International University Ho Chi Minh City – Vietnam National University

Department of Applied Chemistry

ORGANIC CHEMISTRY LAB MANUAL

Prepared by Dr. Hoang Le Son

Revised by Dr. Le Quang Phong
and Mr. Nguyen Thanh Phong

SEMESTER I 2022-2023
Giảng viên phụ trách:

Dr. Lê Quang Phong

LABORATORY SAFETY GUIDELINES
If you do not have time to do things correctly and safely, with adequate time for thought, please stay
away from the laboratory.

A. CLOTHING
1. WEAR APPROVED EYE PROTECTION AT ALL TIMES. Very minor laboratory accidents, such
as the splattering of solution can cause permanent eye damage. Wearing laboratory goggles can prevent
this eye damage. In the chemistry teaching laboratories safety glasses (goggles) of an approved type must
be worn by all persons in the room at all times that anyone is working with or transporting glassware or
conducting any experimental work. Experimental work includes simple tasks such as transporting
chemicals or glassware, obtaining quantitative measurements that involve non-sealed containers, etc.
Lightweight ‘‘visitors’ shields’’ or prescription glasses with side shields are acceptable only for laboratory
visitors if your institution permits them but are not suitable for routine laboratory work.

2. WEAR PROPER PROTECTIVE CLOTHING. Proper protective clothing must be worn by all
persons in the room at all times that anyone is working with or transporting glassware or conducting any
experimental work. Exposed skin is particularly susceptible to injury by splattering of hot, caustic, or
flammable materials. Students and instructors need to be protected from their necks to below their knees.
This requirement includes no shorts, no short skirts, no sleeveless garments, and no bare midriffs. Long lab
coats or aprons are required if shorts or short skirts are worn. Makeshift coverage such as shirts being used
as aprons, paper taped over the knees, etc., is not considered to be suitable. Tight fitting clothing, long
unrestrained hair, clothing that contains excessive fringe or even overly loose-fitting clothing may be ruled
to be unsafe. Long hair must be tied back and no dangling jewelry.

3. WEAR PROPER PROTECTIVE FOOTWEAR No sandals/flip-flops, no open-toed shoes, and no

foot covering with absorbent soles are allowed. Any foot protection that exposes any part of one’s toes is
unsuitable for wear in the laboratory.

B. HANDLING CHEMICALS
4. NEVER WORK IN A CHEMICAL LABORATORY WITHOUT PROPER SUPERVISION. Your
best protection against accidents is the presence of a trained, conscientious supervisor, who is watching for
potentially dangerous situations and who is capable of properly handling an emergency

5. NEVER INHALE GASES OR VAPORS UNLESS DIRECTED TO DO SO. If you must sample the
odor of a gas or vapor, use your hand to waft a small sample toward your nose.

6. EXERCISE PROPER CARE IN HEATING OR MIXING CHEMICALS. Be sure of the safety

aspects of every situation in advance. For example, never heat a liquid in a test tube that is pointed toward
you or another student. Never pour water into a concentrated acid. Proper dilution technique requires that
the concentrated reagent be slowly poured into water while you stir to avoid localized overheating.
7. NEVER EAT, DRINK, OR SMOKE IN A CHEMICAL LABORATORY. Tiny amounts of some
chemicals may cause toxic reactions. Many solvents are easily ignited. Food and drinks are never allowed
in the labs. This includes all visible insulated water bottles or mugs, containers of water or flavored drinks,
containers of ice intended for consumption, etc. If a food or drink container is empty or unopened, it needs
to be inside a backpack, etc., and out of sight.

8. NEVER RETURN RESIDUE CHEMICALS TO THEIR ORIGINAL CONTAINERS. Used

chemicals might contaminate original chemical, thus never return them to their original containers.

C. HANDLING EQUIPMENTS
9. BE CAREFUL WITH GLASS EQUIPMENT. Cut, break, or fire-polish glass only by approved
procedures. If a glass-inserter tool is not available, use the following procedure to insert a glass rod or tube
through a rubber or cork stopper. Lubricate the glass and the stopper, protect your hands with a portion of
a lab coat or a towel, and use a gentle twisting motion to insert the glass tube or rod.

10. NEVER PIPET BY MOUTH. Always use a mechanical suction device for filling pipets. Reagents
may be more caustic or toxic than you expect.

11. USING INSTRUMENTS. Before using any instruments in the lab, read the manual carefully. Ask the
instructor or lab technician if needed.

D. BEHAVIORS
12. NEVER PERFORM AN UNAUTHORIZED EXPERIMENT. ‘‘Simple’’ chemicals may produce
undesired results when mixed. Any experimentation not requested by the laboratory manual or approved
by your instructor may be considered to be unauthorized experimentation.

13. NO REMOVAL OF CHEMICALS OR EQUIPMENT FROM THE LABORATORY. The

removal of chemicals and/or equipment from the laboratory is strictly prohibited and is grounds for severe
disciplinary action.

14. NO HORSEPLAY. Horseplay and pranks do not have a place in instructional chemistry laboratories.

15. NO LAPTOPS, TABLETS, ETC. Laptops, tablets (iPad, tab ...) are not allowed in the lab. Only
notebook and pen/pencils are allowed.

16. CLEANING. Wash hands with soap and water after performing all experiments. Clean (with detergent
powder), rinse, and dry all work surfaces and equipment at the end of the experiment.
E. EMERGENCY PROCEDURES
17. KNOW THE LOCATION AND USE OF EMERGENCY EQUIPMENT. Find out where the safety
showers, eyewash spray, and fire extinguishers are located. If you are not familiar with the use of emergency
equipment, ask your instructor for a lesson.

18. DON’T UNDER-REACT. Any contact of a chemical with any part of your body may be hazardous.
Particularly vulnerable are your eyes and the skin around them. In case of contact with a chemical reagent,
wash the affected area immediately and thoroughly with water and notify your instructor. In case of a
splatter of chemical over a large area of your body, don’t hesitate to use the safety shower. Don’t hesitate
to call for help in an emergency.

19. DON’T OVER-REACT. In the event of a fire, don’t panic. Small, contained fires are usually best
smothered with a pad or damp towel. If you are involved in a fire or serious accident, don’t panic. Remove
yourself from the danger zone. Alert others of the danger. Ask for help immediately and keep calm. Quick
and thorough dousing under the safety shower often can minimize the damage. Be prepared to help, calmly
and efficiently, someone else involved in an accident, but don’t get in the way of your instructor when he
or she is answering an emergency call.

These precautions and procedures are not all you should know and practice in the area of laboratory safety.
The best insurance against accidents in the laboratory is thorough familiarity and understanding of what
you’re doing. Read experimental procedures before coming to the laboratory, take special note of potential
hazards and pay particular attention to advice about safety.

Take the time to find out all the safety regulations for your particular course and follow them meticulously.
Remember that unsafe laboratory practices endanger you and your neighbors. If you have any questions
regarding safety or emergency procedures, discuss them with your instructor. Then sign and hand in the
following safety agreement.
 Specific Guideline for Organic Chemistry Lab
1. Class starts at 13:15. Arriving after 13:35 will be marked as “Absent” for the whole lab.
2. Missing any lab will result in automatic FAIL.
3. This course includes 12 sessions, in which we will have 1 orientation day, 1 midterm
day, 1 final exam day and 9 lab days.
4. Regarding the group report:
a. The report is due 1 week after the corresponding lab day. You are required to submit
the soft copy on Blackboard. In case of technical issue, please email your report to
TA and instructor before the deadline.
b. Late submission is not accepted.
c. Overall format:
● No hand writing, only typing.
● Include group number, section number and names of group members.
● Pictures/figures must be labeled and captioned. Pictures have to demonstrate the
text well (no blurry, no out-focused, using white background). Do not just include
pictures with no explanation.
● Use 1.15 spacing for the tables, and 1.5 for the text. Font Times New
Roman/Calibri, size 12 with margin set to “Normal” and “Justify” paragraph.
d. The report should have the format resembling that of a scientific article, which means
it includes
● Abstract
● Introduction
● Materials and Methods
● Results and Discussion
● Conclusion
● References (ACS Style).
e. For each experiment, the report should be to-the-point and fit into 4-6 pages,
including tables and figures. Longer report is not advised.
f. If plagiarism is detected, depending on the originality analysis, your group will get
20-100% point deduction.
TABLE OF CONTENT
EXPERIMENT ONE – Determination of melting point
EXPERIMENT TWO – Recrystallization

EXPERIMENT THREE – Thin layer chromatography

EXPERIMENT FOUR – Column chromatography
EXPERIMENT FIVE – Simple distillation
EXPERIMENT SIX – Reflux reaction
EXPERIMENT SEVEN – Liquid-liquid extraction
EXPERIMENT EIGHT – Fischer ester synthesis
EXPERIMENT NINE – Solvent Effects in an SN1 Solvolysis Reaction
 Experiment #1: MELTING POINT DETERMINATION
I. PURPOSE
To introduce the technique of melting point determination.
II. INTRODUCTION
The melting point of a solid can be easily and accurately determined with only small
amounts of material and, in combination with other measurements, can provide rapid
confirmation of identity. The most accurate method of determination is to record a cooling curve
of temperature versus time. However, this approach requires quite large amounts of material and
has been exclusively replaced by the capillary method.
Capillary Melting Point Determination
The method involves placing a little of the sample in the bottom of a narrow capillary tube
that has been sealed at one end. The determination is made using a Melting Point Apparatus that
simultaneously heats both the sample tube and a thermometer. The temperature range, over
which the sample is observed to melt, is recorded. Some pure materials have a very narrow
melting range, perhaps as little as 0.5–1.0 °C, while more typically a 2–3 °C range is observed.
Data is typically recorded as, for example, mp 232–234 °C. Though formally denoting the
melting range, this piece of data is almost universally referred to as a melting point (mp).
The rate of heating, controlled by a dial, should be kept relatively low, especially for
samples with a low melting point, to ensure that the thermometer reading represents, as
accurately as possible, the true temperature experienced by the sample tube (since the transfer of
heat within the apparatus is relatively slow).
Although a pure solid might be expected to have a single, sharp melting point, most
samples are observed to melt over a narrow range of a few degrees Celsius. The observation of a
melting range may be a result of the heating process involved in capillary measurements
(mentioned above), may reveal the presence of inhomogeneities in the macroscopic nature of the
solid sample, or may indicate the presence of other substances in the sample (contaminants or
by–products of the method used to prepare the materials).
Melting Points as Criteria of Purity
Thermodynamics tells us that the freezing point of a pure material decreases as the amount
of an impurity is increased. The presence of an impurity in a sample will both lower the observed
melting point, and cause melting to occur over a broader range of temperatures. Generally, a
melting temperature range of 0.5–1.0 °C is indicative of a relatively high level of purity. It
follows that for a material whose identity is known an estimate of the degree of purity can be
made by comparing melting characteristics with those of a pure sample.
Melting Points as a Means of Identification and Characterization
For pure samples a clear difference of melting point between two materials reveals that
they must possess different arrangements of atoms, or configurations. If two materials are found
to have the same melting point then they may, but not necessarily, have the same structure.
Clearly, the recording of a melting point is a desirable check of purity and identity but must be
combined with measurements from other analytical techniques in order to unambiguously
identify a material and assess the purity.
Mixed Melting Points
 Mixtures of different substances generally melt over a range of temperatures that concludes
at a point below the melting point of either of the pure components—each component acts as an
impurity in the other. Two pure substances, with sharp melting points, can be shown to be
different by mixing them and recording the lowered melting point range. This type of
experiment provides a means by which to confirm a proposed identity for an unknown sample: if
a sharp melting point is observed for a mixture of the unknown with a genuine sample then it is
highly likely that the samples are identical.
Melting Points and Molecular Structure
Melting points are notoriously difficult to predict with any accuracy or confidence.
Systematic variations of melting point with variation in structure are not as obvious or
predictable as with boiling points. However, the sometimes surprising variations are often only
highlighted in very closely related molecules and the obvious general rule can be applied:
melting points do generally increase with increasing molecular weight. The difficulties involved
in predicting melting points are a result of the problems associated with predicting molecular
packing in crystals. Many, potentially conflicting, factors play a role in determining melting
points including molecular shape, interactions between groups within the molecule, and the
degrees of freedom the molecule possesses within the crystal.
III. PROCEDURE
For the melting point determinations you will use the melting point instruments housed in
the laboratory. There are three experiments for your group to perform.
The capillary melting point tubes should be filled by crushing the sample to a fine powder
on a watch glass with the end of a glass rod, and then introducing this powder to the tube by
pressing the open end into the powdered sample.
Once a 2–3 mm depth of powder has been introduced the tube should be inverted and the
sealed end tapped on a hard surface until the sample rests at the bottom.
Part 1: Determine the melting points of pure substances
Obtain 0.2 g of urea and 0.2 g of cinnamic acid. The melting point of these are determined
individually via described method. For both samples the range should be very close to 131–133
°C. Record your values.
Part 2: Determine the melting points of compounds mixture
Mixtures of urea and cinnamic acid in molar percentage ratios of 100:0, 90:10, 80:20,
60:40, 50:50, 40:60, 20:80, 10:90 and 0:100 ratios should be prepared by a volunteer. Carefully
record the melting point for your mixed sample. Once all the data has been collected plot both
the beginning of melting and complete melting temperatures versus the molar percentage ratios
of urea and cinnamic acid.
Part 3: Identify an unknown substance by melting point
Determine the melting point of an unknown sample provided by your TA or instructor
(reminder: a crude melting point run will be necessary to determine the approximate melting
temperature). Confirm the proposed identity of your unknown sample by performing a mixed
melting point determination with an authentic sample of the suspected material. Recall that the
melting point will not be lowered or broadened if the materials are identical. Your sample will be
one of the compounds listed in Table 1-1.
 Material Melting Point (˚C)
Acetanilide 113.5 – 114.0
Fluorene 114.0 – 116.0
Cyanoacetamide 121.0 – 122.0
Benzoic acid 121.5 – 122.0
Benzamide 128.0 – 130.0
Urea 131.0 – 134.0
Benzilic acid 150.0 – 153.0
Adipic acid 152.0 – 154.0
Salicylic acid 158.5 – 159.0
Sulfanilamide 165.0 – 166.0
Succinic acid 184.5 – 185.0
p–Terphenyl 210.0 – 211.0

Table 1-1. Melting point data for the possible unknown samples.
IV. CLEAN UP
Put all used tubes in the solid waste container marked for this purpose.
V. REPORT
Record all of the melting point for the three parts of the experiment in the results section of
the report. In the discussion section describe what is revealed by each measurement about the
sample in question.
VI. QUESTIONS FOR DISCUSSION
1. Why should samples for melting point determination be finely powdered?
2. Why is it important that the heating rate be carefully controlled once near the suspected
melting point of a sample?
3. Describe and analyze the trend lines for the initial and final melting temperatures versus
composition revealed by the graph you generated in Part 2 of this experiment. To what extent
do you think it would be possible to employ this graph to determine the composition of an
unknown urea and cinnamic acid sample?
 Experiment #2: RECRYSTALLIZATION
I. PURPOSE
To introduce the technique of recrystallization through the recrystallization of crude benzoic
acid. To recrystallize an unknown compound and determine the identity via melting point
determination.
II. INTRODUCTION
Purification of a solid by recrystallization from a solvent relies on the fact that different
substances are soluble to differing extents in a given solvent. In the simplest case all of the
impurities present in a solid sample will be so much more insoluble in the chosen solvent that all
that remains in solution is the pure dissolved product (the solute). The process of
recrystallization can be broken down into a number of discrete steps:
1. Choosing the solvent
An essential characteristic of a successful solvent is that the compound be soluble in the hot
solvent but insoluble when the solvent is cold. The impurities are either insoluble in the solvent
at all temperatures or remain at least moderately soluble in the cold solvent.
In order to select a solvent system, tests can be performed with small amounts of material in
test tubes: a few drops of solvent are added and if the material proves insoluble then the tube is
heated to see if the material will dissolve at a higher temperature — if so, then a good solvent for
recrystallization of that material may have been identified. If the material appears insoluble at
high temperature a little more solvent can be added, drop by drop, to ascertain whether
dissolution of the solute will occur in a larger volume of solvent.
A solvent should be rejected if: the material appears readily soluble in the cold solvent, is not
soluble to any appreciable extent in the hot solvent even when the volume of solvent is
increased, or requires an impractically large volume in order to fully dissolve the crystals.
2. Dissolving the sample
An Erlenmeyer (conical) flask should be used of such a size that it is only filled to around
half–way when all the solvent has been added. The solid sample is introduced together with
about 75 % of the amount of solvent might be required. Using less solvent at this stage is
advisable.
The flask is heated on a hotplate until the solute has dissolved, additional solvent can be
added to the hot solution as necessary to ensure complete dissolution. By a process of gradual
addition of solvent to the flask, until all the material has dissolved, you will ensure that the hot
solution is saturated and will deposit crystalline material once it is cooled.
3. Hot filtration
If some insoluble impurities remain suspended in solution it is necessary to filter the solution
while it is still hot, see Figure 2-1. The funnel, equipped with filter paper, is placed above the
flask, using a ring clamp, and the vapor from the warm solvent is allowed to warm the funnel.
Care should be taken to allow some space between the filter funnel and the flask to avoid a build
up of pressure in the flask. The hot solution is gravity filtered through the warm funnel and a
watch glass may be placed on top of the filter funnel to prevent heat loss during filtration.
 Figure 2-1. Hot filtration set up.
4. Cooling
Cooling the filtered solution will allow crystals to form. The rate of cooling has a role in
determining the size of the crystals: fast cooling will tend to generate more crystals of relatively
small dimensions, slow cooling might allow larger crystals to form. Usually the best compromise
of speed, convenience, and crystal quality, is simply to let the solution cool to room temperature
on the bench. To ensure maximum recovery of material the solution should be cooled in an ice–
water bath after the solution has been allowed to cool to room temperature. In some particularly
stubborn cases, crystal formation can be encouraged by providing a nucleation point, from which
the crystals may grow. This can be achieved by scratching the inner wall of the flask with a glass
stirring rod (or spatula), or if available a small amount of pure crystals can be added to the flask.
5. Cold filtration
When the crystallization process is judged to be complete the crystals need to be collected by
suction filtration (Figure 2-2). Both the funnel and suction flask should be chosen so that neither
will become more than half full during the filtration process. It is preferable that all of the
crystalline material is transferred to the funnel as a suspension in the crystallization solvent,
however it is sometimes hard to get all of the crystals moving freely by swirling the flask and
occasionally it will be necessary to add more ice–cold solvent in order to transfer the last of the
crystalline material.
 Figure 2-2. Typical set up for suction, or vacuum filtration.
6. Washing the crystals
Once the suction filtration process is complete the collected crystals should be washed with
a little more ice–cold solvent to remove final soluble impurities which would otherwise be left
on the surface of the crystals. The solvent used for this final washing should be as cold as
possible to minimize losses from the crystals redissolving.
7. Drying the crystals
Once the crystals have been collected on the suction funnel they can usually be dried by
continuing to draw air over them for a few minutes. The almost dry crystals should then be
spread on a filter paper to allow the last traces of volatile solvent to evaporate.
III. MATERIALS AND EQUIPMENTS
− Vacuum filter − Ice pack
− Buchner funnel − Methanol
− 1 Filter flask − Ethanol
− 1Erlenmeyer flask 125 mL − Distilled water
− 1 Cylinder 50 mL
− 1 Spatula
− 1 Stirring rod
− Hot plate
− Balance
− Water bath
− 3 50ml beakers
IV. PROCEDURE
Part 1: Recrystallization of Benzoic acid
1. Using a weighing paper, weigh out about 0.5 g of impure Benzoic acid and transfer
it to a 125-ml Erlenmeyer flask. Add about 20 ml distilled water, using a graduated
cylinder, to the flask and bring the mixture to the boiling point by heating on a hot
plate, while stirring the mixture and boiling gently to dissolve benzoic acid
completely.
 2. Remove the flask from the hot plate and examine the solution. If there are particles
of benzoic acid still undissolved, then add an additional amount of hot or cold water
in small increments and resume heating the solution. Keep adding water in small
amounts (several drops at a time from a Pasteur pipette) until all of the benzoic acid
is dissolved and the solution is boiling.
3. When the solution is completely clear (though not necessarily colorless) and no
solid benzoic acid is visible, then add additional 10-15 ml water to the mixture and
place the Erlenmeyer flask on a countertop where it will not be disturbed and cover
with an upside-down small beaker (to prevent dust contamination).
4. Allowing the flask to cool slowly or put it into an ice bath and note the
development of crystals in the first 5-10 minutes. If crystallization does not occur
after 10 minutes, scrape the sides of the flask above the level of the solution with
the sharp end of a glass rod hard enough to audibly scratch the interior surface of
the flask to help inducing crystallization.
5. When the crystals have formed completely, collect your solid chemical by filtration
on a in a Büchner funnel apparatus. Get all the crystals out of the flask using a
spatula or stirring rod. Rinsing with 1 or 2 ml of cold water helps get the crystals
out of the flask, and rinsing helps remove impurities.
6. Use a spatula to lift the filter paper and crystals out of the Buchner funnel, then
allow them to dry completely, and transfer the dry sample to a pre-weigh weighing
boat. Determine the of recovered benzoic acid crystal.
7. Calculate the percent recovered using the following written formula and determine
the melting point of your recrystallized benzoic acid.

% Recovered = Weight of benzoic acid obtained after recrystallization /

Weight of benzoic acid before recrystallization x 100

Part 2: Recrystallization and Identification of an Unknown Compound

1. Obtain a sample of an unknown compound from you Instructor or TA and record
its identification number in your notebook. All of the unknowns will crystallize
from one, or more, of the following solvents: water, methanol, or ethanol. The
unknown will be one of the materials listed in Table 2-1.
2. To identify the solvent for recrystallization, procure 0.5 g of the unknown sample
and observe the solubility of the sample in those solvents at room temperature and
high temperature. Carefully retrieve the crystal and identify the substance based on
melting point of the dried product. Recrystallize again in a beaker.

Material Melting Point (˚C)

Acetylsalicylic Acid 134–136
Adipic Acid 152–153
Benzamide 128–129
Benzil 94–95
Benzilic Acid 150–153
Benzoic acid 122–123
Cholesterol 147–149
 2–Nitrobenzoic Acid 140-142
Salicylamide 140–144
Salicylic Acid 158–160
Sulfanilamide 164–166
trans–Stilbene 122–124

Table 2-1. Melting point data for the possible unknown samples.
CLEAN UP
All solvent wastes from vacuum filtrations should be placed in the non–chlorinated waste
container. All of your solid samples should be placed in the solid organic waste container. Wash
all of your glassware and return to your drawer.
REPORT
In the observations/data section of your report record the solvent types you chose, the
method of adding solvents, the method of cooling down solutions, the dried weight, percentage
of recovered material, and melting points, for the two parts of the experiment. Also include
descriptions of both the crude and recrystallized crystals. For part 2, record the steps you took to
select the solvent for recrystallization of the unknown.
QUESTIONS FOR DISCUSSION
1. What properties should ideally be possessed by a recrystallization solvent?
2. Why are crystals washed with additional solvent when collected? Why cold solvent?
 Experiment #3: THIN LAYER CHROMATOGRAPHY
I. PURPOSE
To introduce chromatography techniques and to learn how to manipulate these techniques
to both separate mixtures of compounds and to analyze materials.
II. INTRODUCTION
Thin Layer Chromatography (TLC) is a solid-liquid technique in which the two phases are a
solid, stationary phase and a liquid, mobile phase. The solid phase you will be using in today’s
lab is a plastic plate covered with an adsorbent, in this case, silica gel. Aluminum is another
common solid phase used. Because silica is a polar molecule, the components of the solution we
will use in lab today will be separated based on their relative polarities. The more polar a
molecule, the higher affinity it will have for the more polar silica plate and will therefore spend
less time in the mobile phase. As a result, it will move up the plate more slowly. Conversely, a
less polar molecule will spend more time in the mobile phase and will therefore move up the
plate more quickly. The speed at which the molecules will move up the plate thus depends on the
relative difference in polarity between the stationary and mobile phases, and will vary depending
on the nature of the stationary and mobile phases used for separation.
The following are some common uses of Thin-Layer Chromatography:

1. To determine the number of components in a mixture.

2. To determine the identity of two substances.
3. To monitor the progress of a reaction

The difference each molecule travels along the adsorbent in relation to how far the mobile phase
has traveled is called the Retention Factor (Rf) and can be used to identify molecules, as the
value is molecule specific. The Rf for any given molecule will vary depending on the mobile and
stationary phases used.
𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑒𝑑 𝑏𝑦 𝑠𝑢𝑏𝑠𝑡𝑎𝑛𝑐𝑒
𝑅𝑓 = 𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑒𝑑 𝑏𝑦 𝑠𝑜𝑙𝑣𝑒𝑛𝑡 𝑓𝑟𝑜𝑛𝑡
 Figure 3-1. Example TLC chamber (left panel) and TLC plate (right panel). The distances
traveled by each spot and by the solvent front are shown along with the Rf value for each spot.

Note: TLC plates are pretty small so measurements with a standard ruler can only provide
Rf values to two decimal places.
The practice of TLC can be summarized in three discrete steps:
1. Sample Application
A small amount of sample is dissolved in a small volume of a solvent in which it is readily
soluble to provide a spotting solution. A glass spotter is dipped in this solution and then applied
to the plate to form a small concentrated spot at a marked position around 1 cm from the bottom
of a TLC plate.
The spot is made as small as possible to ensure, in as far as this is possible, that all of the
applied material starts the journey up the TLC plate from the same point. The amount of material
deposited on the plate is also very important: if the sample spot is too concentrated then the
adsorbent can become overloaded and the features in the TLC will streak and spread. If the spot
is rather weak it may be hard to see the features on the plate after it has been developed.
Preparing a good TLC plate requires practice and experience.
2. Development
The container used for development is most often a simple beaker covered with a watch
glass. Sometimes a piece of filter paper is placed along the wall of the beaker to ensure saturation
of the atmosphere with solvent vapor. Once a suitable solvent system for plate development has
been found, a few mLs are added to the beaker to provide a depth of a few millimeters. The plate
is placed in the beaker so that the bottom edge is immersed in the puddle of solvent. The solvent
level must be ensured to stay below that of the spot. A watch glass, which serves as a crude lid,
is placed on top. The solvent is allowed to run up the plate until the solvent front has reached
around 1 cm from the top edge of the plate. At this point the plate should be removed and the
furthest extent of the solvent front marked.
 3. Visualization
The solvent is allowed to evaporate from the adsorbent on the plate. If the substances under
investigation are colored, i.e. they absorb light in the visible region of the spectrum, then both
during and after development of the plate you will be able to see where the spots are and how
they have moved. If the components of the sample are not colored, an Ultra–violet (UV) lamp
may reveal the location of the spots and their positions may then be marked with a pencil.
Alternatively, the staining of a TLC plate with iodine vapor is among the oldest methods for the
visualization of organic compounds. There are also chemical methods for revealing the positions
of materials on plates that cannot be located by eye, UV lamp, or the iodine staining technique.

III. MATERIALS AND EQUIPMENTS

− Capillary − Silica plate for TLC
− Forceps − Methanol
− Mortar and pestle − Hexane
− 3 Vials − Chloroform
− 1 Cylinder 5 mL − Petroleum ether
− 1 Spatula − Acetone
− 2 Glass pipets − Boric acid
− 4 Beaker 100 mL − Oxalic acid
− Alcohol lamp − Iodine
− Aluminum foil
− UV chamber

IV. PROCEDURE
Part 1. TLC analysis of spinach leaf extract


1. Preparation of spinach leaf extract

1. Weigh out 5 grams of spinach leaf.
2. Tear the leaf into small (3-5 mm) pieces, and place them into a mortar with about
10 ml methanol.
3. Grind the mixture slowly and firmly with the pestle until the solvent is dark green.
Condense the extract.
4. Transfer as much of the liquid extract as possible into a glass vial, avoiding
ground leaves and stems. Heat in the waterbath gently to condense the extract to
1/5 of the initial volume.
5. Add a small portion of 5 mL hexane to the sample. The upper layer is collected as
hexane extract. Carry out the extraction exhaustively until the color of added
hexane layer is colorless. Heat in the waterbath gently to condense the extract to
1/5 of the initial volume.
2. Analytical Chromatography of spinach extract
1. Prepare the TLC chamber
− Prepare 4 different chambers by adding about 5 mL of developing
solvent (100% chloroform, 90:10 chloroform: methanol, 8:2 petroleum
ether: acetone, 5:5 petroleum ether: acetone) into a clean, dry beaker.
− Cap the chamber closely so that the air inside becomes saturated with
solvents.
 2. Prepare TLC plate for development
− Draw one faint pencil line around 1 cm from the bottom edge of the
TLC plate and one around 0.5 cm from the top (the solvent front).
− Using a capillary loaded with the extract in the vial, gently make a small
spot of the extract on the plate. Each spot should be even in size and not
too large to prevent smears and disturbed streaks. Allow the spots to air
dry thoroughly.
− Develop your plates in TLC chamber until the solvent reaches the
solvent front.
3. Observation and detection of phytocomponents
1. Observe the plates in room light and in a UV chamber. Record your observation.
2. Spray the plate with a reagent consisting of 15ml 3% boric acid solution and 5ml
10% oxalic acid. Heat the TLC plate on a hot plate and record your observation.
Examine the plat again under the UV light. Record your observation.
3. Use an alternative method to visualize the developed TLC plate: place the plate in
a chamber containing iodine. Record your observation after 1-2 minutes.

CLEAN UP
All of the liquid samples should be poured into the halogenated solvent waste container.
The different solvents and solvent mixtures used for TLC should be poured into the appropriate
solvent waste container, either halogenated or non–halogenated. Dispose of all vials and spotters
in the glass waste container.
REPORT
In the results section, please include sketches or photos of your TLC results.. Compare the
TLC results with respect to solvents of extraction, solvents of TLC development, and detection
methods. To do this, select 1, 2, 3 or a range of compounds and calculate their Rf.
QUESTIONS FOR DISCUSSION
1. What is the purpose of using two different solvents for the extraction at the beginning
of the lab? How are the chromatograms obtained from using these mixtures different
from each other?
2. What is the purpose of using different methods of detection? Aside from pigments,
what are other components that could possibly be found in spinach leaf extract,
according to your chromatograms?
 Experiment #4: COLUMN CHROMATOGRAPHY
I. PURPOSE
To introduce chromatography techniques and to learn how to manipulate these techniques
to both separate mixtures of compounds and to analyze materials.
II. INTRODUCTION TO COLUMN CHROMATOGRAPHY (LC)
Column chromatography is one of the most common and useful ways to separate mixtures
of materials and isolate large amounts of the individual components in pure form. You will
isolate three ferrocene derivatives from a mixture and determine the effectiveness of this
technique in terms of the purity of the products.
In this experiment liquid chromatography will be employed to separate the components of
a mixture. Much like TLC, the stationary phase will be silica gel and the mobile phase will be a
solvent, or mixture of solvents. In contrast to TLC, where the components of a mixture were
separated from top to bottom using capillary forces to draw solvent onto the bottom of a TLC
plate, column chromatography is performed using gravity to draw solvent downward through a
glass tube containing the silica gel.
Column chromatography allows separation of relatively large amounts of a product
mixture into pure individual compounds, each of which can be isolated, characterized, and
perhaps used for other experiments or syntheses. This type of purification and isolation is crucial
to all synthetic organic chemistry and the modern pharmaceutical industry could simply not exist
without it. Various types of liquid chromatography are also widely used in separations and
preparations of biological macromolecules such as proteins, carbohydrates, and DNA.
In practice
To determine the best solvent for column chromatography, you could run a bunch of trial
columns, but that would be expensive, wasteful, and very time–consuming. Instead, you will test
several solvent mixtures by TLC, just as you did in the previous experiment. Since the stationary
phase, silica gel, is the same for both TLC and column chromatography, the separations observed
by TLC will accurately reflect the separations seen on the column.
Once you have determined which solvent, or solvent mixture, you will use in this
separation, you will set up your column by neatly packing it with silica and solvent. The mixed
sample will be added to the column and its components will move through the column at rates
determined by their polarities and that of the mobile phase (the solvent) that you have selected.
Once you have collected the separate “fractions”, you can confirm the identity of each one
by comparative TLC with an authentic sample.
III. MATERIALS AND EQUIPMENTS
− 5 Vials − Silica plate for TLC
− Capillary − Silica for column chromatography
− 1 Cylinder 5 mL − Ethanol
− 1 Weighing bottle − Distilled water
− 1 Spatula
− 1 Forceps
− 3 Glass pipet
− 1 Beaker 100 mL
− Cotton
 − Aluminum foil

IV. PROCEDURE
1. Prepare spinach leaf sample for column chromatography
1. Prepare the spinach juice and centrifuge at 5000 rpm for 5 mins.
2. Collect the spinach supernatant.
3. Mix the spinach supernatant with silica gel in a careful and thorough manner. The
amount of silica gel should be sufficient to make the mixture dry, coarse and light
in color.
2. Prepare the column
1. Use a piece of wire to add a plug of cotton to the bottom of the column. There
should be enough cotton that the sand and silica will not fall out of the column.
However, too much cotton or cotton packed too tightly will prevent the eluent
from dripping at an acceptable rate.
2. Clamp the column to a ring stand. Rinse the inner side with the initial solvent
(ethanol). Stop rinsing and leave the solution 1 cm higher than the silica layer in
the column.
3. Prepare slurry of silica in the initial solvent by pouring dry silica into a beaker of
eluent. The ratio between silica and the eluent is approximately 1:2.
CAUTION: keep the dry silica in your hood and be careful not to inhale the
lightweight substance.
4. Quickly but carefully pour the slurry into the column. Stir and pour immediately
to maximize the amount of silica that goes into the column instead of remaining
behind in the beaker.
5. Allow solvent to drip into a clean flask. NEVER ALLOW THE SOLVENT TO
RUN DRY (the solvent should always be 1 cm higher than the surface of the
silica layer). Tap on the side of the column with a rubber stopper or tubing to
help the silica settle uniformly.
6. Use a Pasteur pipet to rinse any silica that is sticking to the sides of the column.
Allow the silica to settle.
3. Run the column and identify the fractions
1. Drain eluent from the column until no solvent remains above the surface of the
sand.
2. Carefully transfer the silica gel mixed with ink into the column. Do not disturb the
silica gel layer that has been packed. A piece of cotton should be placed on top of
the loaded silica layer to prevent disturbance when eluent is added.
3. Once the sample is loaded, gently add fresh solvent mixture to the column and
start the separation. Add more solvent or change the ratio of components in the
solvent as needed, at all times ensuring THE SOLVENT DO NOT RUN DRY
(the solvent should always be 1 cm higher than the surface of the silica layer).
Try to run the column continuously without closing the stopcock.
4. Collect the fractions of different colors from the column.
5. Perform TLC analysis against pure samples of each material. The developing
solvent 70:30 hexane: acetone is recommended.
V. CLEAN UP
 All of the solvents and samples left over from the TLC runs and liquid chromatography
should be poured into the non–halogenated solvent waste container. Silica gel should go into the
solid waste container. Dispose of all spotters in the glass waste container. Vials should be
thoroughly rinsed with acetone, the washings discarded in the non–halogenated waste container
and left in the hood to dry for the next lab group.
VI. REPORT
In the report you should include sketches of all of your TLC plates, and label all features
with their Rf values and identities.
QUESTIONS FOR DISCUSSION
1. Explain the choice of solvent for the separation of pigments in spinach.
2. What should be kept in mind while performing column chromatography?
 Experiment #5: SIMPLE DISTILLATION
I. PURPOSE
To familiarize with the techniques of simple distillation, including the set up and operation
that rely on volatilization or evaporation and subsequent condensation of liquids for separation
and purification.
II. INTRODUCTION

The vaporization of a liquid, relocation, and condensation of the resulting vapor is the basis
for a method of purification called distillation. Organic liquids containing very small amounts of
high boiling impurities are easily purified by “simple” distillation, that is, an easy distillation
with relatively poor separation. As the liquid vaporizes and the vapor comes into contact with the
thermometer bulb, the temperature rises. The temperature stabilizes at the boiling point and most
of the liquid distills. The temperature drops when there is no liquid left in the distillation flask.

Figure 3 : The apparatus for a simple distillation.

Steps in a Simple Distillation
Before proceeding, check the water flow through the condenser and make sure that all
ground-glass joints are snug. Plug the heating mantle into a rheostat, then plug the rheostat into
the wall socket. If you use a burner, check the vicinity for flammable solvents. (Do not use a
burner when distilling a flammable compound)
Slowly heat the mixture in the distilling flask to a gentle boil. You will then see the reflux
level (the ring of condensate, or upper level of vapor condensing and running back into the flask)
rise up the walls of the flask to the thermometer and side arm. At this time, the temperature
reading on the thermometer will rise rapidly until it registers the initial boiling point, which
should be recorded.
The vapors and condensate will pass through the side arm and into the condenser, where
most of the vapor will condense to liquid, and will finally drip from the adapter into the receiving
flask. The proper rate of distillation is one drop of distillate every 1-2 seconds. This rate is
achieved by controlling the amount of heat supplied to the distillation flask. A too slow rate
means that not enough vapor is passing the thermometer to give an accurate boiling point. A too
rapid rate leads to a lack of separation of components and also to uncondensed vapor being
carried through the condenser and into the room. It is generally necessary to increase the amount
of heat applied to the distillation flask (by increasing the rheostat setting) during the course of a
distillation.
Volatile impurities are the first compounds to distil. This first fraction, called the fore-run,
is generally collected separately. When the temperature has risen to the desired level and has
been recorded, place a fresh receiver under the adapter to collect the main fraction. Each time
you change a receiver, note the temperature reading and record the boiling range of the fraction.
Impurities that are higher boiling than the desired material are general not distilled, but are
left in the distillation flask as the residue. If higher-boiling impurities are present in large
quantities, the temperature may rise from the desired level as the impurities begin to distil.
However, the temperature frequently drops after the main fractions have distilled. If the
temperature drops at the end of a distillation, the last temperature to record is the highest
temperature, before the drop occurred.
>>>>SAFETY NOTE: Never carry out a distillation to dryness, but always leave a small
amount of residue in the distillation flask. The presence of boiling residue will prevent the flask
from overheating and breaking and will also prevent the formation of pyrolytic tars (difficult to
wash out).
In the simple distillation of the mixture of hexane and toluene, you will collect the distillate
in eleven fractions and measure the refractive index of each. Using this information, you will
construct two graphs: (1) boiling point versus total volume of distillate; and (2) refractive index
versus total volume of distillate (see sample graphs below). When you do the fractional
distillation, you will be able to compare the graphs of each experiment to see how the two types
of distillation differ in their ability to separate mixtures.
 Figure 4 : Sample graphs for the simple distillation of a mixture of hexane and toluene.
III. MATERIALS AND EQUIPMENTS
− Distillation apparatus with − Hexane
thermometer, stand, heating mantle − Toluene
and water pump − Distilled water
− 1 Round-bottom flask 100 mL.
− 1 Water bath
− 2 Glass pipet
− 2 Cylinder 10 mL
− 10 Test tubes and rack
− Aluminum foil
− Refractometer
IV. PROCEDURE
1. Add 25 mL of hexane, 25 mL of toluene to a 100 mL round bottom flask, clamped
to a ring stand and supported by a water bath.
2. Assemble eleven clean, dry test tubes of the same size: add 5.0 mL of water to one
and set it in a test tube rack. The volume of liquid in this test tube is used to
estimate (by comparison) the volumes of the distillation fractions.
3. Plug the heating mantle into a rheostat, then plug the rheostat into the wall socket.
A setting of 4 should bring this particular mixture to a boil. Increase or decrease the
 voltage setting to achieve a steady boil that maintains a drip rate of distillate of
about 1 drop every 2 seconds.
4. Collect about 1 mL of the initial distillate and then continue by collecting 5-mL
fractions in the cylinder and immediately transfer to test tubes. Record the
temperature at the start and at the end of each fraction. Cover each test tube after
the fraction has been collected to prevent evaporation. The very first 1 mL
portion is very important! Collect it carefully and run its refractive index
immediately.
5. The distillation is complete when the distillation flask is almost empty and the
temperature starts to drop or fluctuate. (Because of hold-up, your last fraction will
not be 5 mL.) When the apparatus is cool, transfer the residue to a small graduated
cylinder and record the volume.
6. Measure the refractive index of each fraction, along with the refractive indexes of
the starting hexane and toluene.
REPORT
Record all data. Construct two graphs: one of boiling point versus mL distilled and the
other of refractive index versus mL distilled. Use the upper value of the boiling range for each
fraction as the boiling point in your graph.
QUESTION FOR DISCUSSION
When a solvent is used to extract a small amount of a high-boiling product from a reaction
mixture, it is common practice to first distil the solvent by simple distillation, transfer the residue
to a smaller flask, and isolate the product in a second distillation. Why not just continue the first
distillation to isolate the product?
 Experiment #6: REFLUX REACTION

I. PURPOSE

To introduce the techniques of reflux while isolating fatty acids and lipids from coconut.

II. INTRODUCTION
A system is in reflux when sufficient heat is provided to make the liquid form vapor, as the
vapor rises it is cooled to a liquid and allowed to return to the reaction pot. In this way heat is
provided to a system without the loss of the liquid components in the form of vapor. A condenser
is normally attached to the reaction flask to provide the cooling mechanism. Cold water is
allowed to run through the condenser, however, to achieve the desired cooling effect a constant
flow of water must be maintained. Therefore, water should enter the condenser at the bottom,
and exit at the top. Figure 6-1 shows the typical set up. Notice that the reaction flask is clamped
such that it could be raised out of the heating bath in the event that it became over heated.

Figure 6-1. Typical set up for reflux.

III. MATERIALS AND EQUIPMENTS

− Reflux apparatus with stand, heating − Flaked coconut

mantle and water pump − Petroleum ether
− 1 Round-bottom flask 250 mL. − Diethyl ether
− TLC: − Acetic acid
o Capillary − TLC plate
 o 1 Beaker 100 mL − Acid solution for TLC visualization
o 1 Forceps
o Lamp
o Aluminum foil

IV. PROCEDURE

Part A: Maceration with petroleum ether.

1. Place 15 g of flaked coconut in round-bottomed flask (250 mL), add 100 mL of

petroleum ether and heat under reflux for 1 h.
2. Cool down the mixture and filter coconut turnings from the solution on a funnel
with filter paper.
3. Concentrate obtained solution under reduced pressure and weigh using rotary
evaporator.

Part B: TLC of obtained product

4. Develop the obtained product on a silica plate in a chamber saturated with

petroleum ether: diethyl ether and glacial acetic acid (80:20:10).
5. Remove the plate and allow the solvent to evaporate. After drying use a hot-plate
and heat the TLC plate carefully.
6. In a summary report the amounts and physical characteristics of obtained coconut.
V. REPORT
Record your observations through the various stages of the experiment including the reflux step
and extraction step. Provide an accurate sketch of your TLC plate, making sure to indicate the
number, intensity, and color and all features that you observe.
 Experiment #7: LIQUID-LIQUID EXTRACTION

I. PURPOSE

To introduce the techniques of liquid-liquid extraction the isolation of caffeine in foodstuff

and tablets.

II. INTRODUCTION

Liquid–liquid extraction is a technique used to separate compounds by employing differences

in their solubility in various solvents. This technique is important because it is very commonly
used as a routine separation/purification step at the end of an organic reaction.
Almost always, one of the layers is aqueous and the other is an organic solvent with which the
water will not mix (immiscible), such that two layers are formed and can be cleanly separated.
Since polar inorganic material will generally be water soluble, and the much less polar products
of organic reactions will be soluble in an organic solvent, a clean separation of polar from non–
polar material can be achieved by separating these two liquid layers.
The separatory funnel is a glass vessel with a stoppered top and a stopcock, tapered
construction used for liquid-liquid extraction. The two-layer mixture in the funnel is vigorously
shaken to thoroughly mix the two layers. A period of ten to thirty seconds of mixing is sufficient
to establish equilibrium, returning the funnel to the stand for a few seconds usually results in
clean separation of the two solvent layers.

Figure 7-1. Typical liquid-liquid extraction set up.

If the density of the extracting solvent is higher than that of water then it will form the
lower layer, the opposite is true if the density is lower than that of water. In the former case, the
lower layer, in which the desired organic material is now dissolved, can be collected by
removing the stopper from the neck of the funnel and turning the stopcock so that this layer
drains into an Erlenmeyer flask. If the top layer is desired than the aqueous layer must first be
drained off before the organic layer is collected.
III. MATERIALS AND EQUIPMENTS
 − 1 Erlenmeyer flask 250 mL − Vivarin tablet or tea
− Heating plate − Methylene chloride
− Vacuum filter − Benzene
− 1 Beaker 100 mL − Petroleum ether
− 1 Separatory funnel 250 mL with
stand
− Balance
− 1 Cylinder 10 mL
− 1 Cylinder 50 mL

IV. PROCEDURE
1. Add 30g tea in 200 mL of water in a 250-mL Erlenmeyer flask. Then boil the mixture for
10 min to ensure solution of the caffeine.
2. Filter off the tea by using the baby towel. Save the liquid.
3. Centrifuge the solution at 5000 rpm for 5 mins. Collect the liquid.
4. Cool the extract to room temperature, transfer it to a 250 mL separatory funnel, and
extract the aqueous solution three times with 20-ml portions of methylene chloride. Do
not shake the layers vigorously; use a swirling motion so as not to form an emulsion.
5. Combine the methylene chloride extracts and evaporate the extract to dryness on the
steam bath (HOOD). Do not heat the residue any longer than necessary to evaporate the
solvent. The residue that remains after evaporation of the solvent is crude caffeine; some
mint smell will be evident. The caffeine may be purified by recrystallization as described
below.
PURIFICATION PROCEDURE
1. Transfer the crude caffeine to a clean 50-ml beaker, add 5 ml benzene, and heat on a hot
water bath to dissolve the caffeine.
2. Remove the beaker from the heat source, add 10 ml petroleum ether (boiling range 60-
90°C), and allow the caffeine to crystallize.
3. Collect the product by suction filtration, wash it with 1 mL petroleum ether, allow it to
air-dry, and determine its melting point.
V. LAB REPORT
Calculate the percent yield before and after recrystallization and the melting point of your
recrystallized caffeine. Compare your melting point to the actual melting point.
VI. QUESTIONS FOR DISCUSSION
Can this procedure be scaled up for industrial manufacturing? Elaborate your thought.

Experiment #8: FISCHER ESTER SYNTHESIS

I. Purpose
To revise methods in organic chemistry that have been instructed and introduce the lab scale
synthesis of esters while reviewing relevant definition, concepts and understandings.

II. Introduction
Esters refer to a group of organic or inorganic acids derivatives, specifically the products in
the reaction between those and various alcohols. Structurally, esters are compounds in which the
hydroxyl group of a carboxyl group is replaced by an alkoxy or aryloxy group. The presence of
esters is widely acknowledged as they are biologically and industrially critical: glycerides
derived from glycerol and fatty acids possess high caloric values being a type of lipid commonly
found in living bodies; many low-weighted esters have distinctive smells that are ascribed to the
fragrance in essential oils and pheromones and thus are availed as artificial flavorings and
odorants.
The synthesis of ester, or Fischer esterification, is the process of forming an ester by
refluxing a carboxylic acid and an alcohol in the presence of an acid catalyst. The alcohol is
generally used as solvent and so, is present in large excess. Many different acids can be used as
catalyzing agents, such as sulfuric acid and toxic acid. A tetrahedral carbonyl addition
intermediate created amidst the process of esterification tends to lose a water molecule to form
the ester. The reaction is generalized as below:

Numerous common esters with mimicking odors to common fruits such as orange (octyl acetate),
apple (methyl butanoate), grape (methyl 2-aminobenzoate), etc. can be synthesized and refined
in the laboratory. In this experiment, the formation of a frequently used artificial flavoring whose
smell resembles the banana or apple namely butyl acetate will be carried out using the techniques
of reflux, solvent extraction and distillation.
III. MATERIALS AND EQUIPMENTS
− 1 round bottom flask − n-Butanol
− 1 Cylinder 100 mL − Acetic acid
− 1 Cylinder 10 mL − Sulfuric acid
− Reflux apparatus with stand, heating − Sodium bicarbonate
mantle and water pump
 − Distillation apparatus with stand,
heating mantle and water pump
− 1 Separatory funnel 250 mL with
stand
− Balance
− 1 Beaker 250 mL
− 3 Beaker 100 mL

III. PROCEDURE
Part A:
1. In a round-bottom flask, weigh and mix 100 mL n-butanol in 75 mL of acetic acid. Carefully
add 5 mL of concentrated sulfuric acid into prepared mixture.
2. Assemble the reflux apparatus, greasing the joints with vacuum grease. Start the water
circulating and bring the mixture to boil under reflux for 60 minutes. Calculate the theoretical
yield of the reaction.
3. Dismantle the reflux. Allow the solution to cool off.

Part B: Extraction and distillation

1. In the hood carefully clamp a 250 mL separatory funnel to a ring stand. Be sure that the
stopcock is closed. Transfer the reaction mixture to the funnel in a careful manner, minding the
excess amount of acid.
2. Continue the liquid-liquid extraction with an appropriate amount of cold water for 3 times. In
details, shake the funnel vigorously and allow the layers to separate in about 10 minutes. Identify
and collect the organic layer containing crude butyl acetate.
3. Prepare a saturated sodium solution by dissolving 16 g of sodium bicarbonate in 200 mL of
water.
4. Wash the crude extract with prepared amount of sodium bicarbonate solution several times in
a separatory funnel. Allow the layers to settle until no visible oil droplets are visible in the
aqueous layer.
5. Collect the washed solution and dispose the remained reaction products while assembling
distillation apparatus. Be careful with carbon dioxide pressure building up in the funnel during
this step.
6. Preweigh a clean and dry Erlenmeyer flask to function as the collection flask.
7. Collect the distillate at butyl ascetic’s boiling point. Weigh the product and calculate the
percentage yield of the ester.

IV. Lab report

Calculate the theoretical yield and the actual yield in the synthesis that has been conducted and
draw conclusions that are related the equilibrium of synthesis reaction. Describe the
characteristics
of the product.

V. Question for discussion

1. Discuss about the importance of obtained ester on a multi-aspects basis and its whole
procedure of synthesis in the lab.
2. Does an industrial scale process of synthesis exist for methyl salicylate? How has it been
constructed and optimized?
3. What are the methods to identify, confirm, and physically or chemically characterize the final
product? Since those are not yet done in the lab session, please provide a few published data
from other articles/reports to illustrate your answer.
 Experiment #9: Solvent Effects in an SN1 Solvolysis Reaction
A Kinetics Study
I. INTRODUCTION
In this experiment, you will not conduct a detailed quantitative kinetics study. Instead,
you will determine the relative rates of the solvolysis of t-butyl chloride in three different
solvent systems (methanol-water, ethanol-water, and acetone-water) and express the results in
graphic form.
In the first two solvent systems, mixtures of organic products are formed because the
alcohols contain hydroxyl (-OH) groups. With the acetone-water mixture, only the water
participates in the solvolysis reaction. Regardless of the solvent system, the inorganic product is
HCl. Note that for each molecule of t-butyl chloride that undergoes reaction, one molecule of
HCl is generated. Because of this 1:1 correspondence, the course of the reaction can be
conveniently followed by measuring the acidity of the reaction mixture.
In this experiment, you will be comparing the time it takes for each of several solvolysis
reactions to reach the same per cent of completion. Because this is a study of relative rates, the
exact percent of completion does not matter as long as all the reactions are carried to the same
point. (Your solvolysis reactions will be carried to only about 5% completion). Detecting the
percent completion point is accomplished by adding a measured amount of NaOH to each
reaction mixture. Under these conditions, each mixture remains alkaline until the NaOH has
been neutralized by the HCl being generated.1 Then, the solvolysis mixture will turn acidic as
additional HCl is generated. We can detect the change from an alkaline solution to one that is
acidic by including phenolphthalein in the solvolysis mixture. When the mixture becomes acidic,
the solution changes from pink to colorless. From a plot of the percent water in each solvent
system versus the time required to reach the phenolphthalein end-point, we can compare the
effects of various solvent systems upon the rate of the SN1 reaction. The experiment as
described is semi-quantitative, and the results cannot be used to calculate the rate constant.
Under the conditions of this experiment, a tertiary alkyl halide can also undergo
elimination by an El or E2 path. In this experiment, these side reactions are ignored. If you carry
out titrations of aliquots of the reaction mixture (instead of adding NaOH directly to the
reaction mixture), side reactions are minimized.
II. MATERIALS AND EQUIPMENTS

− 2 burets − Acetone, 6-10 mL

− Clock − t-butyl chloride, less than 5 mL
− Dropper or disposable pipet − 95% ethanol, 6-10 mL
− 3-5 styrofoam cups − Methanol, 6-10 mL
− 15 test tubes, 13 x 100 mm, with corks − Phenolphthalein indicator
− Thermometer − Sodium bicarbonate
− Tin foil − A few drops 0.5 M sodium hydroxide,
less than 5 mL

III. PROCEDURE:
The solvent systems to be tested are listed in the table below. Because 15 separate
mixtures will be tested, plan to run 3-5 reactions simultaneously. Each reaction requires 5-30
minutes; therefore, your various runs should be planned to overlap.
STOPPING POINTS: Although the experiment could be stopped after any batch has been run, it
is preferable to run all the solvolysis reactions in the same laboratory period. This will ensure
that the same droppers and the same NaOH solution are used.

1. Place 2.0 mL of the appropriate solvent mixture in a clean, labeled test tube. Use the
micropipette to add the proper volume of solvent and the distilled water.
2. Cork the test tubes and place them in a constant-temperature bath for about 5 minutes to
come to thermal equilibrium.
3. Use the waterbath at 30°C for the whole experiment.
To each test tube, add 3 drops of 0.5N sodium hydroxide solution and 1-2 drops of
phenolphthalein indicator.
To one tube at a time, add 3 drops of t-butyl chloride. Mix or shake the contents of the
tube immediately and record the time of the addition to the nearest second. Continue
shaking.
4. When the pink color disappears, again record the time. Repeat this procedure for each of
the
solvent systems.
5. Calculate the elapsed time for reaction in each solvent system to the nearest 0. 1 minute.
6. Plot percent water in each solvent system versus elapsed time. (Place all three plots on
the same graph). Compare the three plots and record your observations and conclusions.