Philippine College Of Criminology

Natural Science Department

General Laboratory Rules and Safety Precautions

The Chemistry Laboratory can be a place of discovery and learning. However,

considering the nature of work done in the laboratory, it can be a dangerous place if safety
precautions are not taken. Students are expected to learn and follow the general working
guidelines in the laboratory to ensure a safe environment for them and the people they may
be working with additional safety precautions will be announced in class prior to experiments
where a potential danger may exist.

Attire

1. Safety goggles and laboratory gowns/aprons must be worn whenever you work in the
lab. Gloves should be worn whenever you use chemicals that may cause skin
irritations or when you need to handle hot equipment. Contact lenses are not allowed
unless you have permission from your instructor. Even when worn under safety
goggles, various fumes may accumulate under the lenses and cause serious injuries or
blindness.
2. Dress properly during a laboratory activity. Long hair, dangling jewelry and loose or
baggy clothing are hazardous in the laboratory.
3. Closed toe shoes and long pants must be worn in the lab. Sandals and shorts are not
allowed.
4. Long hair must be tied back when working with flames.

Personal Protective Equipment (PPE)

1. The use of a laboratory coat (buttoned closed and properly fitted) is required when
working in the laboratory. Students will not be allowed to conduct practical without a
laboratory coat.
2. Students are required to provide their Personal Protective Equipment (PPE) such as
fluid resistant gowns, gloves, goggles, and face masks. These PPE will be used when
there is significant probability that potentially hazardous substances may be splashed
on the student.
3. Protective clothing must be removed before leaving for non-laboratory areas.

Conduct Inside the Laboratory

1. Be familiar with your lab assignment before you come to the lab premises. Follow all
written and verbal instructions carefully. If you do not understand a direction or a part
of a procedure, ask the instructor before proceeding with the experiment.

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2. Do not touch any equipment, chemicals, or other materials in the laboratory until you
are instructed to do so.
3. Eating, drinking and smoking are strictly prohibited in the laboratory. Do not use
laboratory glassware as container for food or beverages.
4. No unauthorized experiments are to be performed.
5. Never taste anything.
6. Never directly smell the source of any vapor or gas. Instead, through your cupped
hand, waft a small sample to your nose.
7. Coats and backpacks, among others should not be left on the lab tables and stools.
Lab chemicals can destroy personal belongings.
8. Observe good housekeeping practices. Work areas should be kept clean at all times.
9. Know the locations and operating procedures of all safety equipment including the
first aid kit, eyewash station, safety shower, spill kit, fire extinguisher and fire blanket.
Know where the fire alarm and the exits are located.
10. Be alert and always proceed with caution in the laboratory. Notify the instructor
immediately of any unsafe condition you observe.
11. Dispose all chemical wastes properly. Never mix chemicals in sink drains. Sinks are
to be used only for water and those solutions designated by the instructor. Solid
chemicals, metals, matches, filter paper and all other insoluble materials are to be
disposed of in the proper waste containers, not in the sink. Check the label of all
waste containers twice before adding your chemical waste to the container. Cracked
or broken glass should be placed in the special container for it.
12. Labels and equipment instructions must be read carefully before use. Set up and use
the prescribed apparatus as directed in the laboratory instructions provided by your
instructor.
13. Keep your hands away from your face, eyes, mouth and body while using chemicals.
Wash your hands with soap and water after performing all experiments. Clean (with
detergent powder), rinse, and dry all work surfaces after the end of the experiment.
14. At the end of the laboratory session, ensure that a) the main gas outlet valve is shut
off; b) the faucet is turned off; c) desk top, floor area, and sink are clean; and d) all
equipment are cool, clean and arranged properly.

Proper Handling of Chemicals and Equipment

1. Consider all chemicals to be hazardous unless you are instructed otherwise.

Material Safety Data Sheets (MSDS) are available in the lab for all chemicals in
use. This will inform you to any hazards and precaution.

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2. Know what chemicals you are using. Carefully read the label twice before taking
anything from the bottle. Excess reagents are never to be returned to stock bottles.
If you have taken too much, dispose off the excess.
3. Many common reagents, like, alcohol and acetone, are highly flammable. Do not
use them anywhere near open flames.
4. Always pour acids into water. Water poured into acid creates a heat reaction that
would cause the water to explode into steam, sometimes violently, which in turn
would cause the acid to splatter.
5. If chemicals come into contact with your skin or eyes, flush immediately with
abundant amounts of water and consult with your instructor.
6. Contact the stockroom technician for clean-up of mercury spills.

In Case of Accidents:

The following are First Aid treatments:

A. FIRE

1. Small fires such as in the beaker or in the flask:

First, turn off the gas outlet. Then attempt to smother the fire with an asbestos pad or wet
towel. If this fails, use the fire extinguisher provided for this purpose.
2. Clothing on fire
Smother fire by covering yourself with wet blanket or by rolling on the floor.
3. Phosphorous or Sodium Fire
Put out fire by throwing sand on it. In general, do not use water, this will only result in
the spreading of the fire.

B. CUTS

First, wash with water thoroughly and then apply 70% alcohol or tincture of iodine.
Bandage with sterile gauze. Do not continue to use iodine in subsequent dressing, burn will
result. Never cover directly with adhesive tape.

C. CHEMICAL SPILLS

1. Acid in the eye

Wash thoroughly with running water, then by means of a cap bathe with 2% sodium
bicarbonate solution. Dry with sterile gauze and put several drops of olive oil.

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2. Alkali in the eye

Wash thoroughly with running water, and then bathe with saturated solution of boric acid.
Dry with sterile gauze and drop olive oil into the eye.
Never use a strong acid or a base to neutralize each other on your clothing or body.

D. BURNS

Immediately plunge the burned areas with cold water for several minutes.

1. Acid burns

Wash with running water then with saturated sodium bicarbonate solution. Cover for
about ten minutes with solid sodium bicarbonate. Wash off, dry with sterile gauze and
visit the hospital for further treatment.
2. Alkali burns

Wash with running water then with saturated boric acid solution. Cover for about ten
minutes with powdered boric acid. Wash off, dry with sterile gauze and apply carbonated
vaseline.
3. Heat burns

Apply Vaseline, butesine picrate, or ask for a burn ointment from your lecturer or
laboratory assistant.

E. LIQUID SPLASH ON THE EYE

Wash the eye immediately with water from the eyewash bottle or eyewash fountain.

F. CHEMICAL SPLASH ON THE SKIN

Immediately rinse the area with cold water for at least one minute. Notify your lecturer
for further action.

REPORTING OF ACCIDENTS/INCIDENTS
1. All injuries or unusual incidents must be reported immediately to the lecturer or
laboratory staff.
2. All accidents/incidents must be documented by the lecturer or laboratory staff using
the incident report form.

BEFORE LEAVING THE LABORATORY:

1. Clean your work area at the end of the laboratory session.

2. Be sure that your water and gas outlets are turned off.
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3. Remove your laboratory coat.
4. Wash and dry your hands.

COMMON LABORATORY TECHNIQUES

A. Measuring volume

The graduated cylinder, volumetric flask, burette and pipette are used to measure liquid
volume in the laboratory. When measuring volume with these devices, use the lower
meniscus for transparent liquids and the higher meniscus for opaque/colored liquids. Read
the same point in the meniscus consistently for a given liquid. Make sure that the apparatus is
held vertically straight and the eye is at the same level as that of the meniscus. Your slips of
vision should be exactly perpendicular to the scale to avoid false reading caused by parallax.

Figure 1. Reading a Meniscus

1. Graduated cylinder: A graduated cylinder is generally used for approximate
measurements with accuracy and not greater than 0.5%. Graduated cylinders of
different capacities are available. It is recommended that the volume of the
cylinder should not be more than ten times the volume to be measured for better
accuracy.

2. Burette: Burettes are used to accurately measure volume in titration. There are
two kinds of burette: a) an acid burette with a glass stopcock; and b) a base
burette with a rubber connection bearing a glass delivery tip and pinchcock.

3. Pipette: Pipettes are used for accurate measurements of volumes to be transferred

from one vessel to another.

Techniques in Handling a Pipette

a. When using a pipette, always use a rubber aspirator. Before attaching the aspirator
to the pipette, squeeze to remove the air inside and then release.

b. Rinse the pipette with the liquid to be measured before use. Put a small
amount of the liquid to be measured into a clean and dry beaker. Without
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attaching the bulb into the stem, hold it firmly against the end of
the pipette stem as you squeeze (this will make the removal of the bulb easier).
Dip the tip of the pipette into the liquid and gradually release the bulb to draw
only 1 or 2mL of the liquid into the pipette. Rotate or move the pipette in a near
horizontal so that the near inner surface comes in contact with the liquid. Drain
the liquid.

c. After rinsing, dip the tip of the pipette in the liquid. Draw the liquid after
squeezing the air out of the aspirator. Gradually release the aspirator so that the
liquid rises slowly in the pipette.

d. Remove the excess liquid by holding the pipette vertically, letting the air enter
by using the index finger to allow the liquid to flow out slowly into the
beaker .Do this until the meniscus coincides with the calibration mark.

e. Insert the tip of the pipette well inside the receiver. Hold the pipette vertically and
tilt the receiver so that the pipette’s tip touches the wall of the receiver. Allow the
liquid to flow freely down the wall of the receiver. When free flow stops, keep the
pipette in contact with the wall for 15seconds more so that the pipette walls will
have a chance to drain. Do not shake the pipette or blow out the liquid left at its
tip.

Figure 2. Proper Handling of the Pipette

B. Determining Mass

Three commonly used weighing scales are found in the laboratory. These are:

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1. Triple Beam Single Pan Balance: This balance is used for
semi-micro masses and has a maximum capacity of 111grams. The middle
scale reads up to 100 grams in 10 grams notched steps, the rear scale up to
10grams in 1 gram notched steps, and the front scale up to 1 gram with the
rider sliding over a scale graduated into 0.01 units.
2. Platform Balance: The single beam is graduated for 10grams in 0.1gram
divisions. The balance is sensitive to 0.1gram and can read up to 10gram
without additional masses.
3. Electronic Top-Loading Balance: This can weigh up to three decimal places
or up to the nearest thousandth of a gram.

Determining the Mass of an Object Using a Platform Balance

a. Clean the pans with dry cloth.
b. Place all the masses on the beam at zero position.
c. Make sure that the pointer swings equally to the left and to the right of the
scale or it stays at zero position.
d. Place the object to be weighed on the left pan. Never place a chemical
directly on the pan.
e. Always start with heavier rider. If the right pan is still higher than the left
pan, it means that the object is heavier than the mass on the beam. Move
the rider to the next notch until the right pan becomes lower than the left
pan.

Safety Precaution when Using the Balance

a. When transferring the balance from the cabinet to your table, carry it with
your left hand supporting the base and your right on the pointer support.
b. Place powdered or granulated solids to be weighed on a piece of paper, the
mass of which have been previously determined.
c. Wipe off immediately any spilled chemicals with a clean rug. Keep the
balance away from the supply of reagents and from corrosive fumes.
d. Avoid jarring the balance by roughly adding or subtracting a weight or
weights.
e. After using the balance, replace all the sliding weight on the zero mark.

C. Measuring Temperature

A thermometer is a device for measuring temperature. It has a bulb that contains mercury

attached to a fine graduated glass capillary.

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Graduated in Celsius degree, the zero (0o) mark corresponds to the
freezing temperature of water at a pressure of one atmosphere (atm). The 100o mark is the
normal boiling temperature of water. The column length between 0o and 100ois divided into
100 equal distances; each scale division is equal to one degree Celsius.

How to Make Temperature Readings

When reading the thermometer, your eyes must be at same level as the mercury
column.

Safety Precaution when using the thermometer

a. Do not hold the mercury bulb of the thermometer. This end is prone to breaking.
b. In case the thermometer breaks, collect the mercury glass by passing a cooper sheet
or wire over each droplet. Place the collected mercury in the bottle. Add enough water
to submerge the mercury. Cover tightly.
c. All accidents involving mercury must be reported to the teacher.
d. Do not play with mercury. Mercury is volatile and its vapor is poisonous.
e. To insert the thermometer into a cork/rubber, stopper, lubricate the thermometer and
stopper with water or little oil. Hold the thermometer with a piece of cloth near the
end. Insert using a twisting motion. This would prevent the breaking of the
thermometer at the end of the stress.

Dispensing of Chemicals
A. Handling Bottles Containing Liquid

1. When pouring chemicals from glass-stoppered reagent bottles, remove the stopper
and hold it in your fingers while carefully pouring the liquid into the desired
container.
2. When pouring form a screw cap bottle, set up the cap upside down on the table top so
that it does not become contaminated. Be sure to put the correct cap on the bottle after
you have used it.
3. Always pour the liquid from a bottle on side opposite the label, especially if the bottle
has paper label. If you spill any liquid or drip some on the side of the bottle, clean it
up.
4. Never put the chemicals back on the reagent bottles to prevent contamination.

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a) Glass-stoppered reagent bottles b) Screw cap bottle

c) Transferring liquid from a reagent bottle

Figure 3a, b and c. Proper Handling of Reagent Bottles

B. Dispensing Solids from Bottles

When dispensing samples of powdered crystalline from a jar, pour the desired amount of
the solid on a small piece of clean paper or into a clean beaker. Carefully tilt the jar and rotate
it back and forth to work the solid up to the mouth. Then using the same back and forth
rotation, allow the desired amount of solid to fall from jar. Be careful when transferring a
solid from a jar. Never put any solid back into the jar. Never put wooden slits, spatulas, or
paper into a jar of solid unless your instructor allows it. Solid may be poured into test tubes
by using a piece of paper that has been creased down at the center.

C. Heating in a Test Tube

Hold the test tubes with a test tube holder in a slanting position or at 45 o position. Move
the tube gently along the length so that heat would be evenly distributed. Never point a test
tube (or any beaker that you are heating) at yourself or anyone else. Do not heat the test tube
directly at the bottom as this may cause the liquid to suddenly boil and erupt out of the tube.

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Figure 4. Heating in a Test Tube

D. Using an Evaporating Dish

An evaporating dish is a shallow, heat-resistant porcelain ware. It is used when

evaporating volatile solvent from a solution, leaving behind a solid.

Figure 5. Evaporation of a Solution

How to Evaporate Solutions

Heat is applied directly to the evaporating dish with or without wire gauze (direct
heating may be with an alcohol burner). The solvent is allowed to evaporate leaving the

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solute on the dish. A gas flame is not recommended for evaporating small
amount of solution

because spattering may occur with high temperature. Also, some solids decompose at high
temperature.

Safety Precautions when Using the Evaporating Dish

 It is always safe to evaporate all liquids through steam bath.
 When evaporation is done using direct heat, reduce the flame when only a small
amount of liquid is left so as not to decompose the residue.
 Let the dish cool down before removing the solid.
 Use a cloth pad to remove the dish from the setup.
 Do not put the hot porcelain on a wet or cold surface to prevent breakage due to
sudden change in temperature.

E. Preparing Filter Paper

Fold the paper along its diameter and fold it again to a quadrant. Tear off the smaller
quarter and open it into the shape of a cone with one thickness on one side and three layers
(two with the torn corner) on the other side. The torn corner prevents an air column from
existing between the glass funnel and the filter. Adjust the cone to fit the funnel exactly.
Moisten the filter paper with distilled water and press evenly until there are no air bubbles
between the filter paper and the funnel.

Figure 6. Proper folding of Filter Paper

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Exercise No. 1
The Big Picture

Objective:
1. To demonstrate cooperation among members to finish an assigned task within a
given period of time.
2. To build leadership among members of a certain group.
3. To focus and understand solving a situation assigned by the instructor.

Introduction:

This activity can serve many purposes. It can serve as an icebreaker at the
beginning of a course. It can be used to reveal the prominence of organic chemistry
in everyday life. It can be used as a jumping point for group work or other
collaborative activities in a class. Finally, this has been done as a fun activity to
motivate the students in their learning the concepts of Organic Chemistry.

Instructions:

1. A series of illustrations will be given to each group.

2. Each successive picture focuses on one detail from the previous picture.
3. Along with scenes from everyday life, each illustration contains some snippet of
organic chemistry such as a molecule from an object in the scene, a reaction from
the course, or a key concept.
4. Aside from organic chemistry, the series of illustrations have another thing in
common and this will be a competition to see which group can discover this common
thread first. Remember that a group cannot visually reveal their assigned
illustration to another group. Rather, they are only allowed to describe the
illustration verbally to one another in their own group.
5. Care is taken to randomize the illustrations prior to distribution.
6. This activity takes approximately 5 minutes after each group has been assigned a
picture. In general the group dynamics develop slowly, with each member turning to
his or her co-member and inquiring about the contents of the other’s illustration.
In some groups this process, in which one student queries another and then moves
on to the next student, continues for several minutes. Eventually, each member
describes his or her illustration to the entire group. Eventually one member will be
the responsible to record what has been explained by his member regarding the
picture until all twelve (12) pictures are accounted for. As member in the group
begin to realize that each illustration contains an object from another illustration, the
subgroups become larger until the common thread is solved. The ultimate “winner”
is the group that correctly arranges itself in the order of the pictures as your

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instructor will be the one to declare it.

Discussion:

It is interesting to note the variations among the groups. In some, leaders

become apparent as they try to corral other members of the group to become more
organized. In other groups, a sole leader will emerge that directs the actions of the
whole. Still other groups never reveal true leaders, as members go about the activity in
a seemingly random fashion. Your instructors may take note of any leaders that
emerge, as this can aid in the assigning of small groups for future collaborative
classroom activities.

This technique is usually used at the beginning of a semester. Because of the

chemical content of each picture, one might expect the nature of the activity to vary
depending on the level of organic chemistry knowledge of the group. To solve the
puzzle, it is almost better to not know any organic chemistry (i.e., to do the activity
at the beginning of the semester), so that students do not dwell too much on the
organic structures and concepts of each picture. In any event, the activity creates a
situation in which every group member is required to contribute to the conversation.

From Thomas Poon concepts here are the following pictures:

A. Carbon atom B. Crude Oil Factory

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C. Helicopter D. Family watching television

E. Propane molecule F. National highway

G. Hot air balloon H. Carbon chain molecule

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I. Residential community J. Man in an airplane

K. Cruise ship L. Airplane landing

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DATA SHEET
Activity No. 1
The Big Picture

Name: _____________________________ Rating: ____________________

Section:____________________________ Instructor: _________________
Schedule: __________________________ Date: _____________________

Explain briefly:

Picture 1: ________________________________________________________

Picture 2: ________________________________________________________

Picture 3: ________________________________________________________

Picture 4: ________________________________________________________

Picture 5: ________________________________________________________

Picture 6: ________________________________________________________

Picture 7: ________________________________________________________

Picture 8: ________________________________________________________

Picture 9: ________________________________________________________

Picture 10: _______________________________________________________

Picture 11: _______________________________________________________

Picture 12: _______________________________________________________

Conclusion(s):

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Exercise No. 2
Making Molecular Models

Objective:
1. To demonstrate cooperation among members to finish an assigned task within a
given period of time.
2. To learn how to make molecular models.
3. To determine the hybridization of atoms.

Materials:

Molecular models, pencil, colored pens or crayons, sticks and balls, pair of scissors,
glue

Introduction:

Methane (CH4) is said to be the simplest organic compound or hydrocarbon.

Therefore, illustrating its molecular orbital is simple. The molecular description of bonding
in methane does several things for us.

It should reconcile our valence-bond idea of electrons localized between carbon

and hydrogen with the "delocalized" picture typical of the molecular orbital approach.
It should tell us (quantitatively) about the energies of different electrons. This energy
description should reproduce experimental findings that two of the valence electrons
have a lower energy than the other 6.

Figure 1. Molecular model of methane gas

Procedure:

Ask from your instructor what organic compound you are going to illustrate or
make a model of it. Place your model in a ¼ illustration board. In your data sheet,

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draw the structural formula of your organic compound and identify
the hybridization of each atom in the compound.

DATA SHEET
Activity No. 2
Making Molecular Models

Name: _____________________________ Rating: ____________________

Section:____________________________ Instructor: _________________
Schedule: __________________________ Date: _____________________

Draw the structural formula of your organic compound and identify the
hybridization of each atom.

Conclusion(s):

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Experiment No. 1
Melting Point of an Organic Compound

Objective:
1. To determine the melting point of a known organic compound.

Materials/Chemicals:
Powdered naphthalene, liquid paraffin or oil in a 100ml beaker, capillary tubes (10cm
long), thread, glass plate, thermometer, iron stand with clamp, stirrer, spatula, hot plate or
electric stove

Discussion:
The melting point of a solid is defined as the temperature at which the solid
exists in equilibrium with its liquid under an external pressure of one atmosphere.
On heating a solid, each molecule absorbs energy in the form of heat. As a result,
the force of attraction between the molecules decreases as the molecules become
increasingly separated. At a particular temperature called the melting point of the
solid, the separation of the molecules increases by a large amount and the solid
melts and is converted into liquid. In this experiment, students have to determine
the melting point of a given organic compound called naphthalene.

Procedure:
1. Take one piece capillary tube and seal one end in a Bunsen burner.
2. Place the powdered naphthalene in the sealed capillary tube at least 1/3 of its length.
Tap the capillary tube in order to pack the powdered naphthalene with no air gaps
between the substance.
3. Attach the capillary tube to a thermometer using a thread.
4. Take the 100 ml containing liquid paraffin or oil and place it in a hot plate of
electric stove.
5. Clamp the thermometer inside the beaker and immerse the capillary tube
containing the powdered naphthalene.
6. Start heating the liquid paraffin or oil and stir the liquid to ensure uniform heating
inside the beaker.
7. Note the temperature T 1 when the naphthalene starts melting. Continue heating
and note the temperature T 2 when the naphthalene in the capillary tube completely
melted.
Note: The average of the two temperatures T 1 and T2 is the melting point of the
naphthalene.
Precautions: Use dry and powdered naphthalene for this experiment. Slowly
heating is required to ensure gently heating of the substance. The bulb of the
thermometer and the capillary attached to it should not the sides of the bottom of
the beaker.

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DATA SHEET
Experiment No. 1
Melting Point of an Organic Compound

Name: _____________________________ Rating: ____________________

Section:____________________________ Instructor: _________________
Schedule: __________________________ Date: _____________________

Results:

T1 = ________oC
T2 = ________oC
Average T = ________ oC

Computation:

Questions:
1. What is the molecular and structural formula of naphthalene?

2. Give some uses of naphthalene?

3. What type of organic compound is naphthalene?

Conclusion(s):

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Experiment No. 2
Boiling Point of an Organic Compound

Objective:
1. To determine the boiling point of a known organic compound.

Materials/Chemicals:
Toluene, liquid paraffin or oil in a 100ml beaker, 1 pc small test tube, capillary tubes,
thermometer, dropper, rubber band, iron stand with clamp, stirrer, hot plate or electric
stove and Bunsen burner

Discussion:
The boiling point of an organic liquid is defined as the temperature at which the
vapor pressure of the liquid becomes equal to the atmospheric pressure exerted
upon the liquid surface. At one standard atmospheric pressure (1 atm), the boiling
point of the liquid is termed as the normal boiling point. Different liquids have
different boiling points.
Compound Boiling point Intermolecular force
Butane o
-1 to 1 C Van der Waal’s dispersion force
Diethyl ether o
35 C Dipole-dipole interaction
n – Butanol o
117 C Hydrogen bonding
Sodium n – butoxide >260 Co Ionic interactions

This experiment is aimed to determine the boiling point of toluene.

Procedure:

1. In your small test tube, with the use of a dropper fill it with toluene more than half of the
test tube.
2. Attach the thermometer to the test tube with the use of a rubber band in such a way
that the bottom of the test tube is at the middle of the thermometer bulb.
3. Place the beaker containing liquid paraffin or oil in a hot plate or electric stove.
4. Clamp the thermometer inside the beaker and lower the test tube into the liquid
paraffin or oil bath. Adjust the set-up so that the bulb of the thermometer is well
under the liquid bath.
5. Take one capillary tube and seal one end in a Bunsen burner. Then place the
capillary tube inside the test tube containing toluene so that its open end is dip in
toluene.
6. At first, a bubble or two will be seen escaping from the end of the capillary tube
dip in toluene but sooner rapid stream of air bubbles escape from it. At this stage,
the vapor pressure of the liquid just exceeds the atmospheric pressure.

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7. Note the temperature T 1 when continuous stream of bubbles start coming out.
Stop heating and note the temperature T 2 when the evolution of bubbles from the
end of the capillary tube just stops. The mean or average of two temperatures
gives the boiling point of the liquid.

Precautions:
 The capillary tube should be properly sealed.
 The rubber band should be fixed at the mouth of the test tube so that the
open end of the test tube with the rubber band is sufficiently outside the
liquid paraffin bath.
 The liquid paraffin bath must be heated very slowly and the bath is
stirred gently to ensure uniform heating.
 The open end of the capillary tube should be well inside the liquid.

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DATA SHEET
Experiment No. 2
Boiling Point of an Organic Compound

Name: _____________________________ Rating: ____________________

Section:____________________________ Instructor: _________________
Schedule: __________________________ Date: _____________________

Results:

T1 = ________oC
T2 = ________oC
Average T = ________ oC

Computation:

Questions:
1. What is the molecular and structural fomula of toluene?

2. Give some uses of toluene?

3. What type of organic compound is toluene?

Conclusion(s):

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Experiment No. 3
Isolation of Chlorophyll Pigment from Spinach

Objective:
2. To extract the presence of green pigment called chlorophyll from spinach.

Materials/Chemicals:
Spinach leaves, Magnesium sulfate (MgSO4), sand, sodium sulfate (Na2SO4),
petroleum ether, acetone, mortar and pestle, 3 pcs small test tube, capillary tubes,
thermometer, pipet/dropper, 50 ml beaker, hot plate or electric stove and watch glass

Discussion:

Chlorophyll is the green pigment responsible for the color of leaves. Its
presence in leaves is crucial for photosynthesis. Photosynthesis can be defined as the
process by which plants, algae, and photosynthetic bacteria use light energy to drive
the synthesis of organic compounds. The photosynthetic process involves the
removal of CO2 from the atmosphere, which is used to synthesize carbohydrates, and
results in the release of O2. The energy to drive the chemical reactions of
photosynthesis comes from the sunlight absorbed by the chlorophyll molecules.
Hence, the first step in photosynthesis is the absorption of visible light from the sun by
chlorophyll molecules. The chlorophyll molecules then transfer the light energy to
chloroplasts, the reaction center of photosynthesis. In this way light energy is
converted to chemical energy for converting CO2 into carbohydrates. The overall
reaction for photosynthesis is:
6CO2  6H2O  h  C6 H12O6  6O2
This overall equation for photosynthesis is deceptively simple. In fact, a series of
complex reactions must occur in a coordinated manner for the synthesis of
carbohydrates. To produce a sucrose molecule, C6H12O6, plants require about 30
distinct proteins that work within a complicated membrane structure.

Without light photosynthesis cannot take place; and hence the absorption of
light by chlorophyll is the first step in photosynthesis.

Chlorophyll a Chlorophyll b

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Procedure:

1. Weigh 3 grams of spinach and 3 grams of MgSO4 separately in a sheet of

paper.
2. Place the frozen spinach and MgSO4 and add a little amount of sand in a
mortar and pestle.
3. Grind the mixture thoroughly in a mortar and pestle.
4. Transfer the ground spinach into a test tube.
5. Add 5 ml of acetone to the test tube and cover it with a cork. Shake it for 5
minutes.
6. Transfer the liquid into a new test tube using a pipet.
7. To the liquid, add 4 ml of petroleum ether and 1 ml of water.
8. Again, cover the test tube and shake it for 1 minute.
9. Centrifuge it for 2 minutes.
10. Separate the colored layer and place it in a new test tube,
11. Heat it in warm water whose temperature is 60oC. The liquid boils at this
temperature, Continue heating until less than 1mL of liquid remains in your test
tube.
12. Get a strip of filter paper and draw a line ¼ inch from the bottom.
13. Using capillary tube, place a spot of the liquid from the line and then place it in
a small beaker containing 10 ml of 70:30 ether/acetone mixture. Cover the
beaker to prevent the solvent from evaporating. When the green pigment stop
to move upward, get the filter paper.
14. Dry your filter paper (chromatogram) and attach in your data sheet.

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DATA SHEET
Experiment No. 3
Isolation of Chlorophyll Pigment from Spinach

Name: _____________________________ Rating: ____________________

Section:____________________________ Instructor: _________________
Schedule: __________________________ Date: _____________________

Results:

Chromatogram

Questions:

1. How many color(s) are there in your chromatogram? What are the colors?

2. What is the purpose of petroleum ether and acetone mixture in this experiment?

3. What is the importance of chlorophyll in plants?

Conclusion(s):

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Experiment No. 4
Hydrolysis of Starch By α - Amylase

Objective:
1. To extract the presence of green pigment called chlorophyll from spinach.

Materials/Chemicals:
1% starch solution, phosphate buffer (pH = 6-8), iodine solution, 5 pcs test tube,
thermometer, 4 pcs 100 ml beaker, hot plate or electric stove, dropper and watch glass

Discussion:

Hydrolysis is any chemical reaction in which a molecule of water ruptures one or

more chemical bonds. The term is used broadly for substitution, elimination, and
fragmentation reactions in which water is the nucleophile.
Biological hydrolysis is the cleavage of biomolecules where a water molecule is
consumed to effect the separation of a larger molecule into component parts. When
a carbohydrate is broken into its component sugar molecules by hydrolysis
(e.g. sucrose being broken down into glucose and fructose), this is
termed saccharification.
Generally, hydrolysis or saccharification is a step in the degradation of a
substance.

Starch is mainly composed of two parts:

- Amylose: a linear polymer made up of α-1-4 bound D-glucose units. This
polysaccharide is making up approximately 20-30% of the structure.
- Amylopectin: a polysaccharide with highly branched polymer of α-1-6 bound
glucose units.

Amylase is an enzyme that hydrolysis alpha-bonds of large alpha-linked

polysaccharides such as starch and glycogen, yielding maltose and dextrin. It is the
major form of amylase found in humans and other mammals. Amylase is found in
saliva. This form of amylase is also called “ptyalin”. The enzyme randomly affects
α-1-4 bonds, belonging to the amylose structure of starch, and maltose units form.
Amylase does not affect the α-1-6 bonds that belong to the amylopectin structure of
starch. As a result of hydrolysis carried out with α- amylase, besides the maltose and
glucose units, a large branched dextrin structure is formed in the media.

The optimum conditions for α- amylase:

- Optimum pH: 5.6-6.9
- Human body temperature: 370C
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- Presence of certain anions and activator:

 chloride and bromide- most effective
 iodide- less effective
 sulfate and phosphate - least effective

Procedure:

1. Add 1 ml of 1% starch solution and 1 ml phosphate buffer solution (pH = 6.8)

into 4 test tubes.
2. Place each test tube into water baths at 0 0C, 250C, 370C and 950C,
respectively.
3. Wait for a 2-3 minutes and add 1 ml diluted (1 ml saliva /10 ml distilled water)
saliva into each test tube.
4. Place 1 drop of iodine solution on 4 watch glasses and take 1 drop from each
test tube to observe if there is any starch left (Starch gives blue color with
iodine solution).
5. Record the time for the hydrolysis of 1 ml starch in each test tube.

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DATA SHEET
Experiment No. 4
Hydrolysis of Starch By α - Amylase

Name: _____________________________ Rating: ____________________

Section:____________________________ Instructor: _________________
Schedule: __________________________ Date: _____________________

Results:

Test tube Time for the Hydrolysis

1
2
3
4

Questions:

1. What is hydrolysis?

2. What is the aim of using phosphate buffer in each starch solution?

3. Explain the significance of each temperature.

4. Calculate the rate of hydrolysis reaction in each test tube.

5. Compare your reaction rate results at each temperature. Explain briefly

Conclusion(s):

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Experiment No. 5
Preparation of Aspirin

Objective:
1. To learn how to prepare a useful ester - aspirin.

Materials/Chemicals:
Salicylic acid, acetic anhydride, phosphoric acid, ethanol, Erlenmeyer flask,
graduated cylinder, dropper, Buchner funnel, filter paper, beaker, hot plate or electric
stove, dropper and watch glass

Discussion:

Over history, many compounds obtained from nature have been used to cure ills or
to produce an effect in humans. These natural products have been obtained from
plants, minerals, and animals. In addition, various transformations of these and other
compounds have led to even more medically useful compounds. During this semester,
you will have an opportunity to isolate some pharmacologically active natural products
and to synthesize other active compounds from suitable starting materials.
Analgesics are compounds used to reduce pain, antipyretics are compounds used
to reduce fever. One popular drug that does both is aspirin. The Merck Index, which is
an encyclopedia of chemicals, drugs and biologicals, lists the following information
under aspirin: acetylsalicylic acid; monoclinic tablets or needle-like crystals; mp 135 °C
(rapid heating); is odorless, but in moist air it is gradually hydrolyzed into salicylic and
acetic acids; one gram dissolves in 300 mL of water at 25 °C, in 100 mL of water at 37
°C, in 5 mL alcohol, in 17 mL chloroform.

SYNTHESIS OF ASPIRIN (acetylsalicylic acid)

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Procedure:

1. Weigh 3 grams of salicylic acid and place it in an Erlenmeyer flask.

2. Add 6 ml of acetic anhydride into the Erlenmeyer flask containing salicylic acid.
Mix thoroughly.
3. After mixing, add 6 drops of phosphoric acid (H3PO4) and then shake it again.
4. Place the Erlenmeyer flask in warm water for about 15 minutes until the
solution becomes clear and transparent.
5. Add 20 drops of distilled cold water and then add another 20 ml of cold distilled
water to bring down the temperature slowly.
6. Cover the flask and place it in a refrigerator to undergo crystallization.
7. Filter the crystals using a Buchner funnel with a suction vacuum and washed
the crystals with water.
8. Place the crystal in a beaker and then add 10 ml of ethanol.
9. Place the beaker in warm water to dissolve the crystals with continuous
stirring.
10. Add 25 ml of warm water and seal or cover the beaker as this will recrystallize
as the solutions gets colder. To speed recrystallization, you can place it in a
refrigerator.
11. Once the aspirin is recrystallized, filter again as in procedure #7.
12. Transfer the crystals into a filter paper to complete the drying process.
13. Weigh and record the weight of aspirin produced

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DATA SHEET
Experiment No. 5
Preparation of Aspirin

Name: _____________________________ Rating: ____________________

Section:____________________________ Instructor: _________________
Schedule: __________________________ Date: _____________________

Results:

Weight of aspirin produced: ____________ grams

Questions:

1. What is the chemical name of aspirin?

2. Why did we recrystallize aspirin using ethanol alcohol?

3. Why is the aspirin washed with cold water?

4. Enumerate important uses of aspirin.

5. A polymeric material, which is a polyester, is formed in this reaction. Polyesters are

often made from dicarboxylic acids and diols. In this case, one molecule (salicylic acid)
provides both the "alcohol" and the carboxylic acid. Write a structure for the ester
formed from acetic acid and ethanol. If you have difficulty, you may research on this.

Conclusion(s):
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Experiment No. 6
Chemiluminescence

Objective:
1. To produce light with the aid of an oxidizing agent.

Materials/Chemicals:

ammonia water (NH3.H2O), potassium tricyanocuprate K2[Cu(CN)3], hydrogen

peroxide (H2O2). Erlenmeyer flask, graduated cylinder,
alternate chemical: light stick containing fluorescent liquid and diphenyl oxalate in
internal ampule

Discussion:

There are reactions where light is emitted without the emission of a considerable
amount of heat. This phenomenon is called cold light or chemiluminescence.
In this experiment, chemiluminescence accompanying the oxidation of luminol is
demonstrated. Hydrogen peroxide will oxidized a mixture of ammonia water and
potassium tricyanocuprate and energy will be emitted in the reaction in the form of
light quanta.
There are several consumer goods based on luminescence that are available in the
market. To name a few are glowing nail polishes, glowing sticks and glowing flowers.
Glowing nail polishes are quite popular. They shine stronger under UV light. Glow sticks
contain some fluorescent compound in a glass ampule surrounded by a solution of an
oxidizer.
Chemiluminescence can be observed in nature. For example, the green glow of the
common glow-worm is known. Luciferin is the dye present in the worm activated by
the enzyme luciferase and air oxygen.

Procedure:

1. Prepare 100 ml solution of ammonia water with potassium tricyanocuprate

(1:1 ratio).
2. Add 30 ml solution of hydrogen peroxide in a dark room in order to oxidize
luminol.
3. You may add enough hydrogen peroxide slowly to see the reaction in a dark
room.

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4. Add more hydrogen peroxide to produce light. See the effect
of hydrogen peroxide in the mixture.

Note: The room should be totally dark or cover any source of light in order to see
the reaction and production of light.

Alternate procedure:

1. Pour the fluorescent liquid inside the glow stick in a test tube and clamp the
test tube diagonally.
2. Slowly add drop by drop of hydrogen peroxide using a dropper into the test
tube by allowing the peroxide to slide on the side of the test tube. No lights
should be present.
3. Cover the beaker and a yellowish glow accompanied by the mixing of
reactants.

Note: Try using glowing stick and bent it to make bracelet or necklace and see
the effect on a dark room. When the stick is bent, the ampule breaks and the
fluorescent compound is oxidized thus emitting light. Different light colors are
obtained with different dyes.

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DATA SHEET
Experiment No. 6
Chemiluminescence

Name: _____________________________ Rating: ____________________

Section:____________________________ Instructor: _________________
Schedule: __________________________ Date: _____________________

Results:

What is your observation(s):

Questions:

1. What is the chemical formula of luminol?

2. What is the principle behind this experiment?

3. What color is usually produced by the following dyes?

a.) 9,10 - diphenylanthracene - ______________________________________

b.) 1 – chloro – 9,10 – bis (phenylethynyl)anthracene - ____________________

c.) Rhodamine – B - ________________________________________________

d.) 5,12 – bis (phenylethynyl)napthacene - ______________________________

4. Define Chemiluminescence.

Conclusion(s):
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Practical Experiment No. 7

Preparation of Liquid Dishwashing

Objective:
1. To be able to produce liquid dishwashing as an application of organic chemistry
knowledge and concepts.

Materials/Chemicals:

Distilled water (15 liters), sodium hydroxide or caustic soda, LABSA (linear alkyl
benzene sulfuric acid), coco diethanolamide (CDEA), colorant, essence or scent,
sodium chloride (technical grade), methyl paraben, ethylenediaminetetraacetic acid
(EDTA), platform balance, 1 liter beaker, wooden stirrer , empty plastic bottle
containers , wide mouth container with 15-20 liters capacity

Discussion:

Dishwashing liquid or known as dishwashing soap is a detergent used to assist in

dishwashing or removing food from used dishes or table ware. It is usually a highly
foaming mixture of surfactants of low skin irritation and is primarily used in for hand
washing of glasses, plates, cutlery and cooking utensils in a sink.
Your skin is semi-permeable and can be an entry point for toxins. If you're using a
toxic dish soap and are immersing your hands in it, sometimes several times a day,
you could be leaving yourself vulnerable to health problems.

Toxins found in non-organic dishwashing liquid include synthetic

fragrances, formaldehyde, silicone, acrylate, preservatives, genetically modified
organisms, and even foaming and coloring agents, just to name a few. Such
chemicals are not necessary for clean dishes and they're certainly not good for your
health or your skin, as some can cause immediate allergic reactions. Wearing gloves
might seem like an easy solution but these chemicals are still toxic to the environment
and don't really "go away" when they go down the drain.

Procedure:

1. Dissolve sodium hydroxide flakes in water.

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2. Slowly add LABSA with slow agitation until the solution
becomes clear for 10-15 minutes or as necessary.
3. Dissolve the methyl paraben in CDEA.
4. Dissolve EDTA and sodium chloride separately with water.
5. Add water to LABSA mixture.

6. Add CDEA solution, EDTA solution followed by sodium chloride solution and
mixed well.
7. Add colorant and essence.
8. Set aside to allow clearing of solution.
9. Pack in plastic containers with labels.
10. Submit your finished product by group to your instructor with complete
packaging and labeling.

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DATA SHEET
Practical Experiment No. 7
Preparation of Liquid Dishwashing

Name: _____________________________ Rating: ____________________

Section:____________________________ Instructor: _________________
Schedule: __________________________ Date: _____________________

Results:

State your observation(s):

Questions:

1. What are the organic compounds used in the preparation of liquid dishwashing?

2. What are your learning insights in performing this experiment?

Conclusion(s):

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Practical Experiment No. 8

Preparation of Perfume (Eau De Parfum)

Objective:
1. To be able to produce perfume as an application of organic chemistry
knowledge and concepts.

Materials/Chemicals:

Fragrance oil (30 ml), fixative oil, dipropyleneglycol (DPG), perfume grade
alcohol or deodorized alcohol, distilled water, empty perfume glass containers,
empty ambered bottle (250 ml), Erlenmeyer flask, beaker, graduated cylinder,
stirring rod

Discussion:
Perfume is a classic gift, but it's even better if the perfume you give is a scent that
you created yourself, especially if you package it in a beautiful bottle. Perfume you
make yourself is free from synthetic chemicals and is fully customized to your personal
taste.

Eau de parfum (EDP) is usually 15-20% oil based concentration. It is a strong

concentration for perfume and usually it is a long lasting perfume. If the perfume is too
strong, you can dilute it with more water. If you want your perfume to retain its scent
longer, add a tablespoon of glycerin to the perfume mixture.

The essential oils that you use form the basis of your perfume. These essential
oils are called the 'notes' of the perfume. The base notes are the part of the perfume
that lasts the longest on your skin. The middle notes evaporate a little more quickly.
The top notes are the most volatile and disperse first. Bridge notes have intermediate
evaporation rates and serve to tie a scent together. Sometimes other substances are
added to a perfume, such as sea salt (ocean scent), black pepper (spicy), camphor,
and vetiver. Since the essential oils evaporate at different rates, the way a perfume
smells changes over time as you wear it. Here are some examples of common base,
middle, top, and bridge notes.

Procedure:
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1. Put the 30 ml fragrance oil (depending on your choice) into your beaker.
2. Add your 10 ml fixative. Fixative is just like a preservative that maintains the
freshness of the perfume. It locks the scent of the perfume to have a long lasting
fragrance.
3. Mix well using a stirrer.
4. Add 5 ml dipropyleneglycol and mix well for about 2-3 minutes. DPG is a solvent

commonly used in cosmetic products and incense. It controls the fragrances of

the perfume to become safe to skin so as not to burn the skin.
5. Add 5 ml of distilled water and mix it again.
6. Add 100 ml of perfume grade alcohol or deodorized alcohol and mix it well.
7. Transfer it to an ambered bottle container and cover it tightly.
8. Store it in a refrigerator for 7 days or 14 days in a cabinet at room temperature not
directly affected by the light of the sun.
9. After the ageing process, transfer your perfume in your desired glass container
and label it.
10. Have it check by your instructor.

Note: If you add 100 ml alcohol then you will produce a 20% oil based perfume
which is very strong perfume. If you want to reduce the concentration to 15% oil
based, you may add another 50 ml of your alcohol.

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DATA SHEET
Practical Experiment No. 8
Preparation of Perfume

Name: _____________________________ Rating: ____________________

Section:____________________________ Instructor: _________________
Schedule: __________________________ Date: _____________________

Results:

State your observation(s):

Questions:

1. What are the organic compounds used in the preparation of perfume?

2. What are your learning insights in performing this experiment?

Conclusion(s):

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BIBLIOGRAPHY

A. Books and Journals

Helmenstine, Anne Marie PhD., Biomedical Sciences, Perfume making in

college students, Chicago TechnicalCollege, January 5, 2019

James Chickos, David Garin, and Valerian D'Souza. Organic Chemistry

Laboratory Manual, University of Missouri – St. Louis; Chemistry), USA October 2017

McMurry, John, Organic Chemistry, a biological approach. 2nd edition Andover

Cengage Learning, USA 2015

Oxtoby, D.W. et. al., Principles of Modern Chemistry; Saunders College

Publishing, 4th Edition (2018), p. 600 - 614

Poon, Thomas, Joint Science Department, Claremont Mckenna College, CA,

USA. 2017

Sigmon, Jane. Hydrolysis of Starch, American Society for Microbiology, USA

2018

Zare, Richard. et. al. Laser Experiments for Beginners; 2016, University
Science Books, Sausalito, California.

B. Internet Sources

http://onelearningsolution.blogspot.com/2015/01/212-laboratory-activity-determining.htm
l

http://jan.ucc.nau.edu/~jkn/235L2-Distillation&GC.htm

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