Organic Chemistry Lab Manual

Komar University of Science and
Technology (KUST)
Fall
2015 ORGANIC CHEMISTRY
© The Copy Rights Reserved for Komar University of Science and Technology (KUST),
Sulaymani, Kurdistan-Iraq
Organic Chemistry, Laboratory Manual CHM 2415L
Preface and Acknowledgments
In preparing this laboratory manual, I am thankful to the efforts of KUST

administration, Medical Laboratory Science Department, and staff members who
made this edition possible to be completed with their support.
This laboratory manual shares the outline and pedagogical philosophy of the
textbook, Organic Chemistry, Ed, David Klein. Johns Hopkins University, John
Wiley and Sons, Inc. 2015, as in previous editions, CHM 1410C and 2411C we have
strived for the clearest possible writing in the procedures. The experiments give the
student a meaningful, reliable laboratory experience that consistently works, while
covering the basic principles of general chemistry, organic, and biochemistry.
Three basic goals were followed in all the experiments: (1) the experiments should
illustrate the concepts learned in the classroom; (2) the experiments should be clearly
and concisely written so that students will easily understand the task at hand, will
work with minimal supervision because the manual provides enough information on
experimental procedures, and will be able to perform the experiments in a two-and-a-
half-hour laboratory period; (3) the experiments should not only be simple
demonstrations, but also should contain a sense of discovery.
I anticipate that students will find it inspiring in studying different aspects of

chemistry.
KOMAR UNIVERSITY OF SCIENCE AND TECHNOLOGY (KUST) 1

Contents
1. Common Laboratory Apparatus ……………………………………………………………. 1
2. Chemistry Lab Safety ………………………………………………………………………. 5
2.1 Laboratory Attire ……………………………………………………………………….. 5
2.2 Laboratory Conduct …………………………………………………………………….. 6
2.3 Fire Case ………………………………………………………………………………... 7
3. Writing Lab Reports ………………………………………………………………………... 8
4. Submission of Lab Reports ………………………………………………………………… 8
5. Laboratory Experiments …………………………………………………………………… 9
Experiment 1: SN1: Synthesis of tert-Butyl Chloride …………………………….…………10
Experiment 2: Nucleophilic Substitution Reaction: SN1 versus SN2 …………….………… 15
Experiment 3: Identification of Alcohols and Phenols ……………………………………. 20
Experiment 4: Identification of Aldehydes and Ketones………………………...………… 29
Experiment 5: Synthesis of o-Chlorobenzoic acid …………………..…………………….. 34
Experiment 6: Esterification Reaction: Preparation of Aspirin …………….……………... 39
Experiment 7: Properties of Amines and Amides………………………………………….. 43
Experiment 8: Carbohydrates………………………………………….……….…………... 50
Experiment 9: Hydrolysis of Starch a Polysaccharide……………………………………... 58
Experiment 10: Isolation and Identification of Casein (Protein)…….. ….………………… 62
Experiment 11: Analysis of Lipids. …………………………………………………….….. 70
6. References ……………………………………………………………………………...….. 76
7. Periodic Table of the Elements….…………………………………………………………. 77

1. Common Laboratory Apparatus*
Laboratory apparatus and equipment like electronic scales, glassware, burners, graduated cylinders,
and more, help the scientist conduct experiments, observe data, heat liquids, make measurements, and
more.
Items Name Picture Items Name Picture
Volumetric Flask Test Tubes
Watch Glass Crucible and lid
Dropper Evaporating dish
Funnel Graduated cylinder

Mortar and pestle Beaker
Stirring rod Erlenmeyer Flask
Micropipette Burette
Thermometer Bunsen burner

Wire gauze Clamp
Iron ring Clay triangle
Stand Tong
Wash bottle Safety goggles
Corks Rubber Stopper
Brush Forceps
Test tube holder Spatulas
Suction Filteration Well plate

Centrifuge Oven
Water bath Electronic Lab Scale
Hot plate pH-meter
* Matin J., Martin C. (2012). Catalyst: the Pearson Custom Library for Chemistry: Laboratory
Experiments, Pearson Prentice Hall. ISBN13: 978-0-536-93404-8

2. Chemistry Lab Safety
The chemistry laboratory can be a place of discovery and learning. However, by the very nature of
laboratory work, it can be a place of danger if proper common-sense precautions aren't taken. While
every effort has been made to eliminate the use of explosive, highly toxic, and carcinogenic
substances from the experiments which you will perform, there is a certain unavoidable hazard
associated with the use of a variety of chemicals and glassware. You are expected to learn and adhere
to the following general safety guidelines to ensure a safe laboratory environment for both yourself
and the people you may be working near. Each student will have to pass a lab safety exam before
conducting any lab work to evaluate his/her understanding to the lab rules and safety
precautions. Students who fail in that exam will have to take orientation session about lab
safety before they can start lab work. Additional safety precautions will be announced in class
prior to experiments where a potential danger exists. Students who fail to follow all the safety rules
may be asked to leave the lab or suffer grading penalties.
2.1 Laboratory Attire
1. Safety goggles must be worn at all times while in the laboratory. This rule must be followed
whether you are actually working on an experiment or simply writing in your lab notebook.
You must wear safety goggles provided by the chemistry department.
2. Contact lenses are not allowed. Even when worn under safety goggles, various fumes may
accumulate under the lens and cause serious injuries or blindness.
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.

2.2 Laboratory Conduct

1. Eating, drinking, and smoking are strictly prohibited in the laboratory.
2. No unauthorized experiments are to be performed. If you are curious about trying a

procedure not covered in the experimental procedure, consult with your laboratory
instructor.
3. Coats, backpacks, etc., should not be left on the lab benches and stools. Beware that lab
chemicals can destroy personal possessions.
4. Always wash your hands before leaving lab.
5. Be especially careful of spills around the balance. These electronic devices are extremely
sensitive to corrosion. A brush is kept near the balance so you can brush the balance
thoroughly after each use.
6. Clean any spill near the balance immediately and

report it to instructor.
7. Notify the instructor immediately in case of an

accident.
8. Consider all chemicals to be hazardous as a result

never taste anything, never directly smell the
source of any vapor or gases, by means of your
couple hands, bring a small sample to your nose,
Fig 2.2.1
9. Know what chemicals you are using. Carefully read

the label twice before taking anything from a bottle.
10. Excess reagents are never to be returned to stock bottles. If you take too much, dispose of
the excess.
11. Many common reagents, for example, alcohols and acetone, are highly flammable. Do not
use them anywhere near open flames.
12. Always pour acids into water. If you pour water into acid, the heat of reaction will cause
the water to explode into steam, sometimes violently, and the acid will splatter.

13. If chemicals come into contact with your skin or eyes, flush immediately with copious
amounts of water and consult with your instructor.
14. Never point a test tube or any vessel that you are heating at yourself or your neighbor, Fig.
2.2.2.
15. Dispose of chemicals properly. Waste

containers will be provided and their use will
be explained by your TA. Unless you are
explicitly told otherwise, assume that only
water may be put in the lab sinks.
16. Clean up all broken glassware immediately

and dispose of the broken glass properly.
17. Never leave burners unattended. Turn them off whenever you leave your workstation. Be
sure that the gas is shut off at the bench rack when you leave the lab.
18. Beware of hot glass--it looks exactly like cold glass.
2.3 Fire Case
1. In the event of fire, do not panic.
2. If a small portion of your clothes catches fire, the fire may be extinguished by patting it out.
3. Never use a fire extinguisher on a person. Carbon dioxide fire extinguisher is extremely
cold and may cause shock to the person, frostbites, or harm to the eyes.
4. If a fire should occur in a breaker or some other container, cover it with a glass dish or other
flame-retardant item.

3. Writing Lab Reports
The lab report for this course will be represented by a designed report sheet prepared by the course
instructor and accompanying with each experiment. This sheet will illustrate the understanding of
the experiment via answering some questions related to the work and it also summarize the data via
tables with simple calculations whenever it is needed.
4. Submission of Lab Reports
The lab report sheet of previous week experiment should be submitted to the lab assistant or the
course instructor at the beginning of the next experiment lab work. Late submission will cause
deduction 1 grade of your total report sheet grade for each one hour lateness after the submission
due date that announced by the instructor. No report sheets will be collected after 10 hours from
the submission date. Also individual submission is not accepted, each group should submit one
report sheet for their work.

5. Laboratory Experiments

Experiment 1
SN1 Synthesis of tert-Butyl Chloride
Objectives
1. Synthesize tert-butyl chloride via an SN1 reaction.

2. Confirm the presence of a tertiary alkyl halide using the silver nitrate test.
Introduction
3. Objectives
Alkyl halides can be prepared from their corresponding alcohols via an acid catalyzed substitution
reaction. The mechanism of these acid catalyzed substitution reactions are labeled as SN1
(substitution, nucleophilic, unimolecular) and SN2 (substitution, nucleophilic, bimolecular). Tertiary
alcohols follow the SN1 route, primary alcohols follow the SN2 route, and secondary alcohols can
follow either path. Under acidic conditions, the mechanism (Figure 1.1) of the SN1 reaction involves
rapid protonation of the alcohol, followed by the loss of water as the rate-determining step. This
generates a relatively stable carbocation which is then attacked by the halide ion to form the alkyl
halide. SN1 reactions are favored by tertiary alcohols because they tend to form more stable
carbocations.
Figure 1.1: Conversion of t-butanol into t-butyl chloride by an SN1 mechanism
Confirmation of your product, tert-butyl chloride, can be performed by reacting the product with a
solution of silver nitrate in ethanol. The tertiary alkyl halide will react by means of an SN1 mechanism
with the silver nitrate to form an insoluble silver halide. The appearance of a precipitate indicates a
positive result indicating the presence of a tertiary alkyl halide.

Materials
Table 1.1: Chemicals and supplies
Chemicals Supplies
concentrated HCl 25 mL graduated cylinder
tert-Butanol, reagent grade 10 mL glass vials and caps (2)
Saturated NaHCO3 solution 10 mL graduated cylinder
Saturated NaCl solution 50 mL beaker
Na2SO4, anhydrous 250 mL beaker
DI water 125 mL Erlenmeyer flask
Ice Separatory funnel apparatus
1% ethanolic-AgNO3 solution Glass funnel
tert-Butyl chloride (product from Part I) Disposable droppers
3 test tubes
Test tube rack
Procedure
Part I: Synthesis of tert-butyl chloride
Note: Conduct step 2-3 in a fume hood and leave it inside the hood until you finish setting
up the Separatory funnel apparatus in step 4.
1. Prepare an ice-water bath in a 250 mL beaker.
2. Measure 15 mL of concentrated HCl using a graduated cylinder and carefully transfer to a 50 mL

Erlenmeyer flask.
CAUTION: Handle the HCl with care. It can cause painful burns if it comes in contact with
the skin.
3. Cool the Erlenmeyer flask containing the acid in the ice-water bath that you prepared in step 1.
4. Set up the reaction in a Separatory funnel. Figure 1.2:
a. Clamp the support ring onto a ring stand, place the Separatory
funnel into the ring, and insert a funnel.
b. Carefully transfer the cooled acid to the Separatory funnel.
c. Weigh 4.0 g of tert-butanol using a beaker and transfer to the
Separatory funnel. Record the mass in the report sheet.
Figure 1.2: Separatory Funnel Set up

5. Swirl the solution in the Separatory funnel for five minutes.
Note: Leave the funnel uncapped as you swirl to allow the gases to escape.
6. Cap the Separatory funnel, invert it, and immediately open the stopcock to release the pressure.
Figure 1.3.
Note: When venting the funnel, point the

tip in a direction where there’s no one
and open the stopcock to release the
pressure.
7. Shake the funnel for 15 minutes. Vent

occasionally to prevent pressure buildup.
8. Place the funnel in the support ring and

allow the solvent and aqueous layers to
separate. Leave the funnel uncapped.
9. Drain the lower aqueous layer into a 125

mL Erlenmeyer flask. Be sure to label the
flask as waste.
Figure 1.3: Venting the Separatory
10. Prepare to wash the solution: Funnel
a. Wash the organic layer with 30 mL of
a saturated sodium bicarbonate solution. Add the sodium bicarbonate solution slowly. Swirl
the funnel several times until the bubbling has stopped.
b. Cap the funnel, invert, shake, and immediately vent.
c. Place the uncapped funnel in the support ring and allow the layers to separate.
d. Drain the aqueous layer into the waste beaker.
11. Repeat Step 10 with using another 30 mL of a saturated sodium chloride solution.
12. Prepare to dry the product:

a. Place a pea-sized amount of anhydrous sodium sulfate in a 10 mL vial.
b. Transfer the organic layer into the vial and swirl until clumps form.
c. Cap the vial and wait five minutes to allow the sodium sulfate to dry the product.
13. While waiting, weigh a clean and dry 10 mL vial. Record the mass in your report sheet.
14. Carefully pipet the product into the clean vial and weigh. Be careful not to transfer any of the
sodium sulfate. Record the mass in your report sheet.

Part II Silver Nitrate Test for Tertiary Alkyl Halides
1. Set up the reaction.

a. Obtain two test tubes.
b. Pipet 10 drops of distilled water into test tube 1.
c. Pipet 10 drops of your product into test tube 2.
2. Add approximately 1 mL of the 1% ethanolic silver nitrate solution to each test tube. Swirl each
test tube to mix the contents. The appearance of a white precipitate indicates the presence of a 3°
halide. Record your observations in the report sheet.
Note: Avoid getting the AgNO3 solution on your skin.

Report Sheet
Grade Group Names:
Date:
1. Collecting Data:
Table 1.2: Part I: Synthesis of tert-Butyl Chloride
Mass of tert-Butanol g
Mass of vial g
Mass of vial and product g
Mass of product g
Theoretical mass of tert-Butyl Chloride g
Percent yield %
Table 1.3: Part II: Silver nitrate test for alkyl halides
Test Compound Observation
1 H2O
2 Product
2. Answer the following questions:

a. Write an SN1 mechanism of the reaction?
b. Why it is necessary to conduct the reaction under acidic conditions?
c. Write the molecular chemical equation for the reaction between alcohol and HCl.

Experiment 2
Nucleophilic Substitution Reaction: SN1 versus SN2
Objectives
1. Learn how variation in organo-halide structure affects the rate of SN1 and SN2 reactions
2. Observe the reaction rate by measuring the time required for a visible change to occur (formation
of a precipitate).
Introduction
Nucleophilic substitution is one of the most useful and well-studied class of organic reactions.
These reactions can occur by a range of mechanisms. SN2 and SN1 are the extremes.
The SN2 reaction occurs in a single step. The nucleophile enters as the leaving group —
usually a halide ion — departs. The reaction displays second-order kinetics; its rate is
proportional to the concentration of the organo-halide and the nucleophile.
In the SN1 reaction loss of the leaving group occurs first to generate a carbocation
intermediate. The carbocation then captures a nucleophile, often the solvent (followed by
proton transfer to produce the final neutral product). In this case the reaction is called a
solvolysis.
Because the first step is rate-determining, the SN1 reaction displays first-order kinetics; its
rate depends only on the concentration of the organo-halide. It will be easier to remember
which label goes with which mechanism if you associate the "1" in SN1 with carbocation
rather than with the kinetic order of the reaction. (Perhaps this should be called SNCarbocat or
SNC+.)
Which mechanism occurs under a certain set of conditions and how fast it occurs depend on a
variety of factors. The structure of the organo-halide, the leaving group, the nucleophile, and
the solvent can all play a role.
An assortment of alkyl, alkenyl, and aromatic chlorides and bromides will be available.
To encourage an SN2 reaction mechanism you will use a solution of NaI in acetone. Iodide is
a good nucleophile, and if it displaces bromide or chloride, NaBr or NaCl will precipitate
(these are much less soluble in acetone than NaI). To encourage an SN1 reaction mechanism
you will use a solution of AgNO3 in ethanol. Ethanol is a polar protic solvent and can
promote ionization of certain organo-halides. If halide ion is released a precipitate of AgCl or
AgBr will form.

Materials

Chemicals Supplies
1 M NaI/acetone Test tube (7)
1-bromobutane 10 mL Graduated cylinder (2)
bromobenzene Disposable droppers
2-bromo-2-methylpropane Warm water bath 40-50°C
Allyl bromide
Allyl chloride
Bromo benzene
Benzyl chloride
2% AgNO3/EtOH
Note: The TA should prepare necessary solutions before lab starts.
Procedure
Part I: SN2 Reaction

1. Bring 3 small, dry, clean test tubes and add to each test tube 4 drops of 1-bromobutane,
bromobenzene, 2-bromo-2-methylpropane respectively.
2. Add 1 mL of NaI/acetone to each test tube and record the time for precipitation to form in table
2.1 in the report sheet.
3. If no precipitate form after about 3 minutes, warm the test tubes to 40-50°C and record the time
required for precipitation in table 2.1. You will call a test tube (unreactive) if no reaction visible
after about 10 minutes.
4. Complete table 2.1 based on what you leaned in class.
5. Bring 4 small, dry, clean test tubes and add to each test tube 4 drops of the allyl and bezyl
halides that’s available in the lab and repeat steps 2-5. see the note
Note: the allyl and benzyl halides should be tested in the hood because they are strong
lachrymators.
Part II: SN1 Reaction

Repeat step 1-5 using AgNO3/EtOH solution and record your observation in table 2.3

Report Sheet
Grade Group Names:
Date:
Table 2.2: Recording observation for SN2 reaction

Adding 1 M *Predict the relative Observed time for
Structure of organo-halide
NaI/acetone reactivity forming ppt
1-
bromobutan
e
2-
bromobutan
e
2-bromo-2-
methylprop
ane
Allyl
bromide
Allyl
chloride
Bromo
benzene
Benzyl
chloride
*use the words: very slow, slow, fast, very fast…etc

Report Sheet 2
Table 2.3: Recording observation for SN1 reaction

organo- Structure of Adding 2% *Predict the relative Observed time for
halide organo-halide AgNO3/EtOH reactivity forming ppt
1-
bromobutane
bromobenze
ne
2-bromo-2-
methylpropa
ne
Allyl
bromide
Allyl
chloride
Bromo
benzene
Benzyl
chloride
*use the words: very slow, slow, fast, very fast...etc

Report Sheet 3
Answer the following questions:

1. What was the effect of substitution at the C undergoing nucleophilic attack, in particular, 1° vs 2°
vs 3° alkyl?
2. Do the allylic and bezylic halides fit this pattern? If not, suggest an explanation.
3. What was the effect (if any) of changing the leaving group?

Experiment 3
Identification of Alcohols and Phenols
Objectives
1. To learn characteristic chemical reactions of alcohols and phenols.

2. To use these chemical characteristics for identification of an organic compound.
Introduction
Specific groups of atoms in an organic molecule can determine its physical and chemical properties.
These groups are referred to as functional groups. Organic compounds which contain the functional
group _OH, the hydroxyl group, are called alcohols.
Alcohols are important commercially and include uses as solvents, drugs, and disinfectants. The most
widely used alcohols are methanol or methyl alcohol, CH3OH, ethanol or ethyl alcohol, CH3CH2OH,
and 2-propanol or isopropyl alcohol, (CH3)2CHOH.
Methyl alcohol is found in automotive products such as antifreeze and “dry gas.” Ethyl alcohol is
used as a solvent for drugs and chemicals, but is more popularly known for its effects as an alcoholic
beverage. Isopropyl alcohol, also known as “rubbing alcohol,” is an antiseptic.
Alcohols may be classified as either primary, secondary, or tertiary:
Note that the classification depends on the number of carbon-containing groups, R (alkyl or
aromatic), attached to the carbon bearing the hydroxyl group. Examples of each type are as follows:

Phenols bear a close resemblance to alcohols structurally since the hydroxyl group is present.
However, since the _OH group is bonded directly to a carbon that is part of an aromatic ring, the
chemistry is quite different from that of alcohols. Phenols are more acidic than alcohols; concentrated
solutions of the compound phenol are quite toxic and can cause severe skin burns. Phenol derivatives
are found in medicines; for example, thymol is used to kill fungi and hookworms.
Physical Properties
Since the hydroxyl group is present in alcohols and phenols, these compounds are polar. The polarity
of the hydroxyl group, coupled with its ability to form hydrogen bonds, enables many alcohols and
phenols to mix with water. Since these compounds also contain nonpolar portions, they show
additional solubility in many organic solvents, such as dichloromethane and diethyl ether.
Chemical Properties
The chemical behavior of the different classes of alcohols and of phenols can be used as a means of
identification. Quick, simple tests that can be carried out in test tubes will be performed.
1. Lucas test. This test is used to distinguish between water-soluble primary, secondary, and tertiary
alcohols. Lucas reagent is a mixture of zinc chloride, ZnCl2, in concentrated HCl. Upon addition
of this reagent, a tertiary alcohol reacts rapidly and immediately gives an insoluble white layer. A
secondary alcohol reacts slowly and, after heating slightly, gives the white layer within 10 min. A
primary alcohol does not react.

Any formation of a heterogeneous phase or appearance of an emulsion is a positive test.
2. Chromic acid test. This test is able to distinguish primary and secondary alcohols from tertiary
alcohols. Using acidified dichromate solution, primary alcohols are oxidized to carboxylic acids;
secondary alcohols are oxidized to ketones; tertiary alcohols are not oxidized. (Note that in those
alcohols which are oxidized, the carbon that has the hydroxyl group loses a hydrogen.) In the
oxidation, the brown-red color of the chromic acid changes to a blue-green solution. Phenols are
oxidized to nondescript brown tarry masses.
3. Iodoform test. This test is more specific than the previous two tests. Only ethanol (ethyl alcohol)
and alcohols with the part structure CH3CH(OH) react. These alcohols react with iodine in
aqueous sodium hydroxide to give the yellow precipitate iodoform.
Ph
enol
s
also
react
unde
r these conditions. With phenol, the yellow precipitate triiodophenol forms.

4. Acidity of phenol. Phenol is also called carbolic acid. Phenol is an acid and will react with base;
thus phenols readily dissolve in base solutions. In contrast, alcohols are not acidic.
5. Iron(III)chloride test. Addition of aqueous iron(III) chloride to a phenol gives a colored solution.
Depending on the structure of the phenol, the color can vary from green to purple.

Materials

Chemicals Supplies
1-butanol 10 test tubes
2-butanol pH paper
2-methyl-2-propanol Big beaker (serve as water bath 60°C)
Phenol Hot plate
Dioxane Glass rod
6M NaOH
I2/KI solution
Lucas reagent
Acetone
1 M Chromic acid, H2CrO4
1M Iron (III) chloride, FeCl3
Unknown
Note: The TA should prepare the necessary solutions before lab starts
Procedure
Part I: Physical Properties of Alcohols and phenols

1. Bring 4 separated test tubes and label them: 1-butanol, 2-butanol, 2-methyl-2-propanol, and
unknown respectively then add 10 drops of each sample.
2. Add 3 mL of distilled water to each test tube.
3. Into a separate test tube, place 2 mL of a prepared water solution of phenol. Record your
observations as soluble or insoluble sample in table 3.1 of the Report Sheet.
4. Test the pH of each of the aqueous solutions. Do the test by first dipping a clean glass rod into the
solutions and then transferring a drop of liquid to pH paper and read the value of the pH by
comparing the color to the chart on the dispenser. Record the results in table 3.1.

Part II: Chemical Properties of Alcohols and phenols
A. Iodoform Test
1. Label 5 clean, dry test tubes as 1-butanol, 2-butanol, 2-methyl-2-propanol, phenol, and unknown
respectively then add 5 drops of each sample to be tested.
2. Add to each test tube 2 mL of water. If the compound is not soluble, add dioxane (dropwise) until
the solution is homogeneous.
3. Add to each test tube (dropwise) 2 mL of 6 M NaOH; tap the test tube with your finger to mix.
4. Warmed the mixture in each test tube in a 60°C water bath for 5 minutes, and then add dropwise
(with shaking) I2/KI solution until the solution becomes brown (approx. 25 drops).
Note: while it is in the water bath and if the color is fade, add more drops of I2/KI solution until
the dark color persists for 2 min.
5. Add 6 M NaOH (dropwise) until the solution becomes colorless. Keep the test tubes in the warm
water bath for 5 min.
6. Remove the test tubes from the water bath, let it cool, and look for a light yellow precipitate.
Record your observations in table 3.1.
Note: If the formation of the yellow precipitate tends to be slow. Put these test tubes a side and
make your observations at the end of the lab.
B. Lucas test
A. Place 5 drops of each sample into a clean, dry test tubes and labeled them as you did before.
B. Add 1 mL of Lucas reagent; mix well by stoppering each test tube with a cork, tapping the test
tube sharply with your finger for a few seconds to mix; remove the cork after mixing and allow
each test tube to stand for 6 min. Look carefully for any cloudiness that may develop during this
time period.
Note: If there is no cloudiness after 10 min., warm the test tubes that are clear for 15 min. in a
60°C water bath. Record your observations in table 3.1.

C. Chromic acid test
CAUTION! Chromic acid is toxic and corrosive. Handle with care and promptly wash any spill.
Use gloves with this reagent
1. Repeat step 1 in part II (B).
2. To each test tube add 10 drops of acetone and 2 drops of 1M chromic acid. Place the test tubes in
a 60°C water bath for 5 min. observe the color of each solution and Record your observations in
table 3.1.
Note: you will get a positive test when the brown-red will be lost and the blue-green color is
formed.
D. Iron(III) chloride test
1. Repeat step 1 in part II (B).

2. Add 2 drops of 1M iron(III) chloride solution to each test tube and observe any color changes in
each solution. Record your observations in table 3.1.
Note: a purple color indicates the presence of a phenol

Report Sheet 1
Grade Group Names:
Date:
Table 3.2: Recording Observation

2-methy-2-
Test 1-butanol 2-butanol Phenol Unknown
propanol
Solubility in
H2O
pH test
Iodoform
Lucas
H2CrO4 acid
FeCl3
Identity of unknown:
Unknown no. ______________. The unknown compound is ______________.

Report Sheet 2

1. Write the structure of the major organic product expected from each of the following reactions. If
no reaction is expected write “No Reaction.”
2. A student had two unknown liquid alcohols. Unknown A gave a blue-green color with chromic
acid and formed a precipitate after heating for 10 min. with Lucas reagent. Unknown B showed
no color change with chromic acid but formed an immediate precipitate with Lucas reagent. To
which alcohol classes do alcohols A and B belong?
3. What simple test can be used to distinguish between an alcohol and a phenol?

Experiment 4
Identification of Aldehydes and Ketones
Objectives
1. To learn the chemical and physical characteristics of aldehydes and ketones.

2. To use these chemical characteristics in simple tests to distinguish between examples of
aldehydes and ketones.
Introduction
Aldehydes and ketones are representative of compounds which possess the carbonyl group:
The carbonyl group
Aldehydes have at least one hydrogen attached to the carbonyl carbon; in ketones, no hydrogen is
directly attached to the carbonyl carbon, only carbon containing R-groups:
Aldehydes and ketones of low molecular weight have commercial importance. Many others occur
naturally.

Classification Tests
1. Chromic acid test. Aldehydes are oxidized to carboxylic acids by chromic acid; ketones are
not oxidized. A positive test results in the formation of a blue-green solution from the brown-
red color of chromic acid.
2. Benedicts’ test. Is a very sensitive test in which the oxidation of aldehyde is accompanied by
simultaneous reduction of copper(II) ion Cu2+ to a highly colored precipitate of copper(I) oxide,
Cu2O. The appearance of the color product signals the oxidation of an aldehyde to the carboxylic
acid. Most often the copper(I) oxide, Cu2O forms as a dark red precipitate but it can also produce
a yellow or orange-yellow color too. Any red-orange-yellow color indicate the presence of an
aldehyde.
The Benedict reagent is basic (NaOH) solution of copper sulfate, CuSO4. Because the test
solution is basic, the sodium salt of the carboxylic acid is produced:
3. Iodoform test. Methyl ketones give the yellow precipitate iodoform when reacted with iodine in
aqueous sodium hydroxide.

Materials
4.1: Chemicals and supplies

Chemicals Supplies
Propanal 6 test tubes with lid or cork or Parafilm
2-methyl propanal 400 mL beaker serve as boiling water bath
n-butyl aldehyde rack
Glucose 10 mL graduated cylinder
Acetone
cyclohexanone
Ethanol
2-propanol
4-hyptanone
*1 M Chromic acid
**Benedict reagent
**Benedict reagent: Solution 1: dissolve 1.73 g hydrated CuSO4 crystals in 20 mL distilled water. Solution 2:
dissolve 10 g anhydrous Na2CO3 in 70 mL distilled water. Add solution 1 to solution 2 and complete the volume
to 100 ml volumetric flask.
*TA should prepare the necessary solutions
Procedure
Part I: Physical properties of Aldehydes and Ketones
A. Solubility of Aldehydes and Ketones
1. Label 6 test tubes as propanal, 2-methyl propanal, n-butyl aldehyde, glucose, acetone, and 4-
hyptanone respectively then add 10 drops of each sample to be tested.
Note: glucose is solid, you should add small amount of glucose particles in the test tube
equal to the size of the head of a match.
2. Add 1 mL of distilled water to each test tube and seal each test tube with a cork or Parafilm
and shake each mixture thoroughly, then set it aside in a test tube rack.
3. Wait for 5 minutes and record your observation in table 4.2 in the report sheet
Note: if the sample is insoluble in water you should observe either a cloudy mixture or two
separated layers.

Part II: Chemical Properties of Aldehydes and Ketones
A. Chromic Acid Test

1. Label 5 clean test tubes as propanal, 2-propanol, ethanol, acetone (Control), and 4-hyptanone
respectively then add 4 drops of each sample to be tested.
2. Add 1 mL of acetone to each test tube (except the acetone test tube
CAUTION! Chromic acid is toxic and corrosive. Handle with care and promptly wash any
spill. Use gloves with this reagent
3. Add one drop of chromic acid to the acetone test tube only (the color of the mixture should
remain orange).
Note: if the color change, you should repeat step 3 in another clean test tube
4. Seal each test tube with a cork or Parafilm and shake each mixture thoroughly.
5. Add one drop of chromic acid to each test tube and shake the mixture again and observe any
change in color within 10 seconds and record your Observation in table 4.3 in the report sheet.
6. Set the test tubes aside in a test tube rack for 5 minutes for more evident observation.
B. Benedict Test
1. Put 200 mL water in a 400 mL beaker and heat it till boiling.
2. Label 5 clean test tubes as distilled water (Control), propanal, ethanol, glucose, and acetone
respectively
3. Add 2 mL of Benedict reagent to each test tube.
4. Add 1 mL of the substance to each of the labeled samples to be tested.
Note: glucose is solid, you should add small amount of glucose particles in the test tube
equal to the size of the head of a match.
5. Seal each test tube with a clean cork or Parafilm and mix thoroughly.
6. Place each labeled in a boiling water bath for 10 minutes.
7. Remove and place the test tubes in a test tube rack, as the test tubes cool check for a dark red,
orange, brown, yellow precipitate and record the observation in table 4.3 in the report sheet

Report Sheet 1 Grade Group Names:

Date:
Table 4.2: Test the Solubility of the substances in water

2-heptanone Propanal 2-methylpropanal Heptanal Glucose Acetone
Table 4.3: Chemical Properties of Aldehydes and Ketones

Test Acetone Ethanol 2-propanol Propanal 2-heptanone Glucose
Chromic acid ---
Benedict Reagent --- ---
Answer the following question:

1. What happens to an aldehyde in its reaction with chromic acid?
2. What is the result of oxidation of 2-propanol?
3. What kind of results do you see when the following compounds are mixed together with the given
test solution?
Benedict reagent

Experiment 5
Synthesis of O-Chlorobenzoic acid
Objectives
1. Carry out a diazotization reaction on anthranilic acid to prepare the corresponding diazonium salt.
2. Carry out a Sandmeyer reaction on the diazonium salt to prepare an aryl halide.
Introduction
Primary arylamines react with nitrous acid, HNO2, to yield stable arenediazonium salts,
Ar-N+≡N--X,a process called diazotization reaction. Alkylamines also react with nitrous acid, but the
corresponding alkanediazonium products are so reactive they can’t be isolated. Instead they lose
nitrogen instantly to yield carbocations. The analogous loss of N2 from an arenediazonium ion to
yield an aryl cation is disfavored by the instability of the cation.
Arene diazonioum salts are useful because the diazonio group (N2) can be replaced by a nucleophile
in a substitution reaction. Many different nucleophiles – halide, hydride, cyanide, and hydroxide
among others – react with arenediazonium salts, yielding many different kinds of substituted
benzenes. The overall sequence of (1) nitration, (2) reduction, (3) diazotization, (4) nucleophilic
substitution is perhaps the single most versatile method of aromatic substitution.
Aryl chlorides and bromides are prepared by reaction of an arenediazonium salt with the
corresponding copper(I) halide, CuX, a process called the Sandmeyer reaction.
Mechanism
Mechanistically, the diazonio replacement reaction occur through radical rather than polar pathways.
In the presence of a copper(I) compound, for instance, it’s thought that the arenediazonium ion is first
converted to an aryl radical plus copper(II), followed by subsequent reaction to give product plus
regenerated copper(I) catalyst.

Reaction

Materials
Table 5.1: Chemicals and Supplies

Chemical Supplies
CuCl2.H2O 125 mL Erlenmeyer Flask
Copper metal (small pieces) 250 mL Erlenmeyer Flask
N-Phenyl Anthranilic acid Salt-ice water bath
Conc HCl Hot plate
Cold conc HNO2 600 mL beaker
NaNO2 Glass rod and thermometer
Büchner funnel, filter flask, filter paper (Fig 5.1)
50 mL graduated cylinder
Procedure
Part I: Preparing cold solution of CuCl2
1. In a 125 mL Erlenmeyer flask weigh 4.7 g CuCl2.2H2O.
2. Add 20 mL of water to step 1 and stir until complete dissolution.
3. Add 15 mL of concentrated HCl to step 1 and 3.5 g copper metal and heat the mixture until it
boils gently.
4. Keep boiling for about 15 min time after which a decoloration should be observed. Meanwhile,
carry out the following process:
Part II: Diazotization process

1. Measure 10 mL concentrated HCl and transfer it to a 250 mL Erlenmeyer flask and add 50 mL
water.
2. Add 5.5 g N-Phenyl Anthranilic acid acid to step 1 and Dissolve it by heating slightly and then
cool the solution in an ice-salt bath and leave it there.
3. While monitoring the temperature, in a beaker prepare a solution of NaNO2 by dissolving 2.8 g of
NaNO2 in 10 mL water
4. Add dropwise of solution in step 3 to the anthranilic acid solution in the 250 mL Erlenmeyer
flask.
Note: The temperature of the mixture should not exceed 0.0°C.
5. Once the addition is over, keep the Erlenmeyer flask in the ice-salt bath.

Part III: Sandmeyer reaction

1. In a 600 mL beaker, add the CuCl2 solution from part I and cool it down quickly below 0 °C.
2. Add the diazonium salt from part II gradually while stirring vigorously with a glass rod.
Note: A large amount of foam is produced due to the release of nitrogen gas.
3. Keep stirring for 30 min. Then, filter over a Büchner funnel, figure 5.1, and wash the precipitate,
first with cold ~8 M HNO2, then with cold water until the filtrate gets colorless.
4. Dry the precipitate over vacuum.
5. Weigh the mass of crude product (o-chlorobenzoic acid) obtained.
Figure 5.1: Apparatus for suction filtration

Report Sheet 1
Grade Group Names:
Date:
Mass of pure o-chlorobenzoic acid expected: 6.28 g
Mass of crude o-chlorobenzoic acid obtained: _______________ g
𝑀𝑎𝑠𝑠 𝑜𝑓 𝑐𝑟𝑢𝑑𝑒 𝑝𝑟𝑜𝑑𝑢𝑐𝑡 𝑜𝑏𝑡𝑎𝑖𝑛𝑒𝑑

Percent yield in crude product = × 100 =
𝑀𝑎𝑠𝑠 𝑜𝑓 𝑝𝑢𝑟𝑒 𝑝𝑟𝑜𝑑𝑢𝑐𝑡 𝑒𝑥𝑝𝑒𝑐𝑡𝑒𝑑
= _______________ × 100
= %
1. What is the purpose of washing the anthranilic acid with HNO2 solution?
2. Give another example of nucleophile that could react with the diazonium salt. Write the equation.

Experiment 6
Esterification Reaction: Preparation of Aspirin
Objectives
1. Synthesize aspirin from its corresponding acid anhydride and alcohol
2. Compare two different synthetic routes for the preparation of esters
Introduction
The classic synthesis of esters is the Fischer-Speier esterification, employed in experiment 1.

However, several other methods are available, one being often favored other another depending on
the problems needing to be tackled. The method used in this experiment is the alcoholysis of an acid
anhydride.
Alcohols react with acyl chlorides or acid anhydrides to give esters:
These reactions are irreversible, thus simplifying workup. Since acyl chlorides and acid anhydrides
react also with water, anhydrous conditions are preferred. The analogous acylation of amines that
produces amides is less sensitive towards water because amines are stronger nucleophiles and react
more rapidly.

General Mechanism
Reaction

Materials
Chemicals Supplies
salicylic acid (2 g) 50 mL beaker
acetic anhydride (3 mL) thermometer
conc. sulfuric acid (1 drop) glass rod
Büchner funnel, filter paper
melting point apparatus
Water bath (bain-marie) 50-60°C
10 mL graduated cylinder
Procedure
1. Weight 2.0 g of salicylic acid using a 50 mL beaker.
2. Measure 3 mL of acetic anhydride using the graduated cylinder and add it to step 1.
3. Add 1 drop of concentrated sulfuric acid to the mixture in the beaker with stirring.
4. Heat the mixture using the water bath 50-60°C for 15 minutes with continuous stirring using a
glass rod.
5. Add 35 mL of distilled water, swirl the mixture and filter it using vacuum filtration (refer to
figure 5.1)
6. Weigh the mass of crude product (aspirin) obtained

Report Sheet 1
Grade Group Names:
Date:
Mass of pure aspirin expected: 2.61 g
Mass of crude aspirin obtained: _______________ g
𝑀𝑎𝑠𝑠 𝑜𝑓 𝑐𝑟𝑢𝑑𝑒 𝑝𝑟𝑜𝑑𝑢𝑐𝑡 𝑜𝑏𝑡𝑎𝑖𝑛𝑒𝑑

Percent yield in crude product = × 100 =
𝑀𝑎𝑠𝑠 𝑜𝑓 𝑝𝑢𝑟𝑒 𝑝𝑟𝑜𝑑𝑢𝑐𝑡 𝑒𝑥𝑝𝑒𝑐𝑡𝑒𝑑
= _______________ × 100
= %
Give an alternative method of synthesis of aspirin, using salicylic acid as a stating material. Give the
mechanism.

Experiment 7
Properties of Amine and Amides
Objectives
1. To show some physical and chemical properties of amines and amides.
2. To demonstrate the hydrolysis of amides.
Introduction
Amines and amides are two classes of organic compounds which contain nitrogen. Amines behave as
organic bases and may be considered as derivatives of ammonia. Amides are compounds which have
a carbonyl group connected to a nitrogen atom and are neutral. In this experiment, you will learn
about the physical and chemical properties of some members of the amine and amide families.
If the hydrogens of ammonia are replaced by alkyl or aryl groups, amines result.
Depending on the number of carbon atoms bonded directly to nitrogen, amines are classified as either
primary (one carbon atom), secondary (two carbon atoms), or tertiary (three carbon atoms).
There are a number of similarities between ammonia and amines that carry beyond the structure.
Consider odor. The smell of amines resembles that of ammonia but is not as sharp. However, amines
can be quite pungent. Anyone handling or working with raw fish knows how strong the amine odor
can be, since raw fish contains low-molecular-weight amines such as dimethylamine and
trimethylamine. Other amines associated with decaying flesh have names suggestive of their odors:
putrescine and cadaverine.
The solubility of low-molecular-weight amines in water is high. In general, if the total number of
carbons attached to nitrogen is four or less, the amine is water soluble; amines with a carbon content
greater than four are water insoluble. However, all amines are soluble in organic solvents such as
diethyl ether or dichloromethane.

Since amines are organic bases, water solutions show weakly basic properties. If the basicity of
aliphatic amines and aromatic amines are compared to ammonia, aliphatic amines are stronger than
ammonia, while aromatic amines are weaker. Amines characteristically react with acids to form
ammonium salts; the non-bonded electron pair on nitrogen bonds the hydrogen ion.
If an amine is insoluble, reaction with an acid produces a water-soluble salt. Since ammonium salts
are water soluble, many drugs containing amines are prepared as ammonium salts. After working
with fish in the kitchen, a convenient way to rid one’s hands of fish odor is to rub a freshly cut lemon
over the hands. The citric acid found in the lemon reacts with the amines found on the fish; a salt
forms which can be easily rinsed away with water.
Amides are carboxylic acid derivatives. The amide group is recognized by the nitrogen connected to
the carbonyl group. Amides are neutral compounds; the electrons are delocalized into the carbonyl
(resonance) and thus, are not available to bond to a hydrogen ion.
Under suitable conditions, amide formation can take place between an amine and a carboxylic acid,
an acyl halide, or an acid anhydride. Along with ammonia, primary and secondary amines yield
amides with carboxylic acids or derivatives.
Hydrolysis of amides can take place in either acid or base. Primary amides hydrolyze in acid to
ammonium salts and carboxylic acids. Neutralization of the acid and ammonium salts releases
ammonia which can be detected by odor or by litmus.

Secondary and tertiary amides would release the corresponding alkyl ammonium salts which, when
neutralized, would yield the amine.
In base, primary amides hydrolyze to carboxylic acid salts and ammonia. The presence of ammonia
(or amine from corresponding amides) can be detected similarly by odor or litmus. The carboxylic
acid would be generated by neutralization with acid.
Materials

Chemicals supplies
6 M ammonia 5 small test tubes
Triethylamine Watch glass
Aniline 1 large test tubes
N,N-dimethylaniline Boiling water bath
N,N-Dimethyl Acetamide pH-paper
6 M HCl Ice-water bath
Diethyl ether 10 mL graduated cylinder
Conc. HCl
6 M H2SO4
6 M NaOH
Note: TA must prepare the necessary solutions before lab starts.

Procedure
CAUTION!
Amines are toxic chemicals. Avoid excessive inhaling of the vapors and use gloves to avoid direct
skin contact. Anilines are more toxic than aliphatic amines and are readily absorbed through the
skin. Wash any amine or aniline spill with large quantities of water. Diethyl ether (ether) is
extremely flammable. Be certain there are NO open flames in the immediate area.
Part I: Properties of Amines
1. Label 5 clean, dry test tubes as NH3, Triethylamine, Aniline, N,N-Dimethylaniline, and N,N-
Dimethyl Acetamide respectively then add 5 drops (or about 0.1 g if solid) of each sample to
be tested.
2. Add 2 mL of distilled water to each of the labeled test tubes. Mix thoroughly by sharply
tapping the test tube with your finger. Record your observation in table 7.2 in the report sheet
whether the amines are soluble or insoluble.
3. Take a glass rod and carefully dip one end of the glass rod into a solution and touch a piece of
pH paper. Between each test, be sure to clean and dry the glass rod. Record the pH of each
solution.
4. Carefully add 2 mL of 6 M HCl to each test tube. Mix thoroughly by sharply tapping the test
tube with your finger. Record your observation in table 7.2 in the report Sheet whether the
amines are soluble or insoluble.
5. Repeat step 1 and Add 2 mL of diethyl ether (ether) to each test tube. Stopper with a cork and
mix thoroughly by shaking. Record the observed solubility in table 7.2.
6. Carefully place on a watch glass, side-by-side, without touching, a drop of triethylamine and a
drop of concentrated HCl. Record your observations in the report sheet.

Part II: Hydrolysis of acetamide

1. In a large clean, dry test tubes put about 0.5 g of N,N-Dimethyl Acetamide.
2. Measure 5 mL of 6 M H2SO4 using a graduated cylinder and add it to step 1 to dissolve the
acetamide.
3. Heat the solution in a boiling water bath for 5 min.
4. Hold a small strip of moist pH paper over the mouth of the test tube; observe any changes in
color; record the pH reading in table 7.3.
5. Remove the test tube from the water bath, holding it in a test tube holder. Record your feeling if
there is any odor in table 7.3.
6. Place the test tube in an ice water bath until cool to the touch.
7. Carefully add, dropwise with shaking, 6 M NaOH to the cool solution until basic.
Note: You may need more than 7 mL of base.
8. Hold a piece of moist pH paper over the mouth. Record the pH reading in table 7.3 in addition to
record your feeling if there is any odor.

Report Sheet 1
Grade Group Names:
Date:
Table 7.2: Physical properties of Amines and Amides

Odor Solubility pH
Original
With HCl H2 O Ether HCl H2 O
solution
NH3
Triethylamin
e
Aniline
N,N-
Dimethylanili
ne
Acetamide
Triethylamine and concentrated hydrochloric acid observation:
Table 7.3: Hydrolysis of Acetamide

Any color change of
PH Odor
solution
heating the
solution
After heating ------- -------
Adding NaOH

Report Sheet 2
1. From table 7.2, Compare between the observations of the solubility of adding HCl to each sample
with the observation of adding water to them?
2. Write the structure of the salt that forms when diethylamine, (CH3CH2)2NH, is mixed with
hydrochloric acid.
3. Write the chemical equation for the reaction of triethylamine with concentrated hydrochloric acid.
4. Write the equations that account for what happens in the hydrolysis of the acetamide solution in
(a) acid and in (b) base.
a.
b.

Experiment 8
Carbohydrates
Objectives
1. To become familiar with the reducing or non-reducing nature of carbohydrates.
2. To experience the enzyme-catalyzed and acid-catalyzed hydrolysis of acetal groups.
Introduction
Carbohydrates are polyhydroxy aldehydes, ketones, or compounds that yield polyhydroxy aldehydes
or ketones upon hydrolysis. Rice, potatoes, bread, corn, candy, and fruits are rich in carbohydrates. A
carbohydrate can be classified as a monosaccharide (glucose or fructose); a disaccharide (sucrose or
lactose), which consists of two joined monosaccharide; or a polysaccharide (starch or cellulose),
which consists of thousands of monosaccharide units linked together. Monosaccharide exist mostly as
cyclic structures containing hemiacetal (or hemiketal) groups. These structures in solutions are in
equilibrium with the corresponding open chain structures bearing aldehyde or ketone groups.
Glucose, blood sugar, is an example of a polyhydroxy aldehyde. Fig 8.1:
Figure 8.1: The structure of D-

glucose
Disaccharides and polysaccharides exist as cyclic structures containing functional groups such as
hydroxyl groups, acetal (or ketal), and hemiacetal (or hemiketal). Most of the di-, oligo-, or
polysaccharides have two distinct ends. The one end which has a hemiacetal (or hemiketal) on its
terminal is called the reducing end, and the one which does not contain a hemiacetal (or hemiketal)
terminal is the nonreducing end. The name “reducing” is given because hemiacetals (and to a lesser
extent hemiketals) can reduce an oxidizing agent such as Benedict’s reagent. Fig 8.2 is an example:

Figure 8.2: The structure of maltose, a disaccharide.
Not all disaccharides or polysaccharides contain a reducing end. An example is sucrose, which does
not have a hemiacetal (or hemiketal) group on either of its ends (Fig. 8.3).
Figure 8.3: The structure of sucrose
Polysaccharides, such as amylose or amylopectin, do have a hemiacetal group on one of their

terminal ends, but practically they are nonreducing substances because there is only one reducing
group for every 2,000–10,000 monosaccharidic units. In such a low concentration, the reducing group
does not give a positive test with Benedict’s or Fehling’s reagent.
On the other hand, when a nonreducing disaccharide (sucrose) or a polysaccharide such as amylose is
hydrolyzed the glycosidic linkages (acetal) are broken and reducing ends are created. Hydrolyzed
sucrose (a mixture of D-glucose and D-fructose) will give a positive test with Benedict’s or Fehling’s
reagent as well as hydrolyzed amylose (a mixture of glucose and glucose containing
oligosaccharides). The hydrolysis of sucrose or amylose can be achieved by using a strong acid such
as HCl or with the aid of biological catalysts (enzymes).

Starch can form an intense, brilliant, dark blue-, or violet-colored complex with iodine. The straight
chain component of starch, the amylose, gives a blue color while the branched component, the
amylopectin, yields a purple color. In the presence of iodine, the amylose forms helixes inside of
which the iodine molecules assemble as long polyiodide chains. The helix-forming branches of
amylopectin are much shorter than those of amylose. Therefore, the polyiodide chains are also much
shorter in the amylopectin-iodine complex than in the amylose-iodine complex. The result is a
different color (purple). When starch is hydrolyzed and broken down to small carbohydrate units, the
iodine will not give a dark blue (or purple) color. The iodine test is used in this experiment to indicate
the completion of the hydrolysis.
In this experiment, you will investigate some chemical properties of carbohydrates in terms of their
functional groups.
1. Reducing and non-reducing properties of carbohydrates

A. Aldoses (polyhydroxy aldehydes). All aldoses are reducing sugars because they contain free
aldehyde functional groups. The aldehydes are oxidized by mild oxidizing agents (e.g.,
Benedict’s or Fehling’s reagent) to the corresponding carboxylates. For example,
B. Ketoses (polyhydroxy ketones). All ketoses are reducing sugars because they have a ketone
functional group next to an alcohol functional group. The reactivity of this specific ketone
(also called α-hydroxyketone) is attributed to its ability to form an α-hydroxyaldehyde in basic
media according to the following equilibrium equations:
C. Hemiacetal functional group (potential aldehydes). Carbohydrates with hemiacetal

functional groups can reduce mild oxidizing agents such as Benedict’s reagent because
hemiacetal can easily form aldehydes through the following equilibrium equation:
Sucrose is, on the other hand, a nonreducing sugar because it does not contain a hemiacetal
functional group. Although starch has a hemiacetal functional group at one end of its
molecule, it is, however, considered as a nonreducing sugar because the effect of the
hemiacetal group in a very large starch molecule becomes insignificant to give a positive
Benedict’s test.

2. Hydrolysis of acetal groups. Disaccharides and polysaccharides can be converted into

monosaccharide by hydrolysis. The following is an example:
Materials

Chemicals Supplies
Fehling’s reagen 8 test tubes
2% Glucose 1 big test tube
2% Fructose 600 mL beaker serve as boiling water bath
2% Sucrose Boiling chips in the bath
2% Lactose 10 mL graduated cylinder
2% Starch Well plate
0.01 I2-KI solution (Iodine solution) droppers
3 M H2SO4
3 M NaOH
Saliva (Volunteer from each group)
Procedure
Part I: Reducing or Non-reducing Carbohydrates

1. In 600-mL beaker put about 200 mL of tap water with a few boiling chips and heat it till boiling
and use it as water bath during the experiment.
2. Label 5 clean test tubes as Glucose, Fructose, Sucrose, Lactose, and Starch respectively then add
10 drops of each sample to be tested.
3. Add 2 mL (approximately 40 drops) of Fehling’s solution to each test tube.
4. Place the test tubes in a boiling water bath prepared in step 1 for 5 min.
5. Record your results in table 8.2 in the report sheet. Which of those carbohydrates are reducing
carbohydrates?

Part II: Hydrolysis of Carbohydrates
A. Hydrolysis of sucrose (acid versus base catalysis)
1. Label two test tubes as (no. 1 and no. 2) then and place in each 3 mL of 2% sucrose
2. To the first test tube (no. 1), add 3 mL of water and 3 drops of 3 M H2SO4 solution
3. To the second test tube (no. 2), add 3 mL of water and 3 drops of 3 M NaOH solution
4. Heat the test tubes in a boiling water bath prepared in part I-step 1 for about 5 min.
5. Cool both solutions to room temperature. To the contents of test tube no. 1, add about 10
drops of 3 M NaOH solution or until red litmus paper turns blue.
6. Test a few drops of each of the two solutions (test tube nos. 1 and 2) using the well plate
with Fehling’s reagent. Record the observation color of the solutions in table 8.3 in the
report sheet.
B. Hydrolysis of starch (enzyme versus acid catalysis)
1. Label a new two clean test tubes as (no. 1 and no. 2) then and place in each 2 mL of 2%
starch solution in each of two labeled test tubes.
2. To the first test tube (no. 1), add 2 mL of your own saliva.
Note: Use a 10-mL graduated cylinder to collect your saliva.
3. To the second test tube (no. 2), add 2 mL of 3 M H2SO4 solution.
4. Place both test tubes in a water bath that has been previously heated to 45°C then allow
the test tubes with their contents to stand in the warm water bath for 30 min.
5. Transfer a few drops of each solution into well plate cells and to each sample add 2 drops
of iodine solution. Record the observation color of the solutions in table 8.3 in the report
sheet.

C. Acid catalyzed hydrolysis of starch
1. Place 5 mL of starch solution in a large test tube and add 1 mL of 3 M H2SO4 solution.
Mix it by gently shaking the test tube.
2. Heat the solution in a boiling water bath for about 5 min.
Note: keep tracking the time for every 5 min heating to be able to fill table 8.3.
3. Using a clean dropper, transfer about 3 drops of the heated starch solution into a well plate
and then add 2 drops of iodine solution.
4. If the solution gives turns blue with iodine solution, continue heating by repeating step 2-4
till the solution no longer gives a blue color with iodine solution, stop heating and record
the time needed for the completion of hydrolysis in table 8.3 in the report sheet
Note: Rinse the dropper very thoroughly before each test.

Report Sheet 1
Group Names:
Grade
Date
Table 8.2: Part I: Reducing or Non-reducing Carbohydrates

Substance Reducing or non-reducing carbohydrates
Glucose
Fructose
Sucrose
Lactose
Starch
Table 8.3: Hydrolysis of Carbohydrates

Hydrolysis of sucrose (acid versus base catalysis)
Fehling’s reagent
Sample Condition of hydrolysis
(positive or negative)
1 Acidic (H2SO4)
2 Basic (NaOH)
Hydrolysis of starch (enzyme versus acid catalysis)
Sample Condition of hydrolysis Iodine test (positive or negative)
1 Enzymatic (saliva)
2 Acidic (H2SO4)
Acid catalyzed hydrolysis of starch
Sample Heating time (min.) Iodine test (positive or negative)
1 5
2 10
3 15
4 20

Report Sheet 2
1. Sucrose is a non-reducing sugar. After complete acid hydrolysis, will there be reducing
groups? How many per sucrose molecule?
2. Circle and label the hemiacetal functional group and the acetal functional group in the
following carbohydrates:
3. When a reducing sugar reacts with Fehling’s reagent, what will be the product besides Cu2O?

Experiment 9
Hydrolysis of Starch a Polysaccharide
Objectives
To experience the enzyme-catalyzed and acid-catalyzed hydrolysis of polysaccharides
Introduction
Starch is a complex carbohydrate of high molecular weight having the composition (C6H10O5)n. It
is known to be composed of glucose units, for this is the sole monosaccharide obtained on complete
hydrolysis of the material by means of hydrochloric acid. Under the influence of certain enzymes
hydrolysis proceeds just as easily as with a mineral acid but not as far. The enzyme diastase, which is
present in malt, brings about hydrolysis to maltose. The saliva contains an enzyme, ptyalin, which
likewise converts starch largely into maltose. This disaccharide is susceptible to hydrolysis by
hydrochloric acid and hence cannot be isolated in the reaction of starch with this acid although it
undoubtedly is an intermediate product. It is probable that starch is made up of a series of maltose
units linked together by oxygen bridges by a process akin to a polymerization.
Starch occurs in plants in the form of granules and it is not directly soluble in water. When it is boiled
with water the granules swell and the material gradually passes into solution, possibly as the result of
a partial depolymerization
Even soluble starch must consist of fairly large molecules, for the solution is colloidal. This can be
shown clearly by a simple dialysis experiment. If a solution containing both starch and glucose is
placed in a parchment bag and this is left suspended in pure water for some hours, tests will show that
glucose has diffused through the parchment and that the starch has not.
Starch is easily recognized, and at the same time distinguished from the other carbohydrates, by the
characteristic blue color of some product of unknown nature which is formed by reaction with iodine.
The test is very useful in following the progress of a dialysis or-a hydrolysis. In the latter case an
intermediate stage is reached at which a red color is produced with iodine.
At this point the solution contains a certain amount of substances known as dextrins. These are
probably tetra- or pentasaccharides, but their exact nature is unknown; a mixture of such products ("
dextrin ") is used as a mucilage.

Materials
Chemicals Supplies
Starch Boiling water bath and Ice-water bath
I2/KI solution 1 small test tube
5 mL Saliva (Volunteer from each group) 100 mL Beaker (2) and 250 mL Beaker
Conc. HCl 10 mL Graduated cylinder
Thermometer and glass rod
Well-plate and dropper
* TA must prepare I2/KI solution by dissolving 1 g of iodine in a solution of 2 g KI in 8 mL of H2O
in a beaker.
Procedure
1. Measure 100 mL of distilled water and transfer it to a 250 mL beaker and heat to boiling.
2. Prepare colloidal solution of starch by mixing thoroughly 1 gram of starch with 10 mL of

distilled water in a 100 mL beaker until a uniform paste is obtain
3. Transfer the paste with stirring to the beaker in step 1 to make starch solution.
4. Take 1 mL from solution in step 3 and transfer it to a small test tube, cool it using the ice
water bath then add 1 drop of I2/KI solution.
Note: step 4 is a test to make sure that there is no reducing sugars present prior to the
reaction.
5. Divide the starch solution from step 3 into two parts. Cool the first part to 40°C and add 5 mL
of saliva, stir it well.
6. Record the time and test for starch at 1 minute intervals by transferring a few drops of the
starch solution from the first part and add 1 drop of I2/KI solution.
Note: observe any change in the appearance of the solution and record the time of noting
such change, this will represent the time of a complete hydrolysis
7. To the second part of the starch solution from step 5, add 1 mL of conc. HCl and heat the
solution using the boiling water bath.
8. Make test for starch for every 5 minutes interval by repeating I2/KI test in step 6
Note: observe any change in the appearance of the solution and record the time of noting
such change, this will represent the time of a complete hydrolysis.

Report Sheet 1
Grade Group Names:
Date:
Table 9.2: Hydrolysis of Starch

enzyme catalyzed hydrolysis of starch
Part Condition of hydrolysis Iodine test (positive or negative)

1 Enzymatic (saliva)
Sample Cooling time (min.) Iodine test (positive or negative)
1 1
2 2
3 3
4 4
Acid catalyzed hydrolysis of starch
Part Condition of hydrolysis Iodine test (positive or negative)

2 Acidic (HCl)
Sample Heating time (min.) Iodine test (positive or negative)

1 5
2 10
3 15
4 20

Report Sheet 2
1. The hydrolysis of starch was stopped when the iodine test no longer gave a blue color. Does
this mean that the starch solution was completely hydrolyzed to glucose? Explain.
2. Which hydrolysis of the starch is faster? On the basis of this experiment estimate what will
happen to the digestion of a piece of bread (containing starch) when you chew it thoroughly?

Experiment 10
Isolation and Identification of Casein (Protein)
Objectives
1. To isolate the casein from milk under isoelectric conditions.
2. To perform some chemical tests to identify proteins.
Introduction
Casein is the most important protein in milk. It functions as a storage protein, fulfilling nutritional
requirements. Casein can be isolated from milk by acidification to bring it to its isoelectric point. At
the isoelectric point, the number of positive charges on a protein equals the number of negative
charges. Proteins are least soluble in water at their isoelectric points because they tend to aggregate
by electrostatic interaction. The positive end of one protein molecule attracts the negative end of
another protein molecule, and the aggregates precipitate out of solution.
On the other hand, if a protein molecule has a net positive charge (at low pH or acidic condition) or a
net negative charge (at high pH or basic condition), its solubility in water is increased.
In the first part of this experiment, you are going to isolate casein from milk which has a pH of about
7. Casein will be separated as an insoluble precipitate by acidification of the milk to its isoelectric
point (pH = 4.6). The fat that precipitates along with casein can be removed by dissolving it in
alcohol.
In the second part of this experiment, you are going to prove that the precipitated milk product is a
protein.

The identification will be achieved by performing a few important chemical tests.

1. The biuret test. This is one of the most general tests for proteins. When a protein reacts with
copper(II) sulfate, a positive test is the formation of a copper complex which has a violet color.
This test works for any protein or compound that contains two or more of the following groups:
2. The ninhydrin test. Amino acids with a free -NH2 group and proteins containing free amino
groups react with ninhydrin to give a purple-blue complex.

3. Heavy metal ions test. Heavy metal ions precipitate proteins from solution. The ions that are most
commonly used for protein precipitation are Zn2+, Fe3+, Cu2+, Sb3+, Ag+, Cd2+, and Pb2+. Among
these metal ions, Hg2+, Cd2+, and Pb2+ are known for their notorious toxicity to humans. They can
cause serious damage to proteins (especially enzymes) by denaturing them. This can result in
death. The precipitation occurs because proteins become cross-linked by heavy metals as shown
below:
Victims swallowing Hg2+ or Cd2+ ions are often treated with an antidote of a food rich in proteins,
which can combine with mercury or lead ions in the victim’s stomach and, hopefully, prevent
absorption! Milk and raw egg white are used most often. The insoluble complexes are then
immediately removed from the stomach by an emetic.
4. The xanthoprotein test. This is a characteristic reaction of proteins that contain phenyl rings
Concentrated nitric acid reacts with the phenyl ring to give a yellow-colored aromatic nitro
compound. Addition of alkali at this point will deepen the color to orange.
The yellow stains on the skin caused by nitric acid are the result of the xanthoprotein reaction.

Materials

Chemicals Supplies
Liquid milk 250 mL Erlenmeyer flask
Glacial acetic acid 600 mL beaker serve as boiling water bath
Ethanol Thermometer and stirring rod
1:1 mixture of diethyl ether-ethanol Cheese cloth
2% glycine Rubber band
2% gelatin 100 mL beaker (2)
2% albumin 5 small test tubes
1% Tyrosine Hot plate
10% NaOH
1 M CuSO4
Ninhydrin reagent
Pb(NO3)2
Hg(NO3)2
NaNO3
Conc HNO3
Procedure
Part I: Isolation of Casein

1. Heat in a 600-mL beaker about 200 mL of tap water till boiling.
2. Put a 250-mL Erlenmeyer flask on the scale, tare it, and weigh 50 g of milk.
3. Immerse the Erlenmeyer flask in the water bath you prepared in step 1 and Stir the solution
constantly with a stirring rod. See Fig. 10.1.
Figure 10.1: Precipitation of Casein

4. When the bath temperature has reached about 40°C, remove the flask from the water bath, and
add about 10 drops of glacial acetic acid while stirring. Observe the formation of a precipitate.
5. Filter the mixture into a 100-mL beaker by pouring it through a cheese cloth which is fastened
with a rubber band over the mouth of the beaker (Fig. 10.2).
Figure 10.2: Filtration of Casein
6. Remove most of the water from the precipitate by squeezing the cloth gently.
7. Discard the filtrate in the beaker. Using a spatula, scrape the precipitate from the cheese cloth into
the empty clean beaker.
8. Add 25 mL of ethanol to the beaker in step 7. After stirring the mixture for 5 min., allow the solid
to settle.
9. Carefully decant the liquid that contains fats into another beaker to discard this liquid.
10. To the residue (solid), add 25 mL of a 1:1 mixture of diethyl ether-ethanol. After stirring the
resulting mixture for 5 min., collect the solid by vacuum filtration, refer to figure 5.1.
11. Spread the casein on a paper towel and let it dry.
12. Weigh the dried casein and calculate the percentage of casein in the milk. Record it in the report
sheet.

Part II: Chemical Analysis of Proteins
A. The biuret test
1. Label 4 clean test tubes as glycine, gelatin, albumin, and Tyrosine respectively then add 15
drops of each sample to be tested.
2. In test tube no. 5 put a small amount of Casein prepared in PartI and add 15 drops of distilled
water.
To each of the test tubes, add 5 drops of 10% NaOH solution and 2 drops of 1M CuSO4
solution with stirring. Record your observation in table 10.2 in the report sheet.
Note: Observing a purplish-violet color is an evidence of the presence of proteins.
B. The ninhydrin test
1. Repeat steps 1 and 2 from part II (A).
2. To each of the test tubes, add 5 drops of ninhydrin reagent
3. Heat the test tubes in a boiling water bath you prepared in part I (step 1) for about 5 min and
record your observation in table 10.2 in the report sheet.
C. Heavy metal ions test
1. Label 3 clean test tubes as no.1, no. 2, and no. 3.
2. Add 2 mL of liquid milk in each test tube.
3. Add a few drops of each of the following metal ions to the corresponding test tubes as
indicated below and record your observation in table 10.2 in the report sheet
a. Pb2+ as Pb(NO3)2 to test tube no.1
b. Hg2+ as Hg(NO3)2 to test tube no.2
c. Na+ as NaNO3 to test tube no.3
D. The xanthoprotein test.
1. Repeat steps 1 and 2 from part II (A).
2. To each of the test tubes, add 10 drops of concentrated HNO3 while swirling.
3. Heat the test tubes carefully in a warm water bath. Observe any change in color and record
your observation in table 10.2 in the report sheet

Report Sheet 1
Group Names:
Grade
Date
Part I: Isolation of Casein

1. Weight of milk ______________ g
2. Weight of dried casein ______________ g
3.
Percent of casein = ______________ %
Table 10.2 Part II: Chemical analysis of proteins

C. Heavy metal D. Xanthoprotein
A. Biuret test B. Ninhydrin test
ion test test
Samples
Color formed after Precipitates Color formed before
Color formed
heating formed or after heating
2% glycine
2% gelatin
2% albumin
casein + H2O
1% tyrosine
A. Which of these chemicals gives a positive test with Biuret reagents? --------------------------
B. Which of these chemicals gives a positive test with Ninhydrin reagents? ---------------------
C. Which of these metal ions gives a positive test with casein in milk? -------------------------
D. Which of these chemicals gives a positive test with Xanthoprotein reagent? ----------------

Report Sheet 2
1. Casein has an isoelectric point at pH 4.6. What kind of charges will be on the casein in its native
environment, that is, in milk?
2. Would the amino acid, glycine, give a positive biuret test? Explain
3. What are the three most toxic heavy metal ions?

Experiment 11
Analysis of Lipids
Objectives
To investigate the lipid composition of common foods such as corn oil, butter, and egg yolk.
Introduction
Lipids are chemically heterogeneous mixtures. The only common property they have is their
insolubility in water. We can test for the presence of various lipids by analyzing their chemical
constituents. Foods contain a variety of lipids, most important among them are fats, complex lipids,
and steroids. Fats are triglycerides, esters of fatty acids and glycerol.
Complex lipids also contain fatty acids, but their alcohol may be either glycerol or sphingosine. They
also contain other constituents such as phosphate, choline, or ethanolamine or mono- to oligo-
saccharides. An important representative of this group is lecithin, a glycerophospholipid, containing
fatty acids, glycerol, phosphate, and choline.
The most important steroid in foods is cholesterol. Different foods contain different proportions of
these three groups of lipids.
Structurally, cholesterol contains the steroid nucleus that is the common core of all steroids.
There is a special colorimetric test, the Lieberman-Burchard reaction, which uses acetic anhydride
and sulfuric acid as reagents, that gives a characteristic green color in the presence of cholesterol.
This color is due to the -OH group of cholesterol and the unsaturation found in the adjacent fused
ring. The color change is gradual: first it appears as a pink coloration, changing later to lilac, and
finally to deep green.

When lecithin is hydrolyzed in acidic medium, both the fatty acid ester bonds and the phosphate ester
bonds are broken and free fatty acids and inorganic phosphate are released. Using a molybdate test,
we can detect the presence of phosphate in the hydrolysate by the appearance of a purple color.
Although this test is not specific for lecithin (other phosphate containing lipids will give a positive
molybdate test), it differentiates clearly between fat and cholesterol on the one hand (negative test),
and phospholipid on the other (positive test).
A second test that differentiates between cholesterol and lecithin is the acrolein reaction. When lipids
containing glycerol are heated in the presence of potassium hydrogen sulfate, the glycerol is
dehydrated, forming acrolein, which has an unpleasant odor. Further heating results in polymerization
of acrolein, which is indicated by the slight blackening of the reaction mixture. Both the pungent
smell and the black color indicate the presence of glycerol, and thereby fat and/or lecithin.
Cholesterol gives a negative acrolein test.

Materials
Chemicals Supplies
0.2 g pure cholesterol 6 large test tubes
0.2 g pure glycerol 6 small test tubes
0.2 g lecithin 250 mL beaker serve as boiling water bath
Corn oil 100 mL beaker (2)
Butter Cheese cloth
Egg yolk 250 mL Erlenmeyer flask
6 M HNO3 Hot plate
6 M NaOH
Molybdate solution
Ascorbic acid
KHSO4
Sucrose
Chloroform
Acetic anhydride
Conc. H2SO4
Procedure
Part I: Phosphate Test

1. Prepare a water bath by boiling about 100 mL of tap water in a 250-mL beaker on a hot plate.
2. Label 6 clean test tubes as cholesterol, glycerol, lecithin, Corn oil, Butter and Egg yolk
respectively then add 0.2 g of the sample to each test tube to be tested.
3. Hydrolyze the compounds by adding 3 mL of 6 M nitric acid to each test tube
4. Place the test tubes in the boiling water bath for 5 min. after that leave them to cool down.
Note: Do not inhale the vapors.
5. Neutralize the acid by adding 3 mL of 6 M NaOH. Mix. During the hydrolysis, a precipitate may
form, especially in the egg yolk sample.
6. The samples in which a precipitate appeared must be filtered. Place a piece of cheese cloth on top
of a 100-mL beaker. Pour the turbid hydrolysate in the test tube on the cheese cloth and filter it.
Figure 10.2
7. Transfer 2 mL of each neutralized (and filtered) sample into small clean and labeled test tubes.
Add 3 mL of a molybdate solution to each test tube and mix the contents.

8. Heat the test tubes in a boiling water bath for 5 min. cool them to room temperature.
9. Add 0.5 mL of an ascorbic acid solution and mix the contents thoroughly.
10. Wait 20 min. for the development of the purple color. Record your observations in table 11.2 in
the report sheet.
Part II: The Acrolein Test for Glycerol
1. Label 7 clean test tubes as cholesterol, glycerol, lecithin, Corn oil, Butter, Egg yolk and sucrose
respectively then add 0.1 g of the sample to each test tube to be tested.
2. Add 1 g of KHSO4, in each of seven test tubes
Note: It is important that step 3 test be performed under the hood because of the pungent odor
of the acrolein.
3. Set up your Bunsen burner in the hood and gently heat each test tube, one at a time, over the
Bunsen burner flame, shaking it continuously from side to side. When the mixture melts it slightly
blackens, and you will notice the evolution of fumes. Stop the heating. A pungent odor,
resembling burnt hamburgers, is the positive test for Glycerol. Sucrose also will be dehydrated
and will give a black color. However, its smell is different, and thus is not a positive test for
acrolein.
Note1: Smell the test tubes by moving them sideways under your nose or waft the vapors. Do
not inhale the fumes directly.
Note2: Do not overheat the test tubes, for the residue will become hard, making it difficult to
clean the test tubes. Record your observations on the Report Sheet.
Part III: Lieberman-Burchard Test for Cholesterol
1. Repeat step 1 in part II
Note: The next step should be done in
2. Add 3 mL of chloroform and 1 mL of acetic anhydride to each test tube.
3. Add 1 drop of conc. H2SO4 to each mixture. Mix the contents and record the color changes, if
any. Wait 5 min. Record again the color of your solutions in table 11.2 in the report sheet.

Report Sheet 1
Group Names:
Grade
Date
Table 11.2: Record observation
Cholestero
Tests Lecithin Glycerol Corn oil Butter Egg yol Sucrose
l
1. Phosphate
a. color
b. Conclusions
2. Acrolein
a. Odor
b. Color
c. Conclusions
3. Lieberman-
Burchard
a. Initial color
b. Color after 5
min.
c. Conclusion

Report Sheet 2
1. Based on the intensity of color developed in your test for cholesterol, which food contained the
most and which contained the least cholesterol?
2. Cholesterol in tissues is sometimes esterified by fatty acids. (a) Draw the structure of cholesterol
oleate. (b) Would this ester give a positive Lieberman-Burchard test?
3. Why was it necessary to hydrolyze the samples with nitric acid before performing the molybdate
test?
4. Why must you wear gloves in performing the phosphate test?

6. References
 Frederick A. Bettelheim, Joseph Landesberg (2007), Manual for Laboratory Experiments for
General, Organic, and Biochemistry, 4th Ed, Saunders College Pub.
 Louis Fieser (1941). Manual for Experiments in Organic Chemistry, 2nd Ed, Harvard University,
D. C. Heath and Company.
 (2011, June), Laboratory Manual for Organic Chemistry I, Al-Akhawayn University, School of
Science & Engineering
 (2009, Fall), Chem 21: Nucleophilic Substitution, Amherst College, Department of Chemistry
https://www.amherst.edu/system/files/media/0931/Exp7.pdf
 Elaine N., Melissa H., Jack R., (2015), Organic Chemistry with Vernier Lab book, Experiment
13: SN1: Synthesis of tert-Butyl Chloride, Vernier Lab Safety Instructions Disclaimer
http://www.vernier.com/products/books/chem-o/


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Komar University of Science and

Preface and Acknowledgments

In preparing this laboratory manual, I am thankful to the efforts of KUST

I anticipate that students will find it inspiring in studying different aspects of

KOMAR UNIVERSITY OF SCIENCE AND TECHNOLOGY (KUST) 1

KOMAR UNIVERSITY OF SCIENCE AND TECHNOLOGY (KUST) 2

1. Common Laboratory Apparatus*

Items Name Picture Items Name Picture

Volumetric Flask Test Tubes

Watch Glass Crucible and lid

Dropper Evaporating dish

Funnel Graduated cylinder

KOMAR UNIVERSITY OF SCIENCE AND TECHNOLOGY (KUST) 1

Items Name Picture Items Name Picture

Mortar and pestle Beaker

Stirring rod Erlenmeyer Flask

Thermometer Bunsen burner

KOMAR UNIVERSITY OF SCIENCE AND TECHNOLOGY (KUST) 2

Items Name Picture Items Name Picture

Wire gauze Clamp

Iron ring Clay triangle

Wash bottle Safety goggles

Corks Rubber Stopper

Test tube holder Spatulas

Suction Filteration Well plate

KOMAR UNIVERSITY OF SCIENCE AND TECHNOLOGY (KUST) 3

Items Name Picture Items Name Picture

Water bath Electronic Lab Scale

Hot plate pH-meter

KOMAR UNIVERSITY OF SCIENCE AND TECHNOLOGY (KUST) 4

2. Chemistry Lab Safety

2.1 Laboratory Attire

4. Long hair must be tied back.

KOMAR UNIVERSITY OF SCIENCE AND TECHNOLOGY (KUST) 5

2.2 Laboratory Conduct

2. No unauthorized experiments are to be performed. If you are curious about trying a

4. Always wash your hands before leaving lab.

6. Clean any spill near the balance immediately and

7. Notify the instructor immediately in case of an

8. Consider all chemicals to be hazardous as a result

9. Know what chemicals you are using. Carefully read

KOMAR UNIVERSITY OF SCIENCE AND TECHNOLOGY (KUST) 6

15. Dispose of chemicals properly. Waste

16. Clean up all broken glassware immediately

18. Beware of hot glass--it looks exactly like cold glass.

2.3 Fire Case

1. In the event of fire, do not panic.

KOMAR UNIVERSITY OF SCIENCE AND TECHNOLOGY (KUST) 7

3. Writing Lab Reports

4. Submission of Lab Reports

KOMAR UNIVERSITY OF SCIENCE AND TECHNOLOGY (KUST) 8

KOMAR UNIVERSITY OF SCIENCE AND TECHNOLOGY (KUST) 9

1. Synthesize tert-butyl chloride via an SN1 reaction.

Figure 1.1: Conversion of t-butanol into t-butyl chloride by an SN1 mechanism

KOMAR UNIVERSITY OF SCIENCE AND TECHNOLOGY (KUST) 10

Part I: Synthesis of tert-butyl chloride

1. Prepare an ice-water bath in a 250 mL beaker.

2. Measure 15 mL of concentrated HCl using a graduated cylinder and carefully transfer to a 50 mL

4. Set up the reaction in a Separatory funnel. Figure 1.2:

Unknown no. . The unknown compound is .