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ORGANIC ANALYTICAL

CHEMISTRY
Laboratory Guide

This module provides guidelines for safety, Laboratory


Report and note book. It’s also providing students with
Laboratory manual for subject of Organic Analytical
Chemistry (CLB 10803).

FIRST EDITION
2010
Laboratory Information

Before each lab session, you should prepare by reading the lab manual, reference book and
summarized it in a jotter book. We expect you to have a good understanding of the purpose,
details of the procedure, the use of all chemicals and any significant hazards, and the underlying
science of the experiment when you come to lab.
CONTENTS

Preface i

Laboratory safety Guidelines ii

Safety Declaration Form vi

Chemistry Laboratory Report Guidelines vii

Laboratory Notes Books Guidelines x

Experiment

1 Saponification Reaction reaction of Fat: Soap Production 1

2 Analysis of Food Colour 4

3 Determination of Benzoic acid /caffeine in Soft drink 8

4 Organic Synthesis: Formation of Ester. 11

REFERENCES 14

APPENDICES 15
PREFACE

This manual provides laboratory guidelines, safety declaration form, Chemistry Lab
Report guidelines and Laboratory manual for subject of Organic Analytical Chemistry
(CLB 10803).

The primary purpose of this manual is to compile all necessary information regarding
laboratory component in one manual.

The manual contains four parts. Part 1 provides a description of laboratory quidelines and
safety declaration form. It is compulsory for student to understand all those guidelines
and submit safety declaration for recording purposes. Part 2 is laboratory report
guidelines containing all requirements such as format and arrangement in order to
produce good quality of laboratory report. Part 3 is guidelines for preparation of
laboratory notes book. Part 4 is compilation of laboratory manual that will provide
student practical guidelines in Organic Analytical Chemistry.

There may be shortcomings which we had overlooked but hopefully these should not
hinder the process of enhancing laboratory skill.

Mohd Zulkhairi Abdul Rahim


Nor Nadiah Mohamad Yusof
PART 1

LABORATORY SAFETY GUIDELINES

General Guidelines

1. Conduct yourself in a responsible manner at all times in the laboratory.


2. Be familiar with your lab assignment before you come to the lab. Follow all
written and verbal instructions carefully. If you do not understand a direction or
part of a procedure, ask the instructor before proceeding.
3. No student may work in laboratory alone. The lab instructor or co-coordinator
grant exceptions on a case by case basis.
4. When first entering a laboratory, do not touch any equipment, chemicals or other
materials in the laboratory area until you are instructed to do so.
5. Do not eat, drink beverages or chew gum in the laboratory. Do not use laboratory
glassware as containers for food or beverages.
6. Smoking is not allowed in any indoor area.
7. No music allowed in the laboratory. Radio (including walkman) and other
entertainment devices are not permitted.
8. No cellular phone is allowed in this laboratory.
9. Perform only those experiments authorized by the instructor. Never do anything
in the laboratory that is not called for the laboratory procedures or by your
instructor. Carefully follow all instructions, both written and oral. Unauthorized
experiments are prohibited.
10. Observe good housekeeping practices. Work areas should be kept clean and tidy
at all times.
11. Horseplay, practical jokes, and pranks are dangerous and prohibited.
12. Always work in a well-ventilated area.
13. Bring only your laboratory instructions, worksheets and report to the work area.
Other materials (books, purses, backpacks, etc) should be stored in the cabinet.
14. Know the locations and operation procedures of all safety equipment including
the first aid kit, eyewash station, safety shower, spill kit and fire extinguisher.
15. Be alert and proceed with caution at all times in the laboratory. Notify the
instructor immediately of any unsafe condition you observe.
16. Label and equipment instructions must be read carefully before use. Set up and
use the prescribed apparatus as directed in the laboratory instructions provided by
your instructor.
17. Experiments must be personally monitored at all times. You will be assigned a
laboratory station at which to work. Do not wander around the room, distract
other students or interfere with laboratory experiments or others.
18. Write your name and equipment use every time you come in to the laboratory in
the log book.
19. Defeating safety devices or using equipment in a manner other than that which is
intended will be grounds for dismissal from the lab.
Clothing
1. Safety goggles and safety jacket must be worn whenever you work in lab.
2. Gloves should be worn whenever you use chemicals that cause skin irritations or
need to handle hot equipment.
3. Mask should be worn every time you prepare the chemicals.
4. Safety shoes and hard hat should be worn at all times while in the laboratory.
5. Contact lenses should not be worn in the laboratory unless you have permission
from your instructor.
6. Dress properly during a laboratory activity.
7. Long hair, dangling jewelry and loose or baggy clothing are a hazard in the
laboratory. Long hair must be tied back and dangling jewelry and loose or baggy
clothing must be secured.
8. Sandal, open-toed shoes, high heels or shoes with holes in the sols will not be
worn in the lab.
9. Short and skirts are not permitted.
10. Instructor and laboratory assistant have a right dismiss to you from the laboratory
if they found that you are not wearing proper safety clothing.

Handling Chemicals
1. Treat chemicals with respect and understand the chemicals you are using with
Material Safety Data Sheet (MSDS). The MSDS are available in the analytical
room.
2. All chemicals in the laboratory are to be considered dangerous. Do not touch,
taste or smell any chemical unless specifically instructed to do so.
3. Check the label on chemical bottles before removing any of the contents. Take
only much chemical are you need. Smaller amounts often work better than larger
amounts.
4. Label all containers and massing papers holding dry chemicals.
5. Never return unused chemicals to their original containers.
6. Never use mouth suction to fill a pipette. Use pipette bulb or pipette filler.
7. Acids must be handled with extreme care. Always add acids slowly to water, with
slow stirring and swirling, being careful of the heat produced, particularly with
sulfuric acid.
8. Handle flammable hazardous liquid over a pan to contain spills. Never dispense
flammable liquids anywhere near a flame or source of heat.
9. Never take chemicals or other materials from the laboratory area.
10. Take good care when transferring acids and other chemicals from one part of the
laboratory to another. Hold them securely and in the method demonstrated by the
instructor as you walk.
11. All wastes generated during the course of an experiment must be disposed of
according to the lab instructor’s directions.
12. Never mix chemicals in sink drains.
13. Sinks are to be used only for water and those solutions designated by the
instructor.
14. Solid chemicals, metals, matches, filter paper, and all other insoluble materials are
to be disposed of in the proper waste containers, not in the sink.
15. Checks the label of all waste containers twice before adding your chemicals waste
to the container.
16. Cracked or broken glass should be placed in the special container for “broken
glass”.
17. Keep hands away from your face, eyes, mouth and body while using chemicals.
Wash your hands with soap and water after performing all experiments.

Personal Hygiene
1. Wash hands before leaving the lab and before eating.
2. Gloves should be removed before leaving the lab, using telephones, or entering
common areas
Accidents and Injuries
1. Report any accidents (spill, breakage, etc) or injury (cut, burn, etc) to the
instructor immediately, no matter how trivial it may appear.
2. If you or your lab partners are hurt, immediately tell to the instructor.
3. If a chemical should splash in your eye(s), immediately flush with running water
from the eyewash station for at least 20 minutes. Notify the instructor
immediately.
4. Spills should be cleaned up immediately.

Handling Glassware and Equipment


1. Inserting and removing glass tubing from rubber stopper can be dangerous.
Always lubricate glassware (tubing, thistle tubes, thermometer, etc) before
attempting to insert it in a stopper. Always protect your hands with tower or
cotton gloves when inserting glass tubing into, or removing it from a rubber
stopper.
2. When removing an electrical plug from its socket, grasp the plug, not the
electrical cord.
3. Hands must be completely dry before touching an electrical switch, plug or outlet.
4. Examine glassware before each use. Never use chipped or cracked glassware.
5. Never use dirty glassware.
6. Do not immerse hot glassware in cold water; it may shatter.
7. Report damaged electrical equipment immediately. Look for things such as frayed
cords, exposed wires and loose connections. Do not use damaged electrical
equipment.
8. If you do not understand how to use a piece of equipment, ask the instructor for
help.
9. Be careful when lifting heavy objects. Lift comfortably, avoid unnecessary
bending, twisting, reaching out, and excessive weights, lift gradually and keep in
good physical shape.
10. Do not transfer a glassware form one laboratory to another without permission
from instructor.
Heating Substances

1. Do not operate a hot plate by yourself. Take care that hair, clothing, and hands
are a safe distance from the hot plate at all times. Use of hot plate is only allowed
in the presence of the teacher.
2. Heated glassware remains very hot for a long time. They should be set aside in a
designated place to cool, and picked up with caution. Use tongs or heat protective
gloves if necessary.
3. Never look into a container that is being heated.
4. Do not place hot apparatus directly on the laboratory desk. Always use an
insulated pad. Allow plenty of time for hot apparatus to cool before touching it.
5. If leaving a lab unattended, turn off all ignition sources and lock the doors.

Ended the Experiments


9. At the end of the laboratory sessions, you should;
⇒ Shut-off main gas outlet
⇒ Turn-off the water inlet
⇒ Desk top, floor area and sink are clean
⇒ All equipment is cool, clean and arranged
9. All equipment use should be flushed using deionized water.
SAFETY DECLARATION FORM

The Dean/Head of Campus


Universiti Kuala Lumpur
Malaysian Institute of Chemical and Bioengineering Technology
Lot 1988, Vendor City Industrial Area
Taboh Naning, 78000 Alor Gajah
Malacca

Dear Sir,

SAFETY DECLARATION

I ………………………………..………………………………………………………….
ID No ………………………. declare that I have read and understood the safety rules and
regulations in UniKL MICET. I hereby agree to abide by all the rules and regulations
stated in the safety guidelines.

9. I hereby understood the contents and will disciplinary action will be taken against
me, if I do not abide by the stated rules.

9. I am fully responsible for all my actions during laboratory sessions.

Thank you.

Yours faithfully,

……………………………….
Name:
Matrix No:
Subject:
Date:
PART 2

CHEMISTRY LAB REPORT FORMAT

You should type your lab report. Make sure that you check your document for any spelling
errors. Each lab report is worth 100 points. You should also read the student handbook on the
subject of plagiarism. Your data and observations will be similar, but your interpretations
should not be written identically. You may not copy another student’s lab report in part or in its
entirety. If you are found guilty of this infraction, you and the person from whom you copied
will both lose points or shared total marks. In extreme cases or repeated offenses, both students
may receive a zero for the lab.

Title

Use a separate title page. Include the title of the experiment, YOUR NAME, and the date.
Also clearly indicate the name(s) of your lab partner(s).

Summary/Abstract (not more than 1 page)

It should be written after conclusion of experiment OR project. Its cover briefly about
Introduction, Objectives, Methodology, Result & discussion, Conclusion

Objectives

State the objectives of the experiment or report (point form)

Example:

The objective of the experiments was …….

Introduction

Background to the work / experiment

Example:

The purpose of this experiment was to identify the present of heavy metal in river water by
using Atomic Absorption Spectroscopy (AAS). AAS is ………….

Theory / formula used in the experiment. Do not just copy word for word from the lab handout.
The introduction should be of 1– 3 pages.
Materials

List the chemicals and equipment needed to perform the experiment.

Procedure

Write the procedure in chronological order. Again, DO NOT COPY DIRECTLY from the lab
handout. Rearrange your procedure become a passive sentence.

Example:

Prepare 0.5M NaOH solution (from manual)


0.5M NaOH was prepared (your report)

Result & Discussion

Analyze all data qualitative and quantitative. Then transfer finding into Table, Graph,
Histogram and Pie chart if necessary. This includes any observations. Make sure that your
graphs have titles, labeled axes with units, and legends. You should include the proper units
with any numbers, as well as use the proper number of significant figures based upon the lab
equipment used. DO NOT place any calculations or data analysis in this section. It may be a
good idea to reproduce here any data tables that you completed during the lab. Base on above
point, discuss on your findings and relate to your theory and objective of experiment.

Example:

Table 1: X vs. Y

Samples X (unit) Y (unit)


A
B
C
D
50
45 y = 4.4557x + 0.9714
40 R2 = 0.9893

parameter X (unit)
35
30
25
20
15
10
5
0
0 2 4 6 8 10 12
parameter Y (unit)

Figure 1: Relationship between X and Y

Conclusions

This is the most important section. Please include the summary of the results and relate in brief
the findings / results with the theory. Answer the questions, “What did you learn?”, “Did I
accomplish the purpose?”, “How would I improve the experiment next time?”.
Recommendation is optional. The conclusion should be one paragraph of 5 – 7 sentences.

References

Write down any sources such as your textbook, the Internet, electronic
encyclopedia, books, etc. that you used.

• A list of lab manuals, books, reports, journal, world wide web (www) etc.
• Arrangement (year, alphabetical order)
• Author, title, publisher, year, chapter or page number

Example:

Smith J.M and Van Hess H.C., Introduction to Chemical Engineering Thermodynamics,
McGraw-Hill, New York, 2001, p229

Appendix

Here is where you attach any material that you think is pertinent to the lab report such as
summary of calculation involved. Also answer any questions here that are in the lab report.
You do not have to re-write the questions, but label and number them appropriately.
PART 3

LABORATORY NOTEBOOKS

You are required to use a bound notebook in CLD 10004 Lab to record all primary data and
observations. You should prepare your notebook before coming to a lab by writing the
title of the experiment on a new numbered page, summarizing relevant information from
the lab manual, and starting calculations involving molar masses, etc. Take note of
theoretical ideas and special instructions given by your instructor at the start of each
experiment. Your notebook should be a complete record of your work in lab. Notes should be
able to understand in the future, not just during the current experiment. Good note taking in a
lab is a valuable skill that you can learn with a little effort and practice.

Guidelines to be followed:
1. Always bring your notebook with you to lab. You will be graded on the completeness of
your previous note taking and your preparation for the current experiment. You may use
your notebook during a lab quiz.
2. Number the pages sequentially and reserve space at the beginning for a table of contents.
3. Take your notebook during laboratory hours and record all values directly in it – not on
loose scraps of paper.
4. Specify each measured quantity by name and include the units.
5. If you make a mistake in your notebook, simply draw a solid line through the error and write
the correction nearby.
6. Tables greatly simplify data entry; they should be set up before coming to lab.
7. Write down all observations if necessary – don’t rely on your memory.
8. Save time by doing trial calculations in your notebook before filling out any report sheets.
9. Save time by making preliminary sketches of graphs (flow Chart) on the ruled lines in your
notebook.
PART 4

EXPERIMENT 1

SAPONIFICATION REACTION OF FAT: SOAP PRODUCTION

OBJECTIVE

• To synthesize a sample of hard soap


• To test the soap produced

INTRODUCTION

The procedure of making soap involves the basic hydrolysis (saponification) of a fat.
Chemically, fats are referred to as triglycerides. They contain ester functional groups.
Saponification involves heating fat with an alkaline solution. The alkaline solution hydrolyzes
the fat to alcohol and the salt of a long chain carboxylic acid (soap).When common salt is
added, the soap precipitates. The soap is washed free of unreacted alkaline solution and molded
into bars.

O HO CH2
O
R C O CH 2

O + 3 NaOH 3R C O Na +
R C O CH HO CH
Sodium salt of an acid (soap)
O

R C O CH 2

Fat HO CH2
glycerol
MATERIAL AND METHODS

Materials

Chemicals:

NaOH 50% water/ethanol mixture NaCI Solution


95% ethanol 4 % calcium shloride solution
Fat Trisodium phospahte

Apparatus:
Conical Flasks Hirsch / Buchner funnel
Beaker Watch glass
Filter funnel

METHODS

Preparation of Soap

1. Prepare a NaOH solution (about 0.25 g sodium hydroxide dissolved in a mixture of 1.0
ml of distilled water and 1.0 ml of 95% ethanol)..
2. Place about 0.25 g of fat in a 50 ml conical flask and add the prepared sodium hydroxide
solution to the flask.
3. Heat the mixture in a in a bath of 100 oC.
4. Cover the flask with some aluminum foil to help reduce evaporation. Swirl the
Erlenmeyer flask every few minutes. Use tong to do this.
5. The soap will precipitate from the boiling mixture within 20 minutes.
6. If you observe that some alcohol and water is evaporating from the flask, you may
add up to 0.4 ml of a 50 % water/alcohol mixture to replace the solvent.
7. Heat the mixture for a maximum time of 25 minutes.
8. Place 4 ml of NaCI solution in a 15 ml beaker and transfer the saponified mixture from
flask to beaker.
9. Stir the mixture while cooling the beaker in an ice-water bath.
10. Collect the prepared soap on a Hirsch funnel of ice cold distilled water to remove excess
NaOH.
11. Continue to draw air through the filter for a few minutes to partially dry the product. Test
your soap with the procedure below.

Analysis of Data

1. Remove about 0.01 g of soap from the filter paper and placed it in a clean 10 ml
graduated cylinder
2. Add 3 ml of distilled water, close the cylinder with your thumb and shake the mixture
vigorously for about 15 sec. After about 30 sec standing, Record your observation.
Note down the level of the foam.
3. Add 5 – 10 drops of 4% calcium chloride solution to the soap mixture from a Pasteur
pipette .Shake the mixture for 15 sec and allow it to stand for 30 seconds. Record your
observation on the effect of addition the calcium chloride.
4. Then add 0.5 g of trisodium phosphate and shake the mixture again for 15 seconds. After
30 sec. Again observe and record the result.

APPENDIX

Pre Laboratory Question

1. Give a definition of saponification


2. Explain how soap can function as “dirt remover”.
3. Synthetic detergent functions in the same way as soaps. Give the advantages of
synthetic detergent over soaps.

Post Laboratory Question

1. Reaction of fat with NaOH will produced long chain carboxylic acid (soap) in form of
Bar. What would be happen if sodium Hydroxide (NaOH) is replaced by Potassium
hydroxide (KOH).
EXPERIMENT 2

ANALYSIS OF FOOD COLOUR

OBJECTIVE

(1) To determine λmax of Colourant (wavelength scan)


(2) To prepare a serial dilution and generate a standard calibration graph for sample
quantitation (photometric scan)

INTRODUCTION

Food coloring (colouring) is any substance that is added to food or drink to change its color. .
Synthetic Food Colours, also known as Artificial Food Colours, are manufactured chemically
and are the most commonly used dyes in the food, pharmaceutical and cosmetic industries.
Besides that , a growing number of natural food dyes are being commercially produced, partly
due to consumer concerns surrounding synthetic dyes. Some examples include Caramel
coloring (E150), made from caramelized sugar, used in cola products and also in
cosmetics.Annatto (E160b), a reddish-orange dye made from the seed of the Achiote A green
dye made from chlorella algae (chlorophyll, E140) and etc.

All those colourant can be analyze by using an ultraviolet/visible spectrophotometer. Figure


1.0 below gives a schematic diagram of a double beam spectrophotometer. Instruments for
measuring the absorption of U.V. or visible radiation are made up of the following
components; Sources (UV and visible), Wavelength selector (monochromator), Sample
containers, Detector, Signal processor and readout

Figure 11.1 Schematic diagram of double beam UV – Vis spectrophotometer


When a beam of parallel radiation passes through a layer of solution of thickness, b (cm) and a
concentration, C (moles per liter) of an absorbing species, absorption of radiation occurs. The
transmittance (T) of the solution is the fraction of incident radiation transmitted by the solution.
Transmittance is often expressed as a percentage. The absorbance (A) of a solution is defined
as the negative log of the transmittance (T) of the solution. The absorbance is directly
proportional to the path length of the radiation through the solution and the concentration of the
absorbing species.

A=εbc

Where; A = absorbance (no unit)

ε = molar absorptivity coefficient (M-1 cm-1)

b = pathlength (cm)

c = concentration (M or mol/L)

This relationship between absorbance, A and ε bc is known as Beer’s Law. Beer’s Law is
successful in describing the absorption behavior of dilute solutions only. At high
concentrations, absorbance of the solution does not obey Beer’s Law and is no longer
proportional to C.

In this experiment you are going to conduct two main applications which are wavelength
scanning and photometric scanning.
MATERIALS AND METHODS

Materials

Chemicals

100 ppm colorant stock (100 ml), Unknown (two), Distilled water

Apparatus

Perkin – Elmer UV/Vis Spectrophotometer Lambda EZ210, Sample cuvettes, path length 1
cm, Volumetric flask 50 ml (five) , Pipette 5 ml, 10 ml and 25 ml (one each),Rubber bulb
(three),Beaker 100 ml (one),Graduated cylinder 50 ml (one),Dropper (one),Labeling
sticker,Tissue paper

Methods

1. Prepare serial dilutions (5 ppm, 15 ppm, 25 ppm, 35 ppm and 45 ppm) in 50 ml


volumetric flask from the 100 ppm carmoisine stock.
2. After preparing the serial dilutions, your instructor will brief on the standard
operating procedure of Perkin – Elmer UV/Vis Spectrophotometer Lambda EZ210.
3. Fill a cuvette with 45 ppm dilution and another cuvette with blank solution; insert
them in the sample compartment. Wipe clean the sides of the cuvettes and
remember not to touch on the clear surface. Do the wavelengths scan and obtain
the λmax. Record your data..
4. For photometric scan, fill the cuvette as step no 3 but use the serial dilution
prepared and scan one by one. Record the absorbance readings and look at the
standard calibration graph produced.
5. Also determine the concentrations of Unknown #1 and Unknown #2.
Analysis of Data

The purpose of wavelength scan is to determine at what wavelength the carmoisine able to
absorb in the range of 200 nm to 700 nm. From spectrum obtain, please identify λmax .

The purpose of photometric scan is to determine the concentration (single component) of an


unknown sample, after generating a working 'standard curve' from a series of known standards
(known concentration). Record the absorbance readings for a series of prepared dilution
generate standards calibration curve and identify concentration of unknowns. All
calculation must show in detail.

Pre Laboratory Question

1. State the Beer’s lambert law.

2. What is the volume needed to prepare a 50 ppm of carmoisine from a 100 ppm of
carmoisine in 100 ml volumetric flask?

Post Laboratory Question

1. Why we need to wipe the sides of the cuvette clear surface?

2. Describe the function of wavelength scanning and photometric scanning.


EXPERIMENT 3

Determination of Benzioc acid / Caffeine in Soft drink

Objective :

1. To Identify the present of Benzoic acid/ Caffeine in soft drink sample


2. To determine amount of caffeine in soft drink sample.

Introduction:

High Performance Liquid Chromatography (HPLC) is a chemistry based tool for quantifying
and analyzing mixtures of chemical compounds. It can be used to separate compounds that are
dissolved in solution. HPLC instruments consist of a reservoir of mobile phase, a pump, an
injector, a separation column, and a detector. Compounds are separated by injecting a plug of
the sample mixture onto the column. The different components in the mixture pass through the
column at different rates due to differences in their partitioning behavior between the mobile
liquid phase and the stationary phase.

The area of this peak (in relation to the area of other peaks) is proportional to the concentration
of that particular species in the sample. The identity can also be found by comparing the
sample peaks to standards. Identical substances (peaks) will have identical retention times.
MATERIALS AND METHODS

Materials:

Isocratic HPLC system with UV detector, C18 column, Vacuum, Funnel, 0.45 µm filter paper
0.45 µm filter syringe,100 µL syringe,60 mL syringe,Volumetric flask

Chemicals

Caffeine 1000 ppm standard (stock solution), Methanol (HPLC grade), Double distilled water
(filtered with 0.45 µm filter paper), Soft drink sample

Methods:

1. Preparation of Benzoic acid/ caffeine standards


⇒ Prepare standard caffeine samples of 20 ppm, 40 ppm, 60 ppm, 80 ppm and 100 ppm
by diluting portions of the 1000 ppm solution with distilled water.
2. Preparation of soda samples
⇒ Obtain a soft drink sample.
⇒ Degas the sample by placing it in a vacuum flask and connecting the flask to a vacuum
pump or water aspirator. Leave it under vacuum until no more bubbles appear in the
soda sample. (If no vacuum is available, allow the soda to stand open overnight.)
⇒ Filter the degassed soda through #42 filter paper.

3. After preparing the serial dilution and sample, your instructor will brief on standard
operating procedure of HPLC.

Analysis of data

1. Use standard benzoic acid /caffeine retention time to identify the benzoic
acid/caffeine peak and then record their retention time.
2. By using above information, clarify and justify the present of benzoic acid/caffeine in
the soda sample.
3. Record different concentration of standards peaks area and construct a standard
calibration curve (concentration vs peak area).
4. Measure the caffeine peak in the soda sample chromatograph, and use standard
calibration curve (concentration vs peak area) to determine the concentration of
Benzoic acid/ caffeine in the soda sample.
Note: All raw data must be record in table form.

Pre – Lab Questions:

1. Briefly explain how HPLC is used as a separation technique.


2. What is the purpose of the mobile phase? Of the stationary phase?
3. What is the purpose of the caffeine standards?

Post – Lab Questions:

1. Why does the syringe have to be carefully rinsed before each use?
2. Retention of caffeine in standards. How could you be identify a peak in the soda was
caffeine and not another substance by using retention time?
EXPERIMENT 4

ORGANIC SYNTHESIS: FORMATION OF ESTER

OBJECTIVE

• To synthesis ethyl acetate (ethyl ethanoate)

INTRODUCTION

Chemist use organic synthesis to make larger amounts of useful natural compounds and to
invent totally new compounds. Depending on the choice of R’ and R, we have a variety of the
final ester, R’COOR. Small chain side groups give very aromatic compound, while long –
chain side groups form waxy compounds.

Carboxylic acid alcohol ester water


The reaction of a carboxylic acid with an alcohol to produce an ester plus water is known as
the Fisher esterification reaction. A mineral acid, usually sulfuric acid is used as a catalyst.

MATERIALS AND METHODS

Chemical:

Ethanol, Glacial acetic acid, Conc. sulfuric acid, 30% Sodium carbonate solution, Calcium
chloride, Granular anhydrous Calcium chloride, anti-bumping granules.

Apparatus:
Round bottom flask, Water condenser, retort stand, separating funnel
Methods

1. Set – up a reflux system.


2. Mix 50 ml of 95% ethanol and 50 ml of glacial acetic acid thoroughly in a 250 ml round –
bottomed flask. Add slowly with cooling and shaking 10 ml of concentrated H2SO4. Ensure
that the mixture is homogenous, and then fit the flask with a reflux water – condenser and
boil the mixture gently for 10 minutes. Cool the flask and its content.
3. Rearrange the position of the condenser for distillation set –up.
4. Put a few boiling chips in the flask. A filter flask, whose side arm is joined to a rubber tube
leading over the edge of laboratory bench, is used as a receiver. Ethyl acetate is highly
flammable. Therefore any vapors should be conducted off the table towards the floor.
Distlled off about 2/3 of the mixture.
5. Transfer the distillate to a separating funnel and add about 25 ml of 30 % Na2CO3
solution. Stopper the funnel, invert it and shake it, opening the stopcock from time to time.
Allow the two layers to separate. Carefully run off and reject the lower layer, ensuring
that the sodium carbonate is removed as completely as possible.
6. Prepare a solution of 25 g of calcium chloride in 25 ml of water. Add if to crude ethyl
acetate in the funnel. Shake vigorously. Allow the mixture to separate. Run off the lower
aqueous as completely as possible.
7. Run the ethyl acetate into a small conical flask. Add a few lumps of granular anhydrous
calcium chloride. Shake occasionally until the liquid is clear.
8. Decant the liquid some anti bumping granules. Arrange for distillation (with a 0 – 100 oC
thermometer in the apparatus). Pre – weight the receiving flask. The distilling flask
should be placed in cold water bath, which is gradually heated.
9. The ether that is always formed in this reaction will distill off at 35 – 45 oC and may be
discarded. Continue to heat and collect the fraction that boils between 74 oC and 79 oC.
10. Weight your product and calculate the percentage yield.
11. Run FTIR of your product and characterize it smell.
12. Inject substrate and product into Gas chromatography.
Analysis

1. From mass of acid, determine the percent yield of your final product
(Show all calculation in detail)
2. Interpret the FTIR of this compound. Identify the principles peaks.
3. Interpret Gas chromatography result.

Appendix

Pre – Lab Question

1. Reaction of acetic acid with ethanol to produce will produce ethyl ethanoate and water.
Based on that the reaction, how many grams of ethyl ethanoate would be produced if 50
ml of ethanol were react with 50 ml acetic acid? (given ethanol: 0.8 g/ml and acetic acid :
1.06 g/ml and ethyl acetate: 0.9g/ml). Calculate the percentage yield if 50.0 g of ethyl
ethanoate was obtained from the experiment.

Post – Lab Question


1. What are percent yields? How this can be improves?
2. Based on result above, does the FTIR show any contaminant from initial reactants?
Explain.
REFERENCE

1. Francis A Carey, Organic Chemistry, 7th Edition, McGraw Hill


ISBN : 0073311847 / 9780073311845
2. T.W. Graham Solomon,Organic Chemistry, 8th Edition, Wiley QD253.2.S65
2004
3. Douglas A. Skoog, Donald M. West and F. James Holler, Fundamentals of
Analytical Chemistry, 8th ed., Saunders College Publishing, 1997.
QD75.22.F86 2003
4. Mohan, Jag, Organic Analytical Chemistry: Theory and Practice, Alpha Science
International, Ltd, 2004
ISBN :0849339529 / 9780849339523
5. Mohan,Jag, Organic Spectroscopy: Principles and Applications, Alpha Science
International, Ltd.
ISBN :0849339529 / 9780849339523
APPENDIXS

Mass Force
1 lbm = 0.453592 kg 1 lbf = 4.448222 N
1 ton = 2000 lbm 1N = 0.224809 lbf = 1 kg.m/s2
1 kg = 2.20462 lbm

Volume Density
1 ft3 = 0.028317 m3 1kg /m3 = 0.062428 lbm/ ft3
1 L = 0.001 m3
1 m3 = 35.32 ft3
1 cm3 = 0.06102 in3
1 gal =0.0037854 m3
1 gal/min = 6.31 x 105 m3/s

Pressure

1 pascal (Pa) = 1 N/m2


1 atm = 760 mmHg = 760 torr
1 atm = 101.325 KPa
Table 1.0 Characteristic Infrared Absorption frequencies

Bond Compound Type Frequency range, cm-1


2960-2850(s) stretch
Alkanes
1470-1350(v) scissoring and bending
C-H
1380(m-w) - Doublet - isopropyl, t-
CH3 Umbrella Deformation
butyl
3080-3020(m) stretch
C-H Alkenes
1000-675(s) bend
Aromatic Rings 3100-3000(m) stretch
C-H Phenyl Ring Substitution Bands 870-675(s) bend
Phenyl Ring Substitution Overtones 2000-1600(w) - fingerprint region
3333-3267(s) stretch
C-H Alkynes
700-610(b) bend
C=C Alkenes 1680-1640(m,w)) stretch
C C Alkynes 2260-2100(w,sh) stretch
C=C Aromatic Rings 1600, 1500(w) stretch
C-O Alcohols, Ethers, Carboxylic acids, Esters 1260-1000(s) stretch
Aldehydes, Ketones, Carboxylic acids,
C=O 1760-1670(s) stretch
Esters
Monomeric -- Alcohols, Phenols 3640-3160(s,br) stretch
O-H Hydrogen-bonded -- Alcohols, Phenols 3600-3200(b) stretch
Carboxylic acids 3000-2500(b) stretch
3500-3300(m) stretch
N-H Amines
1650-1580 (m) bend
C-N Amines 1340-1020(m) stretch
C N Nitriles 2260-2220(v) stretch
1660-1500(s) asymmetrical stretch
NO2 Nitro Compounds
1390-1260(s) symmetrical stretch

v - variable, m - medium, s - strong, br - broad, w - weak


∼THIS IS THE END OF THE LABORATORY MANUAL ~