LABORATORY POLICIES

1. Classroom discipline must be observed at all times. Eating, smoking and playing in the
laboratory room are strictly prohibited. This rule is enforced because you will be handling toxic
chemicals and bacterial cultures. Any student caught violating this rule will be subjected to
disciplinary measures.

2. A student who does not stay in the laboratory room for the entire duration of the class is
considered absent.

3. Protective laboratory gowns or aprons must be worn during the performance of an experiment.

4. Avoid wasteful use of reagents, water and other materials.

5. DO NOT return excess reagents to the reagent bottle. This will cause contamination of the
entire stock. If you poured more reagent than the amount you need, give it to another group
who needs the same reagent.

6. Use appropriate apparatus for measuring chemicals. You have already used the ses apparatus
in Chemistry 1.

7. DO NOT bring reagent bottles to your working areas. The other members of the class may find
it hard to locate them.

8. Record all data and observations initially in pencil, and finalize all answers in ink. ALL FINAL
REPORTS MUST BE WRITTEN IN INK.

9. NO MAKE-UPS ARE ALLOWED. Absence during the performance of an experiment means

zero score for the experiment. Reports submitted by students who were absent during the
performance of the experiment will be returned unchecked with the comment NO CREDIT.

10. DO NOT perform unauthorized experiments.

11. Students who missed practical examinations in the laboratory will not be given make-up
examinations.

WASTE DISPOSAL PROCEDURES

1. Deposit pieces of paper, wood, glass, and other dry wastes in the garbage can.

2. Deposit waste acid solutions into the acid jar. Any other toxic chemical must be disposed of in
the waste can (if solid) or in the sink, followed by large quantities of water (if liquid).

3. Be very careful in pouring and transporting acids and other corrosive chemicals. Acids may be
transported only if the container is properly capped.

4. Precautions and waste disposal procedures are usually included in the write-up of experiments
that generate wastes with some degree of toxicity. Follow them conscientiously. They are
instituted for your safety.
 Operation of the Microscope
Experiment no.1

Theory
The microscope is a mechanical device used in visualizing smaller objects and
organisms which are either alive or dead specimens. It is mostly a biological tool but it is still
being used in organic chemistry and biochemistry to visualize the effects of chemical reagents
in minute parts inside the cells which cannot be viewed by the eye even with the use o
magnifying glass.
The invention of a microscope opens a lot of opportunity in discovering and
understanding many functions and importance in biochemistry and other biochemical
processes. During the early period of 1500 it was thru the accidental discovery of a lens maker
in the name of Anton Van Leeuwenhoek who discovered the first minute microorganism. He
was able to visualize this tiny organism with the use of his special lenses. Around early 1600,
Dutch scientist Zacharias Jannsen compounded the microscope. From then onwards different
types of microscopes have been invented and modified to suit different purposes and functions.
Compound microscope uses light and lenses to enlarge and magnify an image under the stage.
Other microscopes that are highly specific are the Phase-Contrast, Dark field microscope,
Bright field microscope, UVL microscope, Electron microscope which uses the beam of
electrons among others.
The three main parts of the microscope:
1. The Mechanical Parts – the basic parts of the microscope that are used for support
and manipulations/operations of the device.
2. The Illuminating Parts – the portion of the microscope that will supply light to the
image fixed on the slides that are placed on top of the stage.
3. Magnifying Parts – the parts where the lenses are being used to visualize and
enlarge the specimens placed on the slides.
Microscopes should be handles properly and correctly. If microscopes are not handled
well this will result to damage to the parts and breakage of the lenses and even incorrect
visualization of the specimens. Thereby, the wrong results in the experiment will be
observed.

Objectives
1. To be able to familiarize in handling and manipulation of a simple compound
microscope
2. To be able to identify the different parts of the microscope
3. To be able to visualize a given biological sample

Materials
Compound microscope 1 small letter “e” clipping
1 clean glass slide small leaf
1 clean coverslip

Procedure:
1. Acquire a simple compound microscope from the laboratory instructor.

2. Identify the different parts of the microscope and clean the lenses with a lens cleaner
tissue as well as the other parts of the device.
3. Prepare the small letter “e” clipping and place this into the clean glass slide and cover it
using a cover slip. Place this on top of the stage of the microscope and secure using
the stage clip. Be sure that the letter e” is correctly facing you. Visualize his using the
LPO (low power objective) then turn to the HPO (high power objective). Draw and
observe.

4. Obtain a piece of small leaf, cut a small portion of the leaf and place that portion on top
of the glass slide and repeat procedure number 3. Draw and observe the cells of the
leaf.

• Note be careful in changing the lenses from LPO to HPO because abrupt change may
break the glass slide or may break the lenses of the objectives.
 CAGAYAN DE ORO COLLEGE
PHINMA Education Network

Biochemistry Laboratory Final Report

Name: ___________________ Rating: _____________

Course/Year/ Section: ______ Date Performed: ____________
Group no. _______________ Date Submitted: ____________

Experiment no. _____

Title: ___________________________________________________________
________________________________________________________________

Objective: _______________________________________________________
________________________________________________________________
________________________________________________________________

Materials: ______________ ________________

______________ ________________
______________ ________________
______________ ________________

Data Obtained:
Draw and label the different parts of the microscope. Give the function of each part.
Draw the different specimens that have been viewed under the microscope.
Specimens LPO (Low Power Objective) HPO (High Power Objective)
Letter “e”

Leaf specimen

Questions:

1. Why is the letter “e” inverted when viewed under the microscope?
__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________

2. Identify the different types of microscopes and give their uses.

__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________

3. Give the advantages and disadvantages in using the electron microscope.

__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
 The Cellular Component
Experiment no. 2

Theory

Human beings are one of the most diverse species on the planet that have an extensive
supply of cells in their body. In order to understand all chemical and biological processes that
are taking place within the cells one should fully comprehend and understand first the cellular
component and its functions.

Objectives
1. To be able to acquire properly typical human cheek cells.
2. To be able to visualize, identify and draw the parts of typical animal cells.

Materials
Clean glass slides tissue paper
Applicator stick dropper
Drinking water microscope
Alcohol lamp methylene blue (dye)
Tap water

Procedure
1. Clean a piece of glass slide and gently pass it over the flame of the alcohol lamp in
order to remove excess dirt or grease on the slide. The slide should always be held at
the edges to prevent contamination of the surface.

2. Place the glass slide on the table with the flamed portion side-up.

3. In acquiring for the cell sample, get an applicator stick and gently rub the inside of the
cheek wall. Do not rub too hard so as not to induce bleeding or injury to the cheek
wall.

4. Gargle with clean water after acquiring the cells taken from the cheek wall.

5. Apply evenly the tip of the applicator stick that has been rubbed on the surface of the
cheek wall on the clean glass slide. This is now your smear.

6. Lay the slide on the table and allow it to dry in air.

7. Fix the smear on the glass slide by allowing it to pass through over the flame 3 to 5
times.

8. This will cause the cell to adhere to the glass slide properly.
9. Apply the methylene blue over the prepared smear using the medicine dropper. Cover
the entire surface of the smear as much as possible. Wait for 2 minutes.

10. Rinse carefully the stained slide with tap water so as not to dislodge or remove the
stained cell sample from the surface.

11. Blot dry the stained smear with tissue paper.

12. Examine the smear under the microscope.

 CAGAYAN DE ORO COLLEGE
PHINMA Education Network

Biochemistry Laboratory Final Report

Name: ___________________ Rating: _____________

Course/Year/ Section: ______ Date Performed: ____________
Group no. _______________ Date Submitted: ____________

Experiment no. _____

Title: ___________________________________________________________
________________________________________________________________

Objective: _______________________________________________________
________________________________________________________________
________________________________________________________________

Materials: ______________ ________________

______________ ________________
______________ ________________
______________ ________________

Data Obtained:

Schematic Drawing of the Cells:

Prepared Slide Observation of the Cell from the Cheek

Drawing
Draw and label all the parts of a Eukaryotic and a Prokaryotic cell.

Post Laboratory Guide Questions:

1. What is a cell? What is its importance to the human body?

_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
________________

2. How were you able to acquire the cell sample?

_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
________________
 3. Why is the sample being taken from the cheek and not from any other parts of the
body? Explain briefly.
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
________________

Clinical Guide Questions:

1. In terms of genetic cloning, what part of the cell organelle is being modified and being
transferred to from a new clone? Explain briefly.
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
________________

2. What is trisomy 21 syndrome? What part of the cellular organelle is defective?

_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
 Water and Its Properties
Experiment no.3

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

Objective
To determine the properties of water that make it a suitable medium for sustaining life
in biological systems.

Materials
NaCl 0.1M AgNO3
Sugar 10% sucrose solution
Gelatin distilled water
CuSO4 dialysis bag
CCl4 cellophane
Citric acid powder 2 beakers (250 ml)
NaHCO3 powder thistle tube
1% NaCl in starch solution 4 test tubes
test tube rack string or rubber band

Procedures
I. Water is a universal solvent.
a. Put about 0.5 grams of the following substances into 6 separate test tubes: NaCl,
sugar, gelatin, CuSO4. Add 1 ml of 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. To the solids that still did not dissolve, add another ml
of water and shake.
b. Repeat the solubility test using CCl4 instead of water.
c. Describe solubility in both solvents as soluble, slightly soluble and insoluble.
Record observations in the table below.

Substances Solubility in water Solubility in CCl4

NaCl
Sugar
Gelatin
CuSO4

II. Properties of water solutions

a. Dialysis. Obtain a dialysis bag about 20-25 cm long and soak in clean water for
about 10 minutes. Fill with 30 ml of 1% starch-NaCl mixture, tie the bag and rinse
thoroughly with water. Put the bag in a beaker containing deionized water. Adjust
the setup such that the levels of fluids inside and outside the bag are the same.
 After 1 hour, test 1 ml of dialyzate (water in the beaker) with a few drops of 0.1
MAgNO3. Formation of a white precipitate of AgCl confirms the presence of
chloride ions in the dialyzate.

b. Osmosis. Stopper the narrow end of a thistle tube with a finger and fill it with
sugar can juice or a 10% sucrose solution until the solution reaches up to the
base of the tube. Cover the mouth of the thistle tube with a cellophane membrane
and keep it in place with a rubber band. Suspend the thistle tube in a beaker of
water, making sure that the levels of liquid inside and outside the tube are equal.
Observe the difference in solution levels after 30 minutes. Did the sugar solution
migrate inwards or outwards?

CAGAYAN DE ORO COLLEGE

 PHINMA Education Network

Biochemistry Laboratory Final Report

Name: ___________________ Rating: _____________

Course/Year/ Section: ______ Date Performed: ____________
Group no. _______________ Date Submitted: ____________

Experiment no. _____

Title: ___________________________________________________________
________________________________________________________________

Objective: _______________________________________________________
________________________________________________________________
________________________________________________________________

Materials: ______________ ________________

______________ ________________
______________ ________________
______________ ________________

I. Water is a universal solvent.

Substances Solubility in water Solubility in CCl4

NaCl
Sugar
Gelatin
CuSO4

Based on your observations, which solvent dissolves more substances?

__________________________________________________________________________

II. Properties of water solutions

a. Dialysis

Observation:
__________________________________________________________________
__________________________________________________________________
__________________________________________________________________
 b.Osmosis.

Observation:
__________________________________________________________________
__________________________________________________________________
__________________________________________________________________
___

Questions:

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

__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
____________

2. Based on your observations, define dialysis and osmosis. Cite possible applications of
the above procedures.
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
____________

Conclusion:
__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
 Analysis and Denaturation of Proteins
Experiment no.4

Theory
The cell is composed mainly of proteins, and their repeating unit, the amino acid. Protein
function as biological catalysts or enzymes, transporters of oxygen, and hormones.

There are four levels of protein structure: the primary, secondary, tertiary and quaternary
levels. The primary structure is composed of single covalently-bonded amino acids. There are
two types of secondary structure, the α-helix and the β-sheets. The alpha helix structure
contains one strand of amino acid chain that is bonded by intramolecular hydrogen bonds. Two
chains that are linked by hydrogen bonds describe the beta-sheet structure. The tertiary
structure of the protein refers to the combination of either pure helix or of pure beta or a
combination of both. The stabilizing interactions in the tertiary level are (a.) salt linkages,
(b)hydrogen bonds, (c) disulfide linkages, and (d) hydrophobic interactions.

Denaturation of Proteins.

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

structure of proteins. Heat, strong acids and organic solvents (e.g., alcohol) 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 split salt linkages by
ionizing the carboxylic group,
and alc
ohol denatures the protein by disrupting the hydrogen bonds.

Heavy metals ions like silver, lead and mercury also denature 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 (e.g., egg white) as antidote for heavy metal poisoning.

Organic acids, like picric acid and tannic, are used to precipitate the alkaloids giving rise
to the name “alkaloidal reagents”. The anion of the acid will react with the protonated amino
groups of protein, causing disruption of the salt linkages. Leather is manufactured by
coating animal skins with tannic acid, hence the process is known as tanning.

Color Tests of Proteins

Amino acids can be identified by the R-groups attached to the α-carbon that reacts with
specific chemicals. However, since proteins contain various amino acids, one test will not be
enough to identify the amino acids. It would be best to perform numerous tests before
concluding the components of a protein.

1. Biuret Test
This test will give a positive result for compounds that contain 2 or more peptide
linkages. It will give a distinctive purple color which is probably due to the formation of
 a complex by cupric ions with the amino groups called Biuret. Hence, dipeptides and
amino acids like serine and threonine do not give positive results with this test.

2. Ninhydrin Test
When amino acids are sprayed with ninhydrin, it will give a blue to violet colored
result. The presence of amino group including those found in amines and ammonia will
also give the same result except for praline and hydroxyproline that will give a yellow
colored result.

3. Xanthoproteic Test
Nitration of amino acids that contain benzene ring will yield the product
nitrobenzene that will give a yellow to orange coloration. Tryptophan, tyrosine and
phenylalanine will produce nitrobenzene when treated with concentrated nitric acid.
Collagen and gelatin do not give a positive reaction.

4. Millon’s Test
The presence of phenol group in amino acid like tyrosine is nitrated by a solution
of mercuric and mercurous nitrates in concentrated nitric acid. A white precipitate will
start to form, turning brick red on prolonged hating due to the formation of a mercury
complex of nitrophenyl derivatives. Addition of NaNO2 turns the precipitate to darker
pink or red.

5. Hopkin’s Cole Test

The aldehyde present in the reagent will cause the formation of a blue or violet
colored ring due to the formation of a complex between the reagent and the indole ring
of tryptophan. Gelatin and collagen fail to give a positive result with this test indicating
the absence of tryptophan in these proteins.

6. Sakaguchi Test
The combination of alpha-naphtol and sodium hypobromite or hypochlorite will
react with the guanido group giving a red to orange colored complex. This test is used
to identify the presence of arginine. This is an extremely sensitive test and may be used
as a general test for proteins, because all known proteins contain sufficient arginine.

7. Lead Acetate Test (Test for Sulfur)

This test is specific for sulfur-containing amino acids like cysteine and
methionine. The sulfahydryl or disulfide groups are converted to inorganic sulfide, Na2S,
in strongly alkaline solution. This will react further with lead acetate to form a brownish-
black precipitate of lead sulfide(PbS).

Objective
To be able to identify the different amino acids in a protein sample.

Materials
Test tubes Tyrosine
Test tube holder Millon’s reagent
Test tube holder 0.1% NaNO2
 Test tube rack Hopkin’s Cole reagent
dropper conc. H2SO4
pipette 10% NaOH
aspirator 5% lead acetate solution
spatula ammonia water
stirring rod 0.2% urea
Bunsen burner 95% ethanol
Tripod 2% mercuric chloride
Wire gauze 2% lead acetate
Watch glass 2% silver nitrate
Water bath Picric acid
Graduated cylinder Tannic acid
Cork egg albumin (from fresh egg)
5% albumin
5% gelatin
conc. HNO3
conc. NH4OH
5% phenol

Procedures

I. Color Tests for Amino Acids and Proteins

A. Xanthoproteic Test
Place 1 ml each of 5% albumin solution, 5% gelatin solution and tyrosine in separately-
labeled test tubes. Add 5 drops of conc. HNO3 into each tube. Notice the formation of white
precipitate. Mix it thoroughly, heat in a water bath, and note the change in color. Allow it to
cool and add a few drops of conc. NH4OH. Note the intensity of the color.

Xanthoproteic Test Observations

Albumin
Gelatin
Tyrosine

B. Millon’s Test
Place 1 ml each 5%solution of albumin, tyrosine and 5% phenol solution in separately-
labeled test tubes. Add 4 drops of Millon’s reagent into each test tube. Heat in boiling water
bath for 10 minutes, then allow it to cool by placing the tube in running tap water. Then add 4
drops of freshly prepared 0.1% NaNO2 and warm gently. Note any change in the color of the
precipitate.

MIllons’ Test Observations

Albumin
Tyrosine
Phenol
C. Hopkin’s Cole Test
Place 1 ml each of tyrosine, albumin solution and 5% gelatin solution in separately-
labeled test tubes. Add 4 drops of Hopkin’s Cole reagent. Incline the test tube in the test tube
rack. Carefully, allow 2 ml of conc, sulfuric acid to slide down the side of the inclined tube so
that it will form a layer below the protein solution. Take note of the color of the junction of the
two liquids.

Hopkin’s Cole Test Observations

Albumin
Gelatin
Tyrosine

D. Lead Acetate Test

Place 1 ml of 5%albumin solution, 5% gelatin solution in separately labeled test tubes.
Add 5 drops of 10% NaOH, and 3 drops of 5% lead acetate solution into each test tube. Shake
and heat in boiling water bath. Describe the color of the precipitate formed.

Lead Acetate Test Observations

Albumin
Gelatin

E. Biuret Test
Place 2 ml each of 5% albumin and urea in separately-labeled test tubes. Add 1 ml of
10% NaOH solution and about 5 drops of CuSO4 solution. Observe the color that develops.

Biuret Test Observations

5% Albumin
Urea

II. Protein Denaturation

A. Coagulation
1. By Heat
Label 2 test tubs as 1 and 2. To both tubes, place a small amount of egg albumin
solution. To tube 1, add 3 ml distilled water and heat the test tube in a boiling water bath for
10 minutes stirring. Allow it to cool. Add 3 ml distilled water to test tube 2 and stir. Filter both
tubes separately, then test both filtrates using Biuret reagent. Compare the results and record
your result below.

Observations
Test tube 1
Test tube 2

2. Inorganic Acids
 Label two test tubes as 1 and 2, and add 3 ml of 5% albumin into each tube. Test tube
1, add concentrated HCl drop by drop, shaking after each addition. Record the number of drops
of the acid added until a precipitate is formed. Then add an excess amount of HCl and take
note whether it will increase or dissolve the precipitate formed. Repeat the same procedure in
test tube 2, but this time use concentrated H2SO4.

Observations
HCl
H2SO4

3. Alcohol
Place 1 ml each of 5 % solutions of albumin and gelatin in separately-labeled test tubes.
Add 5 ml of 95% ethanol in each test tube, mix and note the formation of precipitate.

Observations
Albumin
Gelatin

B. Precipitation

1. By Heavy Metal Salts

In three test tubes, place 3 ml of egg albumin solution. To test tube 1, add 5 drops of
lead acetate solution. Add excess drop of lead acetate solution and note whether the
precipitate is increased or dissolved. Repeat the procedure by adding 5 drops of silver nitrate
to test tube 2 and 5 drops of mercuric chloride to the third test tube. Describe the results.

Heavy metal salts Observations

Lead acetate
Silver nitrate
Mercuric chloride

2. By Alkaloidal Reagents
Place 3 ml of egg albumin solution in two test tubes. In test tube 1, add 2 ml of tannic
acid solution, and in test tube 2, add 2 ml of picric acid solution. Describe the protein solution
in each of the tubes after the addition of alkaloidal reagents.

Alkaloidal Reagent Observations

Picric acid
Tannic acid

CAGAYAN DE ORO COLLEGE-PHINMA

Biochemistry Laboratory Final Report

Name: ___________________ Rating: _____________
Course/Year/ Section: ______ Date Performed: ____________
Group no. _______________ Date Submitted: ____________

Experiment no. _____

Title: ___________________________________________________________
________________________________________________________________

Objective: _______________________________________________________
________________________________________________________________
________________________________________________________________

Materials: ______________ ________________

______________ ________________
______________ ________________
______________ ________________

I. Color Tests for Amino Acids and Proteins

A. Xanthoproteic Test

Xanthoproteic Test Observations

Albumin
Gelatin
Tyrosine

B. Millon’s Test

MIllons’ Test Observations

Albumin
Tyrosine
Phenol

C. Hopkin’s Cole Test

Hopkin’s Cole Test Observations

Albumin
Gelatin
Tyrosine

D. Lead Acetate Test

Lead Acetate Test Observations

 Albumin
Gelatin

E. Biuret Test

Biuret Test Observations

5% Albumin
Urea

II. Protein Denaturation

A. Coagulation
1.By Heat

Observations
Test tube 1
Test tube 2

2. Inorganic Acids

Observations
HCl
H2SO4

3. Alcohol

Observations
Albumin
Gelatin

B. Precipitation
1. By Heavy Metal Salts

Heavy metal salts Observations

Lead acetate
Silver nitrate
Mercuric chloride

2. By Alkaloidal Reagents

Alkaloidal Reagent Observations

Picric acid
Tannic acid
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 these methods successful in killing
harmful microorganisms?
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
________

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

3. Explain why picric acid and tannic acids are sued in the treatment of burns.
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
________

4. What is the colored precipitate obtained in the sulfur test (lead acetate test)?
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
________

Conclusions
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
 _______________________________________________________________________
________

Enzymes and Factors that Affect Enzyme Activity

Experiment no.5
Theory

Metabolic processes provide energy for the maintenance of life-support systems of the
organism. They consist of degrading chemical compounds of relatively high potential energy
to products with low potential energy. An example of such degradation is the conversion of
glucose to CO2 and H2O. The energy evolved is collected, stored and utilized by the cell for
growth, reproduction, synthesis, repair of cellular materials and other functions which keep life
systems operational. These processes are made possible by the presence of catalytic
substances called enzymes.
As learned from basic Chemistry, catalysts alter the energy activation, Ea, of a chemical
reaction. Increasing the Ea decreases the rate of a reaction (negative catalysis) and
decreasing the Ea increases the rate of reaction (positive catalysis).
Enzymes are complex organic substances produced by an organism to alter the rate of
biochemical reactions that take place in the cell. However, unlike inorganic catalysts, enzymes
show a high degree of specificity, with each enzyme catalyzing only one kind of reaction for
only one kind of substrate. For example, the hydrolysis of sucrose is catalyzed by invertase,
which acts only on sucrose and not other disaccharides and even polysaccharides with facility.
At present, about 2,000 different enzymes have been isolated and characterized. In this
experiment, catalase, an enzyme derived from potatoes and which detoxifies peroxides
produced in the body from the metabolism of amino acids will be isolated and characterized.
All enzymes have a protein component and are affected by a number of physical and
chemical factors that denature proteins, e.g., extremes of pH and temperature, presence of
metal and on-metal ions, presence of alkaloidal reagents, etc. In addition, enzyme and
substrate concentrations also affect enzyme activity. In this experiment, the effect of
temperature and pH on salivary amylase is determined.

Objectives
1. To isolate and determine the properties of catalase.
2. To determine the effect of sulfides on catalase activity.
3. To prove the presence of a protein factor in enzymes.
4. To determine the effect of temperature on enzyme activity.

Materials
1 pc. Small potato with peel 0.1N NaCl
 filter paper 0.05N HCl
6% H2O2 0.05 N NaOH
filter or cotton /gauze blender
saliva filter funnel
starch solution cheesecloth
acidified iodine solution (0.001N) wooden splint
3M NaOH CuSO4 solution
1% starch solution 0.1M NaCl
0.1 N NaCl solution

Procedures
I. A. Preparation of catalase enzyme
Grate a potato, including the peel, into a fine pulp. ( A blender may be used for this.)
Add 50 ml of water and allow the mixture to stand for 10 minutes, swirling occasionally. Strain
through cheesecloth and discard the residue. Filter again through filter paper. (Note: If filtering
through filter paper takes too long, use cotton or gauze.) Use the filtrate for the following test:
Mix 5 ml of the filtrate and 5 ml of 6% H2O2 in a test tube. Immediately insert a glowing
wooden splint. What gas is indicated present?

B. The protein nature of enzymes

To 5 ml of the catalase extract, add 5 ml of 3M NaOH. Mix the two solutions by tapping
the tube against the palm of your hand. Add 5 drops of CuSO4 solution to the mixture. Observe
and record the color.

II. Factors that Affect Enzyme Activity

A. Preparation of enzyme.
Extract 10-15 ml of saliva from a student donor and use it to test for the effect of pH and
temperature.

B. Effect of pH
Label 3 test tubes and mix the substances indicated below.

Tube number 1% starch 0.1M NaCl Acid/base/water Saliva

solution added
1 10 ml 1ml 1 ml 0.05N HCl 2 ml
2 10 ml 1ml 1 ml distilled 2 ml
water
3 10 ml 1ml 1 ml 0.05N 2 ml
NaOH
Mix well by shaking each test tube, and place in a water bath maintained at body
temperature (37°C). Recording the time at 3-minute intervals, test for the presence of starch
using 0.001 N iodine solution. Record the time needed for the blue color of starch with iodine
fail to appear. This means that starch has been completely hydrolyzed to glucose.

Time (minutes) Color of saliva extract with iodine

Test tube #1 Test tube #2 Test tube #3
3
 6
9
12
15
18

C. Effect of Temperature
Label 3 test tubes, add 10 ml of 1% starch suspension and 1 ml 0.1 N NaCl solution.
Immerse Tube #1 in a beaker of water at room temperature, tube #2 in a water bath at 37°C,
and tube #3 in a boiling water bath. Using a pipette, add exactly 2 ml of saliva into each tube.
Mix well and monitor in a manner similar to Part II-B. Note the length of time it takes before
the starch is completely hydrolyzed. Record the time needed before the blue color with iodine
no longer appears.

Time (minutes) Color of saliva extract with iodine

Test tube #1 Test tube #2 Test tube #3
(room temperature) Water at 37°C (boiling water bath)
3
6
9
12
15
18
CAGAYAN DE ORO COLLEGE - PHINMA

Biochemistry Laboratory Final Report

Name: ___________________ Rating: _____________

Course/Year/ Section: ______ Date Performed: ____________
Group no. _______________ Date Submitted: ____________

Experiment no. _____

Title: ___________________________________________________________
________________________________________________________________

Objectives: _______________________________________________________
________________________________________________________________
________________________________________________________________
________________________________________________________________

Materials: ______________ ________________

______________ ________________
______________ ________________
______________ ________________
I. A. Preparation of catalase enzyme
What gas is indicated present?
__________________________________________________________________________

Observation:
________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________

B. The protein nature of enzymes

Observation: color of solution

____________________________________________________

II. Factors that Affect Enzyme Activity

B. Effect of pH

Time (minutes) Color of saliva extract with iodine

Test tube #1 Test tube #2 Test tube #3
3
6
9
12
15
18

In what test tube is there evidence of unhydrolyzed starch after 18 minutes?

__________________________________________________________________________

Observation:
__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________

What is the optimum pH of salivary amylase?

__________________________________________________________________________

Conclusion:
__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________

C. Effect of Temperature

Time (minutes) Color of saliva extract with iodine

 Test tube #1 Test tube #2 Test tube #3
(room temperature) Water at 37°C (boiling water bath)
3
6
9
12
15
18

Questions:

1. Write the equation for the reaction catalyze by catalase.

_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
__________________________________________

2. How does an enzyme alter the rate of chemical reaction?

_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
__________________________________________

3. What is the optimum pH and temperature for salivary amylase?

__________________________________________________________________________
_______
__________________________________________________________________________
_______

4. What effects do a.) increased pH and B.)increased temperature have on the activity of
salivary amylase?
a.) increased pH -
__________________________________________________________________
__________________________________________________________________________
_______
__________________________________________________________________________
_______
__________________________________________________________________________
_______

b.) Increased temperature -

___________________________________________________________
__________________________________________________________________________
_______
__________________________________________________________________________
_______
__________________________________________________________________________
_______

5. In summary, what factors affect enzyme activity?

__________________________________________________________________________
_______
__________________________________________________________________________
_______
__________________________________________________________________________
_______
__________________________________________________________________________
_______

Conclusion:
__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
_____________________
__________________________________________________________________________
_______
 Carbohydrates
Experiment no.6
Theory

Carbohydrates include polyhydroxy aldehydes or ketones and their by-products. They

make up most of our diet and serve as the major source of energy. They play an important part
in metabolic processes by furnishing the carbon chain for compound synthesis by living
organisms. Carbohydrates can be categorized as monosaccharides, disaccharides and
polysaccharides. Monosaccharides or simple sugars are highly soluble in water, less soluble
in ethanol, and insoluble in ether. They cannot be further hydrolyzed to simpler units. They
are either aldoses and ketoses depending on the type of functional group present. They may
also be classified into tetroses, pentoses or hexoses depending on the number of carbon atoms
they possess. Free monosaccharides serve are all reducing sugars. They also exhibit
mutarotation, which means they can exist in α- and β- forms. Disaccharides are formed by two
molecules of monosaccharides. Examples of disaccharides are maltose, which are abundant
in germinating barley; sucrose, also known as cane sugar; and lactose or milk sugar, which
does not taste very sweet and is not fermented by yeast. Most polysaccharides found in nature
function either as structural units, (e.g., cellulose) or for storage such as starch, dextrin,
glycogen, and inulin.

The chemical tests used to detect the presence of carbohydrates are based on the ability
to: (a) form furfural and its derivatives; and (b) reduce and form characteristics compounds with
reagents like Tollen’s reagent and Benedict’s solution.

A. Tests based on the formation of furfural and its derivatives

Sugars are stable to hot dilute mineral acids. But hot sulfuric acid will dehydrate them
into furfural and its derivatives. Pentoses, when heated with concentrated sulfuric acid, will
form furfuraldehyde, while hexoses will form 5-hydroxymethyl furfural.

In the presence of acid, phenolic compounds like α-naphtol, orcinol, and resorcinol will
react with furfural or hydroxymethyl furfural to form colored condensation products. The
formation of these colored compounds constitutes a positive test for carbohydrates.

1. Molisch Test
The Molisch test is the general test for carbohydrates. The sugars are mixed with
α-naphtol. The test tube is inclined, and concentrated H2SO4 is added along the side of
the tube, causing the formation of a lower layer of acid. The concentrated sulfuric acid
will dehydrate the sugar allowing it to react with the alcohol forming furfural or
hydroxymethylfurfural. Formation of a purple ring at the interface of the two liquids will
indicate the presence of carbohydrate.
 2. Bial’s Orcinol test
Bial’s test is used to determine the presence of pentoses and nucleotides that
contain pentose sugars. When pentoses are treated with orcinol, furfurals are formed
and they will yield a blue-green compound in the presence of ferric ions. The reaction
is not specific for pentoses because other compounds like trioses, uronic acids, and
certain heptoses will also give blue or green products. Hydroxymethyl furfural is formed
from hexoses to give yellow-brown condensation products.

3. Seliwanoff’s Test
This test is used to differentiate ketohexoses from aldohexoses. Ketohexoses
react faster with the solution containing hydrochloric acid and resorcinol than
aldohexoses. The dehydrated ketohexoses form a bright cherry red condensation
product, while the aldohexose yields only a pale pink coloration, a negative result. In
this test, prolonged heating of samples should be avoided.

B. Tests based on the reducing property of sugars

Reducing sugars must have a free aldehyde or ketone group. Therefore all
monosaccharides and some disaccharides can reduce oxidizing reagents such as the cupric
ion, dinitrosalicylic acid, and picric acid.
1. Benedict’s Test
Benedict’s test is a very sensitive test done under mildly alkaline conditions. The
reagent contains CuSO4, Na2CO3 and sodium citrate. The formation of a brick red
precipitate of Cu22O is considered positive. Most aldehydues have the ability to reduce
Benedict’s reagent. Other compounds like formic acid, hydrazobenzene, phenols,
phenylhydrazine, pyrogallol, and uric acid will also give a positive result in this test.

2. Barfoed’s test
Barfoed’s reagent contains cupric acetate in dilute acetic acid and is used to
distinguish between monosaccharides, disaccharides, and oligosaccharides. Barfoed’s
reagent oxidizes monosaccarides, but not oligosaccharides. Disaccharides are less
oxidized but are oxidized if they undergo prolonged heating, causing hydrolysis of the
disaccharides into monosaccharides, which will then give a positive result. The
concentration of the sugar solutions used in this test should be approximately the same,
because the use of a more concentrated disaccharide solution may give a faster reaction
than that of a relatively more dilute monosaccharide solution. Unlike Benedict’s test,
Barfoed’s test is carried out under acidic rather than basic medium.

3. Phenylhydrazine test (Osazone formation)

When sugar is added to phenylhydrazine and NaAc and then heated, a yellow
precipitate is formed. The precipitate formed may then be compared with standards
using difference in melting points to identify the composition of the precipitate. Another
portion of the precipitate may be examined under the microscope to reveal the formation
of a distinctive crystalline structure. Gluccosazones are fine yellow needles aggregated
like “bundles of hay”. Note that glucose, maltose and mannose form the same
osazones. Lactosazone crystals are irregular clusters of fine needles, and look like a
“powder puff”. Maltosazone crystals are “star-shaped.

4. Tollen’s test
 Sugars with aldehyde groups are capable of reducing Tollen’s reagent (an
ammoniacal solution of Ag+) to form a gray to black precipitate. If the reaction
vessel is clean and rate of deposition is slow enough, the Ag+ deposits as a silver
mirror.

Objective
To be able to identify the different types of carbohydrates using the different specific
chemical tests.

Materials
20 test tubes dropper
test tube rack glass slides with cover
test tube holder microscope
test tube brush distilled water
aspirator and (1ml) pipette 3%solutions of glucose, xylose, fructose,
lactose,
graduated cylinder sucrose, starch
spatula Molisch reagent
cotton balls Bial’s Orcinol reagent
watch glass Seliwanoff’s reagent
water bath Benedict’s reagent
tripod Barfoed’s reagent
Bunsen burner Phenylhydrazine (solid)
Wire gauze Iodine solution
Spot plate Concentrated H2SO4
250 ml beaker Concentrated HCl

Procedures

I. Test based on the formation of furfural and its derivatives.

1. Molisch Test
a. Mix 4 ml of distilled water and 2 drops of the Molisch reagent in a test tube. This
tube will serve as the control.
b. Place 4 ml of 3% solution of glucose in a second test tube. Add 2 drops of the Molisch
reagent and mix the contents by gently shaking the test tube.
c. Incline the test tube and cautiously add about 5 ml of concentrated sulfuric acid,
allowing the acid to run down the side of the tube. Sulfuric acid is denser than water
and will form a lower layer. Note the color of the ring formed at the junction of the
two liquids.
d. In the same manner of adding acid, add sulfuric acid to the control tube. What do you
observe?
e. Repeat the above test with 3% sample solutions of glucose, lactose and starch.
f. Record all results.

Substance Tested Description of visible results

Control
Glucose
 Lactose
Starch

2. Bial’s Orcinol Test

a. Place 1 ml of 3% solution of glucose and starch in separately-labeled test tubes.
b. Add 3 ml of Bial’s reagent to each test tube.
c. Carefully heat each tube over a Bunsen flame until the solution begins to boil.
Add 1-2 drops of 10% FeCl3 solution.
d. Note the color of the product formed.
e. Record your results in the table below.

Substance Tested Description of visible results

Control
Glucose
Starch

3. Seliwanoff’s test
a. Place 1 ml each of 3% glucose, fructose and sucrose in separately-labeled test
tubes.
b. Add 4 ml of Seliwanoff’s reagent to each test tube.
c. Place the tubes in a water bath filled with boiling water and allow them to stay
there for exactly 1 minute.
d. Note the changes and record which test tube gives a positive result in the shortest
time.
e. Continue heating and observe the color changes at 1 minute intervals.
f. Record the time required for a positive test for each sample.

Substance tested Time Result Explanation

Glucose
Fructose
Sucrose

II. Tests based on the reducing property of sugars

1. Benedict’s test
a. Place 1 ml each of 3% solutions of glucose, fructose, lactose and sucrose in
separately-labeled test tubes. Add 5 ml of benedict’s reagent in each test tube.
b. Place all the tubes in boiling water bath for 2 to 3 minutes.
c. Observe the color of the solution and note whether a precipitate was formed. A
change in color of the solution is not considered a positive reaction.
d. Avoid prolonged heating.
e. Record your results.

Substance tested Result Explanation

Glucose
Fructose
Lactose
Sucrose

2. Barfoed’s test
a. Place 1 ml each of 3% solutions of glucose, fructose, lactose and starch in
separately-labled test tubes.
b. Add 3 ml of Barfoed’s reagent in each test tube.
c. Prepare a control tube using distilled water instead of sugar solution.
d. Place all the tubes in a boiling water bath for 10 minutes. Record your
observations.

Substance tested Result Explanation

Control (distilled water)
Glucose
Fructose
Lactose
Starch

3. Tollen’s test
a. Place 5 drops of 3% solutions of glucose and sucrose in separate test tubes.
b. Add 2 ml of Tollen’s reagent into each tube. (Note: Prepare Tollen’s reagent by
adding 1 drop of NaOH solution to 6 ml of 5% AgNO3. Add dilute NH4OH ( 1 ml
concentrated NH4OH + 5 ml water) until brown precipitate of silver oxide that
forms just dissolves.
c. Boil for about 5 minutes. Note and record your observations.

Substance tested Result Explanation

Glucose
Sucrose

CAGAYAN DE ORO COLLEGE - PHINMA

Biochemistry Laboratory Final Report

Name: ___________________ Rating: _____________

Course/Year/ Section: ______ Date Performed: ____________
Group no. _______________ Date Submitted: ____________

Experiment no. _____

Title: ___________________________________________________________
________________________________________________________________

Objectives: _______________________________________________________
________________________________________________________________
________________________________________________________________
________________________________________________________________
Materials: ______________ ________________
______________ ________________
______________ ________________
______________ ________________

I. Test based on the formation of furfural and its derivatives.

1. Molisch Test

Substance Tested Description of visible results

Control
Glucose
Lactose
Starch
Based on the results, which carbohydrates gave a positive result with Molisch test?
__________________________________________________________________________
__________________________________________________________________________

2. Bial’s Orcinol Test

Substance Tested Description of visible results

Control
Glucose
Starch

3. Seliwanoff’s test

Substance tested Time Result Explanation

Glucose
Fructose
Sucrose
II. Tests based on the reducing property of sugars
4. Benedict’s test

Substance tested Result Explanation

Glucose
Fructose
Lactose
Sucrose

5. Barfoed’s test

Substance tested Result Explanation

Control (distilled water)
Glucose
Fructose
 Lactose
Starch

6. Tollen’s test

Substance tested Result Explanation

Glucose
Sucrose

Questions:

1. Will disaccharides and polysaccharides give a positive result for Molisch test?
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
__________________________________________

2. Which of the different carbohydrate tests would give a positive result for maltose?
Describe the color change?
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
__________________________________________

3. Can Seliwanoff”s test be used to distinguish sucrose from fructose? Explain your answer.
__________________________________________________________________________
_______
__________________________________________________________________________
_______
__________________________________________________________________________
_______
__________________________________________________________________________
_______
__________________________________________________________________________
_______
__________________________________________________________________________
_______
4. Inulin is a polysaccharide composed entirely of fructose units. Which test should be used to
best identify the presence of fructose?
__________________________________________________________________________
_______
__________________________________________________________________________
_______
__________________________________________________________________________
_______
__________________________________________________________________________
_______
__________________________________________________________________________
_______
__________________________________________________________________________
_______

Conclusion:
__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
_____________________
__________________________________________________________________________
_______
 Hydrolysis of Carbohydrates
Experiment no. 7

Theory
Polysaccharides are complex carbohydrates of high molecular weight. They are
composed of countless glucose units combined through the loss of molecules of water. They
are different from monosaccharides and disaccharides because they do not reduce Benedict’s
solution and do not taste sweet.
The amylase fractions of starch are hydrolyzed into maltose and glucose units from the
non-reducing end by cleaving the α-(1,4) linkages. The more highly-branched fraction,
amylopectins, are degraded into a mixture of glucose, maltose and a highly-branched core
called a “limit dextrin”. Amylose gives a blue color with iodine, while amylopectin gives a purple
to red color. The color given by amylose depends on the length of the chain. The longer the
chain, the bluer the color. Purple is produced by a mixture of blue and ed complexes.
The products of hydrolysis and the color it yield with iodine are given in the table below:

Carbohydrate starch amylodextrin erythrodextrin achroodextrin Glycogen/

amylopectin
Color blue purple Red Colorless faint red*

*Cooling the solution in ice water will intensify the color.

Objectives
1. To differentiate the color given by polysaccharides in the iodine test.
2. To identify he hydrolysis products of polysaccharides.

Materials
Water bath iron stand
Beaker pipette
Stirring rod Benedict’s solution
Spot plate iodine solution
Graduated cylinder HCl
Iron ring 3% starch solution

Procedures
I. Hydrolysis of polysaccharides
 Place 30 ml of 3% starch solution in a small beaker. Acidify by adding 20 drops of
concentrated HCl. Cover the beaker containing the mixture with a watch glass and boil gently
in a water bath. As soon as the mixture becomes translucent, get 2 drops of the starch solution
and place in a spot plate. Test the spot by adding one drop of iodine solution. Place the other
drop in a test tube and test sing Benedict’s solution. Repeat withdrawing samples of the
mixture every two minutes. Continue heating the solution until you have repeated this
procedure ten times. Record your results on the table below by rating the color using 10 as
the darkest and 1 as the lightest. Indicate any changes in color.

Test reagent 1 2 3 4 5 6 7 8 9 10
Iodine solution
Benedict’s solution

CAGAYAN DE ORO COLLEGE - PHINMA

Biochemistry Laboratory Final Report

Name: ___________________ Rating: _____________

Course/Year/ Section: ______ Date Performed: ____________
Group no. _______________ Date Submitted: ____________

Experiment no. _____

Title: ___________________________________________________________
________________________________________________________________

Objective: _______________________________________________________
________________________________________________________________
________________________________________________________________

Materials: ______________ ________________

______________ ________________
______________ ________________
______________ ________________

Test reagent 1 2 3 4 5 6 7 8 9 10
Iodine solution
Benedict’s solution

Questions:
1. Explain why the iodine test gave such results upon prolonged heating.
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
 _____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
____________________________________

2. Why should a very dilute solution of iodine be used for this test?
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
__________________________________________

Conclusion:

______

Effect of Antibiotics on Lactic Acid Fermentation

Experiment no.8

Theory
The metabolic cycle that oxidizes glucose (the absorbable form of carbohydrates) to
pyruvate under anaerobic condition is called glycolysis, or the Embden Meyerhof Scheme
(EMS). If anaerobic condition persists, pyruvate is reduced to lactate. This reaction is
catalyzed by lactic acid dehydrogenase (LDH), with NADH as coenzyme. This process
releases energy, which is utilized by some microorganisms to change milk into yoghurt. This
change is visibly manifested by the “curdling” of milk. The process is affected by drugs,
particularly antibiotics.

Objective
To determine the effect of different antibiotics on lactate fermentation.

Materials
Antibiotic capsules (500 mg) sterile beaker (cover for milk can)
30 sterile test tubes Yoghurt
2 (5 ml) pipet and aspirator 3 test tube racks
distilled water evaporated milk
disposable syringes 25 cotton plugs (thread, bandage gauge,
cotton)
can opener Bunsen burner
tripod wire gauze
2 graduated cylinder, 10 ml disinfectant
rubbing alcohol scissor
masking tape
 Note: All glasswares used for this experiment must be sterile.
Procedures
1. Preparation of antibiotic solutions
Using a pipet, prepare antibiotic solutions of four different concentrations.

2. Inoculation of 25 samples of milk with bacteria

a. Wipe the top of the can of milk with disinfectant for at least two minutes.
b. Puncture the can with a can opener that has been sterilized over a blue flame. When
not in actual use, cover the can with a sterile beaker.
c. Prepare 25 test tubes and provide with cotton plugs. Sterilize a clean pipet over a
blue flame and use it to fill each of the 25 sterile test tubes with 3 ml of milk.
Remove the cotton plugs just before filling and replace immediately after filling.

Note: The inoculation procedure with bacteria (from yoghurt) should be dome aseptically as
follows:
a. Bring a filled test tube close to the flame of an alcohol burner. Remove the cotton
plug and pass the entire rim of the test tube over the blue flame thrice.
b. Heat the tip of a dropper over the blue flame and use it to draw the yoghurt. Add four
drops of yoghurt to each of the milk samples. Do not allow the drops of yoghurt to
roll down the sides of the tube.
c. Pass the rim of the test tube over the blue flame again and replug with cotton before
treating the next test tube.

3. Addition of antibiotic solution

a. Label all test tubes with the proper replicate numbers (1 to 25) and assigned
experimental treatment.
b. Set aside 5 control tubes. To the rest, add 8 drops of the assigned antibiotic solution.
c. Swirl the tubes gently after each addition.

4. Incubation of the samples

a. Arrange the experimental tubes in randomized complete block design in test tube
racks. All experimental units of a single replicate (referred to as a block) are placed
together but randomly arranged within the blocked area.
b. Store the entire setup in a clean laboratory space. Label with your schedule, section
and group number.
c. Check for milk curdling daily over a period of three days.
d. Discard all samples by wrapping in an envelope and throwing them in the trash can
for wet garbage.

Data and Results

Record observation using the following code:
Write “0” if there is no curdling
Write “1” if an appreciable degree of milk curdling was observed.

Antibiotic Used _____________

 Replicate Number Day 1 Day 2 Day 3
1
Control 2
3
4
5
1
C1 2
3
4
5
1
C2 2
3
4
5
1
C3 2
3
4
5
1
C4 2
3
4
5

CAGAYAN DE ORO COLLEGE - PHINMA

Biochemistry Laboratory Final Report

Name: ___________________ Rating: _____________

Course/Year/ Section: ______ Date Performed: ____________
Group no. _______________ Date Submitted: ____________

Experiment no. _____

Title: ___________________________________________________________
________________________________________________________________

Objective: _______________________________________________________
________________________________________________________________
________________________________________________________________
Materials: ______________ ________________
______________ ________________
______________ ________________
______________ ________________

Data:
Replicate Number Day 1 Day 2 Day 3
1
Control 2
3
4
5
1
C1 2
3
4
5
1
C2 2
3
4
5
1
C3 2
3
4
5
1
C4 2
3
4
5
Questions:

1. Based on the results, what is the effective concentration of antibiotic that will prevent
curdling?
_____________________________________________________________________
_____________________________________________________________________
____________

2. What is lactic acid fermentation? Show the reactions involved in the process.
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
__________________________________________

3. What type of microorganisms carry out the fermentation of milk to yoghurt?

_______________________________________________________________________
_______
_______________________________________________________________________
_______
_______________________________________________________________________
_______

4. The word antibiotic comes from antibiosis. What is antibiosis?

_______________________________________________________________________
_______
_______________________________________________________________________
_______
_______________________________________________________________________
_______
_______________________________________________________________________
_______

5. How did the antibiotics used interfere with the process of fermentation in this experiment?
_______________________________________________________________________
_______
_______________________________________________________________________
_______
_______________________________________________________________________
_______
_______________________________________________________________________
_______
_______________________________________________________________________
_______
_______________________________________________________________________
_______

6. Why is it absolutely necessary that aseptic conditions be maintained throughout the

experiment?
_______________________________________________________________________
_______
_______________________________________________________________________
_______
_______________________________________________________________________
_______

7. If a sterile environment was not maintained in the procedures of this experiment, what
would have happened? Did this happen in your setup?
_______________________________________________________________________
_______
 _______________________________________________________________________
_______
_______________________________________________________________________
_______
_______________________________________________________________________
_______

Conclusion:
__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
___________________________________