMENT 9

SPOT SAMPLES’ RESULTS

#
STARCH GLYCOGEN

10

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POST-LAB GUIDE QUESTIONS
1. When starch is acted upon by salivary amylase, what will happen? What is the end
product of the chemical reaction?
2. When glycogen is acted upon by salivary amylase, what will happen? What is the end
product of the chemical reaction?
3. What spot number did the starch show changes in color? What is the time elapsed in
this stage?
4. What spot number did the glycogen show changes in color? What is the time elapsed in
this stage?
5. Do you think saliva can digest starch? How about glycogen? Justify answer.

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