BSK 1432

LABORATORY MANUAL

EXPERIMENTS IN
ANALYTICAL CHEMISTRY

WAN NORFAZILAH WAN ISMAIL

NURLIN ABU SAMAH
NAZIKUSSABAH ZAHARUDIN

FACULTY OF INDUSTRIAL SCIENCES &

TECHNOLOGY
 i

TABLE OF CONTENTS

Table of Contents i

Introduction ii

Jotter Book iv

Report and Format vi

Assessment of Report and Jotter Book xi

Safety Rules xii

EXPERIMENT 1: Basic Laboratory Techniques 1

EXPERIMENT 2: Calibration of a Pipette 7

EXPERIMENT 3: Quantitative Analysis of Dye in Water Sample 11

EXPERIMENT 4: Colorimetric Determination of Phosphate 17

EXPERIMENT 5: Determination of Monoprotic and Diprotic Acid – Acid-Base Titration 21

EXPERIMENT 6: Determination of Chloride by the Mohr Method – Precipitation Titration 27

EXPERIMENT 7: Determination of Amino Acids – Potentiometric Titration 30

EXPERIMENT 8: Determination of Ascorbic Acid in Vitamin C – Redox Titration 34

Appendices 39-40
 ii

INTRODUCTION

This manual is prepared for BSK 1432 Analytical Chemistry Laboratory and includes
the experiments, which are related to the topics covered in the BSK 1153 Analytical
Chemistry course. The main purpose of this laboratory is to provide the students an
appreciation for the potential applications and limitations of analytical methods of
analysis. It is also aimed to provide the students an opportunity to develop their
abilities in the laboratory skills required for accurate and precise chemical analyses.
Therefore, it is expected that with an acceptable degree of proficiency the students
will grasp and apply the principles of analytical chemistry to obtain reliable results in
the laboratory experiments.

The fundamental assumption made by students as they enter this course is that
students are an adult person with a scientific curiosity ready to learn analytical
chemistry. This may not be a valid assumption in all cases, but if this is not students
approach to this course, then try to pretend that it is. There is much work to be done.
This may be one of the most demanding courses students will take. However, it is
also one of the most useful courses for anyone planning to do chemistry at beginning
level.

This course is ultimately practical to all aspects of scientific careers. The lab skills
which students will learn and refine are the most "marketable" part of students’
degree. They will have hands on experience to all but a small fraction of the
apparatus, instruments and methods used in the various aspects of chemistry. BSK
1432 will introduce the most commonly used methods and basic instruments to
prepare students for work and research in chemical laboratories. So, enjoy it!

Attendance is required and you are expected to attend all scheduled laboratory
sessions. If you miss more than two-lab sessions without a valid reason you will be
automatically dropped from lab. Should you be sick and cannot come to lab, you
need to bring a doctor’s excuse from the UNIVERSITY HEALTH CENTER or an
official report from a government hospital. Also, if you cannot be available for an
official reason and you know this ahead of time, you should notify the teaching
 iii

assistants in advance of the lab period and make sure you obtain the required
documentation. Remember that this is group work and if you cannot be there, you
must inform the other members of your group. Be on time since your group members
will depend on you to start the experiments. Tardiness will be noted by the teaching
assistant and can affect your grade.

Note:
This document is used only as a compulsory lab manual for this course. Students are
advice to refer to other related materials.
 iv

JOTTER BOOK

Orderly record keeping is essential in a laboratory. A well-kept notebook makes

writing lab reports much easier. It also helps when trying to troubleshoot problems
that may arise during an experiment. In an industrial setting a laboratory notebook is
a legal record of research activities. Inventions and ideas that are not properly
documented may not hold up in patent litigation. A portion of your grade will be
based on your jotter book. Student MUST have ONE jotter and some points to note
are given below:

Front Page: Please write FULL NAME, ID NUMBER, COURSE CODE & NAME,
and SEMESTER & SESSION (Figure 1).

BSK1432
ANALYTICAL CHEMISTRY LABORATORY

Name: _____________________________
ID No.: _____________________________
Semester/Session: ____________________

Figure 1: Example of front cover of jotter book.

 v

First Page: Refer to the Table 1. Full table is given in APPENDIX A. Please print
and put in your jotter.

The jotter must be in A4 size booklet with hard cover. Students may write the
keyword / key point of the experiment for each pre-lab session. Students are required
to bring their jotter to the lab and NOT the lab manual. For those who FAIL to do
that, their marks will be deducted. The jotter will be checked by the lab instructor
every week before the class starts and lab instructor will sign on the first page of
jotter for submission of the jotter and lab report (Table 1).

Table 1: First Page (Signature Page) of jotter book. Lab instructor will sign for both
jotter & Lab Report submission.

Title Jotter Lab Report

Experiment 1 Lab instructor please Lab instructor please sign here

sign here

Experiment 2 Lab instructor please Lab instructor please sign here

sign here

The jotter book is not meant to be a picture perfect record of what you did. Many
students take 'notes' on scrap paper with the intent of transferring the data to their
jotter books later. This practice should be avoided at all times. Copies of the relevant
spread sheets will be turned in along with your report. A duplicating of the spread
sheets is preferable to photocopies and keeps them in jotter book.
 vi

COMPREHENSIVE LABORATORY REPORTS & FORMAT

Lab reports are the written synthesis of the work that you performed in the
laboratory. An outsider skilled in chemistry should be able to read your report and
understand what you did, why you did it and what you discovered. You will find that
good written and verbal communications skills are keys to a successful career in the
chemical sciences.

As with all scientific reports, it should be written in past tense (since you presumably
are writing the report after you did the work) and with passive voice. Grammatically,
passive voice is a bit awkward, however it lends objectivity to the report. For
example, one may choose to write “I found the weight percent to be 0.59%”. It is not
important that you as an individual made the measurement. Properly phrased one
would simply write “The weight percentage was found to be 0.59%”. Avoid the use
of words such as “I” or “we”, the experimental result should not depend on the
experimentalist. LABEL your figures and tables. Keep in mind that the reader may
not be familiar with your experiment.

For comprehensive laboratory reports, students will be given 1 WEEK to prepare

and SUBMIT before they start the next experiment. If they FAIL to submit on time,
their marks will be deducted. Students MUST WRITE from the abstract until
references by TYPE WRITING. Since the experiment will be done in grouping, the
lab instructor will allow the students to share their reports for the front page, title,
objective, chemicals/apparatus/instruments, methodology, finding & observation
EXCEPT abstract, interpretation of data, conclusion and references. For those who
do the PLAGIARISM for the abstract, interpretation of data, conclusion and
references, they will automatically get ZERO for that parts. No second chances
allowed.
 vii

The format for the comprehensive laboratory report as follows:

Writing Format
Paper size: A4
Margin: 2.5 inch from left, right, top and bottom.
Font type: Times New Roman
Font size: 12
Spacing: Single spacing for abstract and 1.5 spacing for other contents.

Front page
Please use the separately given sample of cover page.

Abstract
One (1) page is for abstract. In this page, you must summarize your report including
objective of the experiment, description of the precise problem, method used, results
and conclusion. Abstract must be short and clear. Abstract writing is the last session
after all the contents are completed. Please refer the example of abstract given in
APPENDIX B.

Title

Introduction
Make an introduction about the experiment. Divide into 2 subtopics:
1.1 Introduction (literature review and theory)
Make a literature about the topic that related to your experiment. You can
also put the theory or principle occurred during process or reaction.
Furthermore, try to discuss and make a comparison from the previous
studies related to the experiment. You MUST put the reference (name of
author, year) at the end of the statement you cited from any resources in
your text.
1.2 Objective
 viii

Chemicals/Apparatus/Equipment
List all the chemicals, equipment and apparatus that you used throughout the
experiment. Then, explain the purpose and procedure for each equipment used
briefly.

Methodology
This section describes the physical work that you performed in the lab. Since the
procedures are already written for you it is not necessary to re-write them in detail.
You should briefly write the entire procedure in FLOW DIAGRAM.

Results/Finding & Observation

This section contains, like the title suggests, your results and that’s it. What it does
not contain is an in depth discussion of what the results mean. You should include
tables of raw data where appropriate (see Table 2 below), calibration plots and
figures (spectra, diagrams etc…). The results section is where you will report
analytical results with confidence intervals and values you determined such as rate
constants. If the experiment analyzes a value by multiple methods, break the results
into subsections according to analyte then method. A sentence such as “The weight
percent of KHP in sample 1 – 3 was found to be 25.28 ± 0.13 at 95% confidence by
titration with standard NaOH.” should be at the end of each subsection when
appropriate. It is sometimes useful to put your results into a table. This makes it easy
for the reader (or grader) to find important values. Tables always contain a caption
above the table itself. Grid lines should be limited to separations between headers
(and footers) and the data. The table below illustrates some of these ideas. If you are
unsure of how to format a data table consult a style guide or refer to journals for
examples. An example table is shown on the following page.

Table 2: Sodium hydroxide standardization results.

Mass KHP (g) Volume of NaOH (mL) [NaOH] (mol L-1)
Trial 1 0.5426 28.04 0.09476
Trial 2 0.6137 31.58 0.09514
Trial 3 0.5973 30.63 0.09547
Trial 4 0.5011 25.83 0.09497
Trial 5 0.4973 25.16 0.09679
Final Result (± 95% C.I.) [NaOH] = 0.0954 ± 0.0001
 ix

Figures are an efficient way to present numerical data in an easy to visualize format.
It is important to format your plots so that the data is clearly and easily identified.
The choice of fonts, symbols and sizes of these features should be well proportioned.
Use your best judgement here. As with tables, reference to a style guide or journals
for examples is a good way to learn how to do this. All figures should be captioned
below the figure.

0.8
0.7
0.6
Absorbance (Au)

0.5
0.4
0.3
0.2
0.1
0
0 2 4 6 8 10 12

Glucose concentration (mM)

Figure 2: Calibration curve for glucose determined by UV.

Discussion/Interpretation of Data
This section is a chance to comment on the experiment as a whole. In other words,
what do the results signify? What conclusions or predictions can be made based on
your results. If you obtained inconsistent or anomalous results this is your chance to
attempt to explain why. Do your results agree with literature values? Try to relate
them with the theory and past study.

Conclusion
This section mainly consists of two parts: conclusion and future work. First part,
make a conclusion of the experiment and suggest some recommendation to improve
the experiment. Secondly, purpose others experimental work that can also be used
with the same equipment used during this experiment.
 x

References (at least FIVE (5) references)

Every statement or information that cited from any resources should be mentioned in
the text in the form of (Author name, year). Then, at the end of the report list down
all the references that has been cited in the text based on the format below:

(a) Book
Author (Year). Title. Edition (if not the first). Placed published: Publisher.
Example:
Theusen, G. J. and Fabrycky, W. J. (1984). Engineering Economy. 6th ed.
Englewood Cliffs, N. J.: Prentice-Hall.

(b) Journal articles

Author (Year). Title of the article. Title of the Journal. Volume (Number):
page.
Example:
Billings. S. A. (1980). Identification of Nonlinear Systems: A survey. Proc.
IEE, Part D. 127(6): 272-284.

Similarity Percentage (Turnitin)

All reports must be submitted to Turnitin system (assign by coordinator) in order to
check for similarity percentage. This is to avoid any plagiarism. Acceptable
similarity percentage allowed is < 25%. More than the allowable percentage will be
given penalty 10% from the total report marks.

OTHERS:
1) No limit page for the report but minimum page is 5 pages.
2) Please see your lecturer if you do not clear about the format. Report submitted
without following the format will be rejected.
3) All report must be submitted in hard copy and by typewritten.
4) The report will be assessed based on the format, content and the interview/
question conduct throughout the lab experiment.
 xi

ASSESSMENT OF REPORT & JOTTER BOOK

Comprehensive Laboratory Report and Jotter Percentage & Marks

Assessment Percentage (%)

Abstract 5
Introduction & Objectives 10
Materials & Methods 10
Results 15
Calculations 10
Discussion 20
Conclusions 5
References 10
Grammar and spelling 5
Organization & format 5
Appendices 5
Total 100

Overall Marks for BSK 1432

Assessment Percentage (%)

Lab Report (× 8) 60
Observation 10
Practical Test 30
Total 100
 xii

SAFETY RULES

The chemistry laboratory is not a dangerous place to work as long as all necessary
precautions are taken seriously. In the following paragraphs, those important
precautions are described. Everyone who works and performs experiments in a
laboratory must follow these safety rules at all times. Students who do not obey the
safety rules will not be allowed to enter and do any type of work in the laboratory
and they will be counted as absent. It is the student’s responsibility to read carefully
all the safety rules before the first meeting of the lab.

Eye Protection: Because the eyes are particularly susceptible to permanent damage
by corrosive chemicals as well as flying objects, safety goggles must be worn at all
times in the laboratory. Prescription glasses are not recommended since they do not
provide a proper side protection. No sunglasses are allowed in the laboratory.
Contact lenses have potential hazard because the chemical vapours dissolve in the
liquids covering the eye and concentrate behind the lenses. If you have to wear
contact lenses consult with your instructor. If possible try to wear a prescription
glasses under your safety goggles. In case of any accident that a chemical splashes
near your eyes, immediately wash your eyes with lots of water and inform your
instructor. Especially, when heating a test tube do not point it towards anyone.
Always assume that you are the only safe worker in the lab. Work defensively. Never
assume that everyone else as safe as you are. Be alert for other’s mistakes.

Cuts and Burns: Remember you will be working in a chemistry laboratory and
many of the equipment you will be using are made of glass and it is breakable.
Lubricate both the tubing and the hole in the stopper with water when inserting glass
tubing or thermometers into stoppers. Handle tubing with a piece of towel and push it
with a twisting motion. Be very careful when using mercury thermometer. It can be
broken easily and may result in mercury contamination. Mercury vapour is an
extremely toxic. When you heat a piece of glass it gets hot very quickly and
unfortunately hot glass look just like a cold one. Handle glass with tongs. Do not use
any cracked or broken glass equipment. It may ruin an experiment and worse, it may
cause serious injury. Place any broken glass in the proper waste glass container. Do
not throw them into the wastepaper container or regular waste container.

Poisonous Chemicals: All of the chemicals have some degree of health hazard.
Never taste any chemicals in the laboratory unless specifically directed to do so.
Avoid breathing toxic vapours. When working with volatile chemicals and strong
acids and bases use ventilating hoods. If you are asked to smell the odours of a
substance do so by wafting a bit of the vapour toward your nose. Do not stick your
nose in and inhale vapour directly from the test tube. Always wash your hands before
leaving the laboratory. Eating and drinking any type of food are prohibited in the
laboratory at all times. Smoking is not allowed. Anyone who refuses to do so will be
forced to leave the laboratory.

Clothing and Footwear: Everyone must wear a lab coat during the lab and no shorts
and sandals are allowed. Students who come to lab without proper clothing and shoes
will be asked to go back to change his or her clothing. If they do not come on time
 xiii

they will be counted as an absent. Long hair should be securely tied back to avoid the
risk of being set on fire. If large amounts of chemicals are spilled on your body,
immediately remove the contaminated clothing and use the safety shower if
available. Make sure to inform your instructor about the problem. Do not leave your
coats and back packs on the benches because they may be contaminated. No
headphones, mp3 players or cell phones are allowed in the lab because they interfere
with your ability to hear what is going on in the Lab. Cell phones must be turned off.

Fire: In case of fire or an accident, inform your instructor at once. Note the location
of fire extinguishers and, if available, safety showers and safety blankets as soon as
you enter the laboratory so that you may use them if needed. Never perform an
unauthorized experiment in the laboratory. Never assume that it is not necessary to
inform your instructor for small accidents. Notify him/her no matter how slight it is.

Laboratory Care and Waste Disposal

Remember that the equipment you use in this laboratory will also be used by many
other students. Please leave the equipment and all workspaces as you wish to find
them. After the end of the each lab, clean off your work area. Wash your glassware.
When weighing any material on the balances, do not weigh directly onto the balance
pan. Weigh your material on a piece of weighing paper. The balances are very
sensitive instruments and should be treated with great care.

If you take more reagents than you need, do not put excess back into the bottle. It
may be contaminated. Threat it as waste and dispose of it accordingly. It is most
likely that, during any experiment you will perform, you will generate some waste
chemicals and solutions to dispose of. Never put them down the sink unless
specifically told to do so by your instructor. There will be inorganic, organic, and
solid waste containers in the lab. Dispose of your waste in the appropriate container.
 1

EXPERIMENT 1
Basic Laboratory Techniques

INTRODUCTION
The purpose of this experiment is to introduce several of the tools and techniques
necessary for success in this course. The Analytical Chemistry laboratory requires a
substantially higher level of precision, accuracy, laboratory technique, and
cleanliness.

OBJECTIVES:
Part A: To determine the average and median values, the standard deviation, and the
relative standard deviation in the masses of 20 cent coins.
Part B: To provide experience in the correct use of the volumetric flask.
Part C: To provide experience in the correct use and proper reading of burette.

EQUIPMENT/APPARATUS
20 cent coins Stirring rod
Analytical balance Funnel
Tweezer Erlenmeyer flask 250 mL
Volumetric flask 25 mL Gloves
Beaker 50 mL Burette

REAGENTS/CHEMICALS
Aluminium slug
NaOH pellet

PROCEDURE
Part A
In this experiment, you will obtain the masses of five 20 cent coins – first by
weighing each coin individually, and then by weighing all five coins at once,
removing one coin at a time, and obtaining the individual masses of the coins by
difference. The pair of masses determined for a particular coin should agree to within
 2

a few tenths of a milligram. From the data, you will determine the average and
median values, the standard deviation, and the relative standard deviation in the
masses of the coins. You will then weigh an “unknown” aluminium slug, and report
its mass and code number.

After the Lab Assistant has instructed you in the proper use of the electronic balance
and you have become familiar in its use, obtain a set of 20 cent coins, an unknown
aluminium slug, and a pair of tweezers from the Lab Assistant. Never handle the
coins or the slug with your fingers, always use the tweezers.

1. Go the analytical balance that has been assigned to you. Zero it carefully.
Select five 20 cent coins at random from the vial containing the coins.
Keeping track of which coin is which, weigh each coin, one at a time on your
balance. Enter the masses on the data sheet provided.
2. Re-zero the balance. Place all five of the same five coins on the balance pan,
obtain the total mass and enter it on the same data sheet. Remove one coin
from the balance, obtain the mass of the remaining four and record the mass.
Repeat this process, removing one coin at a time. Obtain the individual
weights by subtraction. This process is known as weighing by difference,
which is the way almost all weighing are done in the Analytical Laboratory.
3. Now weigh the unknown aluminium slug and record its mass. Repeat three
times and obtain the average mass.
4. Perform the calculations requested.

Part B
The following experiment is designed to provide experience in the correct use of the
volumetric flask.

1. Tap a very small amount of sodium hydroxide pellet, NaOH, from the stock
bottle onto a piece of folded glassine paper or into a small clean beaker or a
plastic weighing boat. (Note: Chemicals should never be placed back into
stock bottles as this may contaminate the entire bottle. Avoid putting a spatula
into a stock bottle. Tap out a small amount if at all possible.)
 3

2. Tare a clean, dry 50 mL beaker on an electric balance. Add about 0.1 g of

NaOH to the beaker.
NOTE: NEVER transfer chemicals inside an analytical balance.
3. Dissolve the NaOH in about 10 mL of distilled water and stirring gently.
4. Quantitatively transfer the solution into a 25 mL volumetric flask using a
small funnel. To prevent the solution from running down the outside of the
beaker, pour the solution down the stirring rod, and then touch the rod to the
spout of the beaker to remove the last drop. Add more water to the beaker,
stir, and repeat the procedure.
Note the amount of washing required to quantitatively transfer the target
chemical from the beaker to the flask. Finally, rinse the last portions of
solution from the stirring rod into the volumetric flask with a stream of water
from the wash bottle. Rinse the funnel and remove it. Carefully dilute the
solution in the flask until the bottom of the meniscus is even with the
graduation mark.
5. Stopper, invert, and shake the flask. Return it to the upright position, and
allow the air bubble to return all the way to the top of the neck. Repeat until
the solution is completely homogeneous; about 10 inversions and shakings
are required.

Part C
Reading Burette Sections

1. Obtain a set of five “burette sections” from the Lab Assistant. The sections are
normally stored upside down so that the inside surface of the part of the
section where you will take your reading will remain wetted and drain well.
2. Invert each section and tap the section lightly to remove any solvent that
might remain in the sealed tip and let it drain for at least 20-30 sec. Record the
number and reading of each burette section on the form provided. Use a
burette reading card to estimate the readings to the nearest 0.01 mL.
[A burette card is easily made by making a very heavy, black horizontal
rectangle in the center of a 3 x 5” file card.]
3. Repeat for a second set of burette sections.
 4

Use and Proper Reading of Burette

1. Over-fill the burette with distilled water. Make sure that there is no air
bubbles trapped in the tip. If so, rapidly open and close the valve several times
until the bubbles are flushed out.
2. Drain the burette down to someplace between 0 and 1 mL. Do not try to bring
the burette to exactly 0.00 mL, as this is a waste of time. The “zero” reading
on all but an automatic burette is always some finite non-zero reading.
3. Wait at least 20-30 seconds before taking the initial or “zero” reading. Take
the initial or “zero” reading and all other burette readings using a burette
reading card.
4. Now let about 5 mL run into a 250 mL Erlenmeyer flask. Wait at least 30
seconds and take the “final reading”. (The amount of solution in the
Erlenmeyer flask is equal to the final reading minus the “zero” reading.) Write
down the final reading on the form provided, and then ask Lab Assistant to
take the final reading. Compare the two readings.
5. Refill the burette, and take a new zero reading. Now add 30 drops to the
Erlenmeyer flask, and take the final reading. Calculate the average volume of
one drop, then repeat this with 40 drops and calculate the average volume of a
drop. Record these results and compare them.
 5

Spread Sheet for Experiment 1

Part A
Item Coin 1 Coin 2 Coin 3 Coin 4 Coin 5 Total mass
Code
Mass

n
Mean
Median
Standard
deviation
RSD

Item 5 coins 4 coins 3 coins 2 coins 1 coin

Code
Mass
Item Coin 1 Coin 2 Coin 3 Coin 4 Coin 5 Total mass
Individual
mass by
different

n
Mean
Median
Standard
deviation
RSD

Aluminium slug – Code:

1 2 3 Average
Mass

Part B

Item Details
Mass of NaOH pellet
Molar mass of NaOH
No. of mol of NaOH
(Show the calculation)

Volume of solution
Molarity of NaOH solution
(Show the calculation)
 6

PART C
Item Section 1 Section 2 Section 3 Section 4 Section 5
Reading

Item Reading
Initial or “zero” reading
Final reading (after 5 mL less)
Final reading by Lab Assistant

Item Reading
Initial or “zero” reading
Final reading (after 30 drops)
Volume
Average volume of one drop

Initial or “zero” reading

Final reading (after 40 drops)
Volume
Average volume of one drop
 7

EXPERIMENT 2
Calibration of a Pipette

INTRODUCTION
An ordinary laboratory pipette may be expected to deliver its nominal volume with
good precision and good accuracy if it is used in the way recommended. The
calibration of an analytical transfer pipette is a relatively straightforward procedure.
The proper technique is readily learned with practice, care, and attention to detail.
Note that this is a manual technique. It must be learned; it does not magically appear
on the first try. With some practice, however, you ought to be able to handle a pipette
well.

Correction for Buoyancy

A buoyancy error will affect data if the density of the object being weighed differs
significantly from that of the standard weights (inside the balance for an electric
balance). This error is due to the difference in the buoyant force exerted by the
medium (air) on the object weighed and on the weights themselves. The correction
is made using Equation 1:

mcorrected = mobserved + mobserved [(dair/dobject) – (dair/dweights)] [1]

where m is the mass in grams of the corrected and initially observed values of the
object weighed (in this case, distilled water), and d is the density in g/cm3 of air, the
object, and the weights.

For all but the most exacting work, this correction is negligible for solids and liquids
having a density of 2 g/cm3 or greater. Correction for buoyancy is required only for
measurements that require the highest accuracy, for gases, or for low-density solids
and liquids. The density of the weights used in electronic single-pan balances ranges
from 7.8 to 8.4 g/cm3, depending on the composition of the weights. Using dweights =
8 g/cm3 is adequate for most purposes. The dair = 0.0012 g/cm3 at room temperature
and pressure.
 8

OBJECTIVES:
i. to investigate the precision and accuracy of selected pipette.
ii. to illustrate the correction for buoyancy.

EQUIPMENTS/APPARATUS
Pipette bulb Beaker 500 mL
Pipette 10 mL Thermometer
Erlenmeyer flask 50 mL Weighing balance

PROCEDURE
1. Obtain the following equipment: Pipette bulb; 50 mL Erlenmeyer flask with a
solid, dry cork or rubber stopper; 500 mL beaker filled with 400 mL of distilled
water equilibrated to room temperature; and a thermometer.
2. Clean a 10 mL pipette, and ask the Lab Assistant to verify that your pipette is
clean. Cleaning usually accomplished by drawing some warm soapy water into
the pipette bulb, wetting the sides, and “shaking the soap” inside the pipette. If
that doesn’t work, soak the entire pipette in soapy water overnight or longer.
You must clean pipettes, burettes, and other pieces of volumetric glassware, so
that no droplets of reagent remain on the internal surfaces when they are
drained. This is very important for accurate and reproducible results. If
reagent gets “hung up” inside a pipette, you obviously cannot deliver the
nominally stated volume. You should have already cleaned your pipette
properly during the check-in period.
3. Weigh the flask and stopper and record the mass to the nearest 0.1 mg. Do not
touch the flask with your fingers after this weighing. Use tongs or wear gloves.
4. Read and record the temperature of the water.
5. Pipette 10.00 mL of the distilled water into the flask. Ask the Lab Assistant if
you are not sure about the correct technique to use pipette. Restopper the flask,
weigh it, and record the mass of the stoppered flask plus the water.
6. Add a second pipette of water to the flask. Re-stopper, weigh, and record the
weight. Repeat the entire procedure for a minimum of four readings that agree
to within a total range of less than about 0.02 g for all the readings. [If you
have what you consider 3 “good” replicates and one that seems “bad”, try the
Q-test to see if the outlying value can be rejected.] If your values seem very
 9

non-reproducible, throw out all your data and do another full set of 4 minimum,
paying closer attention to what you’re doing.
7. Correct the apparent masses of the aliquots for the buoyancy effect of
atmospheric air to get the true masses as described above. Then calculate the
true volume of the pipette using the density of water at the temperature(s) of
each aliquot.
8. Report the average “true volume” of your pipette and the associated standard
deviation of your values.
 10

Spread Sheet for Experiment 2

Item Mass
Erlenmeyer flask with stopper
First reading (Erlenmeyer flask + stopper + water)
Second reading
Third reading
Fourth reading

Item Temperature
Distilled water

Item Details
Mass of distilled water (mobserved)
Density of air (dair) 0.0012 g/cm3
Density of distilled water (dobject)
Density of weights (dweights) 8 g/cm3
Corrected mass (mcorrected)
(Show calculation)

THE DENSITY OF WATER IN g/cm3 NEAR ROOM TEMPERATURE

o
C .0 .1 .2 .5 .7 .8

18 0.998595 576 558 539 501 463 444 424

19 405 385 365 345 305 265 244 224

20 0.998203 183 162 141 099 056 035 013

21 0.997992 970 948 926 882 837 815 792
22 770 747 724 701 655 608 585 561
23 538 514 490 466 418 369 345 320
24 296 271 246 221 171 120 095 069

25 0.997044 018 0.996992 967 914 862 836 809

26 0.996783 756 729 703 649 594 567 540
27 512 485 457 429 373 317 289 261

28 232 204 175 147 089 031 002 0.995973

29 0.995944 914 885 855 796 736 706 676

30 0.995646 616 585 555 494 433 402 371

 11

EXPERIMENT 3
Quantitative Analysis of Dye in Water Sample

INTRODUCTION
Spectroscopy is a technique that uses the interaction of energy with a sample to
perform an analysis. The data that is obtained from spectroscopy is called a spectrum,
which is basically a plot of the intensity of energy detected versus the wavelength (or
frequency or wavenumber) of the energy. One might ask, “What information can be
obtained from a spectrum?” A spectrum can be used to obtain information about
atomic and molecular energy levels, molecular geometries, chemical bonds,
interactions of molecules, and related processes. Often, spectra are used to identify
the components of a sample (qualitative analysis). Spectra may also be used to
measure the amount of material in a sample (quantitative analysis).

Several instruments can be used to perform a spectroscopic analysis. In simplest

terms, spectroscopy requires an energy source (such as lamp that gives off certain
wavelengths of light, or a laser) and a device for measuring the change in the energy
source after it has interacted with the sample (often a spectrophotometer or
interferometer). In this experiment, we will employ a specific type of
spectrophotometer called single-beam UV-Vis spectrophotometer, which can provide
both ultraviolet (UV) and visible (vis) light. The idea is to shine visible light
(wavelength range between 400 to 800 nm) from a source, split it into component
wavelengths using a monochromator (mono means one; chromo means color), select
a narrow band (or color) of wavelength through a slit, then pass the monochromatic
light to a solution of a coloured compound, such as methylene blue dye (Figure 3).
The intensity of light from the monochromator (Io) is reduced as it passes through the
sample solution (I) due to the presence of light-absorbing species, in this case the
dye. The ratio between light intensities, I/Io, is related to a property of light called
absorbance, A.
 12

Figure 3: Schematic of a single-beam UV-Vis spectrophotometer.

The amount of light absorbed by the dye, A, can be used to determine the amount of
dye in the sample by comparing the absorbance data of the sample with those of
carefully prepared solutions of blue dye of known concentrations called standard
solutions using a mathematical relationship called Beer-Lambert’s Law,

Beer-Lambert’s Law: A = ɛbc [2]

where A is the absorbance, c is the concentration of standard or sample solutions, b

is the length at which light passes through (equal to the width of the cuvette or
sample holder) and ɛ is a constant called molar absorptivity (or molar extinction
coefficient). The value of ɛ depends on the wavelength used for analysis and the
nature of species in solution.

Equation 2 tells us that a plot of absorbance, A, on the y-axis against concentration of

standard solutions, c, on the x-axis will yield a straight line with a slope equal to (ɛb).
Such plot is called a standard curve, which is obtained by measuring the absorbance
signal from a series of standard solutions of known concentration. The standard curve
is then used to determine the concentration of dye (cunk) in the sample solutions by
locating the absorbance measurement of the unknown on the y-axis (Aunk) and
following a line until it intersects the standard curve. The corresponding value on the
x-axis is the concentration of substance in the unknown sample (Figure 4). This
process is called linear interpolation.
 13

Figure 4: Example of a standard curve.

The best-fitted (diagonal) line in the standard curve above is determined by a process
called linear regression.

OBJECTIVES:
i. to prepare standard solutions of methylene blue dye by dilution of stock
solutions.
ii. to generate a standard curve from absorbance of visible light data for
standard solutions.
iii. to apply Beer's law on the absorbance data in order to determine the
amount or concentration of blue dye in water sample.

EQUIPMENT/APPARATUS
UV-Vis Spectrophotometer
Volumetric flask 25 mL
Micropipette 1000 µL
Plastic cuvettes

REAGENTS/CHEMICALS
5.0 × 10-5 M stock solution of methylene blue dye
Water sample containing the blue dye
 14

PROCEDURE
Warm up the UV-Vis spectrophotometer and read the instructions for operation of
the instruments thoroughly before beginning the experiment.

Preparation of standard solutions of the methylene blue dye by dilution

1. You will be preparing four dilutions of the blue dye stock solution into separate
25 mL volumetric flasks by following steps (a) through (c) below.
(a) Using a 1000 µL micropipette, measure 1.00 mL of 5.0 × 10-5 M blue dye
stock solution prepared by the Lab Assistant and transfer into a 25 mL
volumetric flask. Dilute with water to the mark. Cap and shake well.
Transfer contents into a plastic tube and label as Standard 1. Calculate the
molarity of Standard 1 following equation 3. Record your result. Since you
took 1.00 mL of the stock and diluted it to a final volume of 25.00 mL, this
is called a 1:25 dilution.

5.0 × 10-5 M × 1.000 mL

Mstd-1 = [3]
25.00 mL

(b) Repeat 4(a) using the same pipette and stock solution, but this time
measuring 2.00 mL of the stock into another 25 mL volumetric flask. Dilute
to the line as before with water. This is called a 2:25 dilution. Cap and
shake well. Label as Standard 2. Calculate the molarity of Standard 2
following equation 3, but using the 2.00 mL volume of stock solution.
(c) Repeat the procedure above to prepare Standards 3 and 4 using 3.00 mL
(3:25 dilution) and 5.00 mL (5:25 or 1:5 dilution) volumes, respectively, of
stock solution using two clean 25 mL volumetric flasks. Calculate the
molarity of Standards 3 and 4 using equation 3 with the appropriate volume
of stock solution used for each standard.
 15

Preparing a standard curve from absorbance measurements of standard solutions

1. Fill the plastic cuvettes up to the line with each of the standard solutions. Use
the marks on the cuvette tray to remember which solution is in what cuvette.
Fill another cuvette with deionized water. This will serve as your reference or
“blank” solution, meaning one without the blue dye.
2. Run using UV-Vis spectrophotometer. Record your absorbance data. You will
use this data to construct a standard curve after the experiment.

Quantitative analysis of blue dye in water sample

1. Measure the absorbance of the water sample following the same procedure for
standard solutions.
2. Using your standard curve (don’t forget to include the line equation in your
graph) calculate the molar concentration of blue dye in the water sample.

Construct a standard curve using Excel

1. Label the x-axis as “Concentration of Blue Dye Standards, mol/L”
2. Label the y-axis as “Absorbance at A nm”
3. Use the graph title as “Standard Curve for Blue Dye”
4. Perform a linear regression (set the intercept to zero) and include the line
equation in your graph.
5. Using the line equation and the measured Absorbance (y), calculate the molar
concentration (x) of blue dye in each of your commercial samples.
 16

Spread Sheet for Experiment 3

Molarity of blue dye stock solution: _______________ mol L -1

Molar concentration and absorbance of various dilutions of blue dye:

Standard number Molar concentration (Mstd-x), mol L-1 Absorbance

1
2
3
4

Absorbance data for commercial drinks

Absorbance of unknown: ________________

Molar concentration of blue dye: ________________ mol L-1

 17

EXPERIMENT 4
Colorimetric Determination of Phosphate

INTRODUCTION
There is often a direct relationship between the intensity of the colour of a solution
and the concentration of the coloured component (the analyte species) which it
contains. This direct relationship forms the basis of the colorimetric technique. One
might readily determine the concentration of a sample based on its colour intensity,
simply by comparing its colour with those of a series of solutions of known
concentration of the analyte species. In some cases, the colour of the solution may be
due to an inherent property of the analyte itself, for example, a KMnO4 solution has a
natural purple colour, the intensity of which can be readily measured. In many other
cases, however, the solution colour is developed by the addition of a suitable reagent
which interacts with the analyte species thereby forming a coloured complex.

The amount of electromagnetic radiation in the visible region of the spectrum

absorbed by a coloured solution is often directly proportional to the concentration of
the coloured species as defined by the Beer-Lambert Law. Intensity of coloured
solutions is normally measured with a spectrophotometer.

Colorimetric techniques are useful in the analysis of a wide range of substances. One
important application is its use in determining the phosphate content of natural and
wastewater sources. Phosphate is considered to be one the most important nutrients
in natural water. Although several other nutrients (eg. carbon, nitrogen, sulfur,
potassium, calcium and magnesium) are required to facilitate growth of plant
material, particularly algae, the phosphorus content is critical in determining the level
of algal growth that the water will support. The growth of algae in natural water will
rarely occur at phosphate concentrations below 0.05 mg/dm3. Drinking water may
have a maximum allowable phosphate content of 0.3 mg/dm3, while on average, raw
sewage contains about 30 mg/dm3.
 18

The phosphate found in natural waters mainly exists as the orthophosphate species,
PO43-, however, the polyphosphates P2O74- and P3O105- are frequently encountered.
These polyphosphate species may be hydrolysed to produce the orthophosphate,
however, the species which dominates will depend on the pH prevailing in the
particular environment.

Phosphate will readily react with ammonium molybdate in the presence of suitable
reducing agents to form a blue coloured complex, the intensity of which is directly
proportional to the concentration of phosphate in the solution. The phosphate content
of an unknown water sample can be obtained by first plotting the absorbance’s of a
series of standard solutions against the corresponding concentrations, thus giving a
calibration curve. The concentration of phosphate in the unknown sample can then be
determined from the graph.

In this exercise, a sample of natural water has been provided which has been filtered
and treated to remove all materials likely to cause interference. You are required to
determine the phosphate content of the sample in duplicate, using the
spectrophotometric technique.

OBJECTIVE:
i. to introduce students to the use of UV/Vis spectroscopy in analytical
chemistry.

EQUIPMENT/APPARATUS
UV-Vis Spectrophotometer
Volumetric flask
Pipette
Measuring cylinder

REAGENTS/CHEMICALS
KH2PO4
 19

PROCEDURE
Preparation of calibration curve
Prepare a standard stock solution of phosphorus of approximately 25 mg/L P by
dissolving 2.75 mg of KH 2 PO 4 (this should be accurately weighed) in distilled water
and diluting into 25 mL volumetric flask. Use the stock solution to prepare standards
of approximately 0.20, 0.40, 0.60, 0.80 and 1.0 mg/L P, by pipetting 0.2, 0.4, 0.6, 0.8
and 1.0 mL portions respectively to separate labelled 25 mL volumetric flasks. Place
roughly 10 mL of distilled water into a 25 mL flask as a blank solution, and then
organize all the analytical solutions for colour development.

DO NOT make up the solutions to the mark yet.

Colour development
Add distilled water to all the analytical solutions (standards and samples) so that each
flask contains roughly 10 mL of solution. Starting with standard 1, add 2.6 mL of
combined reagent (prepared by Lab Assistant) using a 25 mL measuring cylinder.
Shake thoroughly and make up to the mark with distilled water. Treat all the
solutions similarly then allow 30 minutes for colour development. Run using visible
spectrophotometer.

Record your results in the table provided then plot a graph of the absorbance vs the
corresponding concentration.

Analysis of water sample

Sample solution will be prepared by the lab instructor. One group will have ONE
sample solution only.
 20

Spread Sheet for Experiment 4

Concentration Absorbance Absorbance Absorbance Average Standard

Standard
(mg/L P) 1 2 3 absorbance deviation
Blank 0.00
1 0.20
2 0.40
3 0.60
4 0.80
5 1.00

Sample

Absorbance reading for unknown solution = _____________________________

The [P] in mg/L unit = __________________________________

 21

EXPERIMENT 5
Determination of Monoprotic & Diprotic Acid
Acid-Base Titration

INTRODUCTION
Acid-Base titration involves a neutralization reaction in which a certain amount of
acid reacts with an equivalence of base. By checking the pH throughout the titration,
the sequence of titration can be followed. This is done by plotting the titration curve,
pH of the solution versus volume of the titrant added. The titrant is usually a strong
standard acid or base solution which is used to determine the analyte of basic or
acidic properties. The equivalence point in titration is the point of stoichiometric
equivalence between the analyte and the reagent. In other words, the amount of
standard reagent is equivalent to the amount of the analyte. The end point is a point
when the reaction is completed, that is when there is a change in some properties of
the solution to be determined or the point at which the addition of titrant is
terminated.

Titration Curve
A titration curve is a plot of pH versus volume of titrant. In the regular general
chemistry laboratory, this experiment is performed by measuring pH as a function of
time. The burette is set at a constant drip rate so time can be assumed directly
proportional to the volume of base or volume of titrant.

Acid Dissociation Constant, Ka

A weak acid (HA) dissociates in water according to equation 4:
HA (aq) + H2O (l)  H3O (aq) + A- (aq) ----------------------------------------------- [4]

The acid dissociation expression is:

Ka = H3O+ A- / [HA] ---------------------------------------------------------------------- [5]

From this expression we can derive the Henderson-Hasselbach equation for a weak
acid:

pH = pKa + log (A- / [HA]) ------------------------------------------------------------- [6]

 22

The reaction between a base (such as NaOH) and a weak acid is given below:
HA (aq) + OH- (aq)  A- (aq) + H2O (l) --------------------------------------------- [7]

At the "half-way to equivalence point" exactly one half of a weak acid has reacted
with hydroxide. At this point, the amount of HA remaining is exactly equal to the
amount of conjugate ion, A–, formed. The ratio [A–] / [HA] in equation [7] are equal
to one and log [1] is equal to zero. So equation [7] becomes:
pH = pKa (at the half-way to equivalent point) --------------------------------------- [8]

Indicators
Indicators are weak organic acids (HIn) that change colour when deprotonated (In-).
A few drops of indicator added to the analyte solution before the beginning an acid-
base titration. When enough base titrant is added to the analyte solution the
equilibrium expressed in equation [4] will shift towards products.
HIn (aq)  In- (aq) + H+ (aq) --------------------------------------------------------- [9]

The result is the formation of more of the deprotonated indicator (In-) and a
corresponding colour change of the analyte solution (the endpoint). A good indicator
for a specific acid-base titration has an endpoint with a pH at or near the pH of the
equivalence point.

OBJECTIVES:
i. to determine the titration curve of monoprotic and diprotic acid.
ii. to determine the acid dissociation constants of monoprotic and diprotic
acid.

EQUIPMENT/APPARATUS REAGENTS/CHEMICALS
pH meter NaOH pellet
Burette HCl solution
Beakers 250 mL Phenolphtalein indicator
Retort stand Deionized water
Magnetic stirrer Acetic Acid
Stir bar Oxalic Acid
pH paper
 23

PROCEDURE
Preparation of 0.10 M NaOH solution
1. Weight the NaOH pellet according to the Equation 10 and 11 below in order
to obtain 0.10 M NaOH solution.

M1V1 = M2V2 (10)

mass
No. of mol = (11)
molar mass

2. Dissolve the NaOH pellet in the 100 mL deionized water.

Preparation of 0.10 M HCl Solution

1. Calculate the number of mol needed for HCl solution in 50 mL volumetric
flask by using Equation 8 and 9 above.

Titration Procedure
1. Place exactly 100 mL of deionized water into a 250 mL beaker.
2. Add 8.0 mL of 0.10 M HCl solution and 3 - 4 drops of phenolphthalein
indicator. Place the beaker on a white paper to best observe the colour
changes.
3. Please get some advice from lab instructor on how to handle pH meter wisely.
Rinse the pH electrode every time before and after use.
4. Use a utility clamp to suspend a pH electrode on a ring stand as in Figure 5.

Figure 5: Titration setup with probe of pH meter.

 24

5. Place the pH electrode in the 0.10 M HCl (Strong-monoprotic acid) solution

and adjust its position toward the side of the beaker so the stir bar does not
collide with the fragile electrode.
6. Rinse a 50 mL burette with a few mL of the 0.10 M NaOH (Strong base)
solution. Use a funnel and carefully fill in the burette.
7. Titrate the solution by adding the NaOH titrant in 0.5 - 1 mL increments. The
coloured form of the phenolphthalein will begin to stay for a while and then
disappear. At this point add the NaOH dropwise until the colour changes
remain. This is the endpoint for phenolphthalein.
8. Record the volume and pH that indicator changes colour. The guideline for
colour changes can be referred in Figure 6 below. Then rinse the pH probe
with deionized water and replace the probe tip into its vial. Plot the titrant
volume (x-axis) versus pH (y-axis).

Figure 6: Setup for titration and guideline for colour changes

9. Repeat the procedure above for the:

• 4.0 mL of 0.1 M Acetic Acid (Weak-Monoprotic Acid) – 0.1 M Sodium
Hydroxide (Strong Base)
• 4.0 mL of 0.1 M Oxalic Acid (Diprotic Acid) – 0.1 M Sodium Hydroxide
(Strong Base)
10. Determine the range of end point and equivalent point based on the graph
plotted as shown in Figure 7 below.
 25

Figure 7: Titration graph for acid-base titration.

11. Discuss the differences between each graph plotted & label the following on
the four titration curve if applicable:
• Equivalent point
• pH range of the buffering region
• pH = pKa
• pH range of phenolpthalein

Caution: Students are compulsory to wear safety goggles, lab coat and safety
shoes/shoes cover at all times in lab. Acids & Bases are caustic solutions and
should be handled in fume cupboard. If spills occur, wash affected areas
immediately and inform your Lab Instructor. Wear gloves when working with
oxalic acid, as it is toxic when absorbed through the skin.

Waste Disposal – Titrated solutions can be discarded down the drain. Excess
titrant can be disposed of down the drain.
 26

Spread Sheet for Experiment 5

Part 1: HCl + NaOH Part 2: Acetic acid + NaOH

Titrant volume Titrant volume
pH pH
added (mL) added (mL)

Part 3: C2H2O4 + NaOH

Titrant volume
pH
added (mL)

*Feel free to use another papers if this spread sheet is insufficient.

 27

EXPERIMENT 6
Determination of Chloride by the Mohr Method
Precipitation Titration

INTRODUCTION
Titration is a process by which the concentration of an unknown substance in
solution is determined by adding measured amounts of a standard solution that reacts
with the unknown. Then the concentration of the unknown can be calculated using
the stoichiometry of the reaction and the number of moles of standard solution
needed to reach the so called end point.

Precipitation titrations are based upon reactions that yield ionic compounds of limited
solubility. The most important precipitating reagent is silver nitrate. Titrimetric
methods based upon silver nitrate are sometimes termed argentometric methods.
Potassium chromate can serve as an end point indicator for the argentometric
determination of chloride, bromide and cyanide ions by reacting with silver ions to
form a brick-red silver chromate precipitate in the equivalence point region.

The Mohr method uses chromate ions as an indicator in the titration of chloride ions
with a silver nitrate standard solution. After all the chloride has been precipitated as
white silver chloride, the first excess of titrant results in the formation of a silver
chromate precipitate, which signals the end point. The reactions are:

Ag+ + Cl- AgCl(s)

2Ag+ + CrO42- Ag2CrO4 (s)

By knowing the stoichiometry and moles consumed at the end point, the amount of
chloride in an unknown sample can be determined. This report describes experiments
aimed at determining the concentration of chloride in a liquid sample.

OBJECTIVE:
i. to determine the concentration of chloride in liquid sample
 28

EQUIPMENT/APPARATUS REAGENTS/CHEMICALS
Burette NaCl
Pipette 25 mL CaCO3
Pipette bulb K2CrO4
Erlenmeyer flasks 250 mL AgNO3
Desiccator
Volumetric flask 250 mL
Graduated cylinder 100 mL

PROCEDURE
Preparation of 5% K2CrO4 (indicator)
Weight 1.0 g of K2CrO4 and dissolve in 20 mL of distilled water.

Preparation of standard AgNO3 solution

Weight 4.25 g of AgNO3 and transfer to a 250 mL volumetric flask and made up to
volume with distilled water. The resulting solution was approximately 0.1 M. This
solution will be standardized against NaCl.

Preparation of NaCl solution

Weight 0.2500 g portions of NaCl into Erlenmeyer flasks and dissolve in about 100
mL of distilled water.

Titration
Pour 25 mL of NaCl solution into conical flask. Add about 1 mL of K2CrO4 and
titrate the solution to the first permanent appearance of red Ag2Cr2O4. Repeat for
another 2 times.

Determination of Cl- in liquid sample

The unknown has been prepared in 100 mL volumetric flask. Take 5 mL of NaCl
solution and mix with 20 mL tap water. Introduce about 1 mL of K2CrO4 and titrate
the solution to the first permanent appearance of red Ag2Cr2O4. Record the volume
used and determine the % w/w of Cl- in unknown sample.
 29

Spread Sheet for Experiment 6

Standardization of AgNO3

Replicate Sample, g NaCl Volume of AgNO3 Concentration of

used, mL AgNO3, M
Blank -
1
2
3

Molarity of AgNO3: _____________ M

Determination of Chloride in Unknown

Volume of AgNO3 (mL) = _____________________________

%(w/w) Cl in unknown = ______________________________

 30

EXPERIMENT 7
Determination of Amino Acids
Potentiometric Titration

INTRODUCTION
This experiment will demonstrate the acidity of the carboxyl group (COOH), the
alkalinity of the amino group (-NH2) and the acid-base characteristics of the R-group
(if any) on the amino acid. Amino acids under physiological conditions undergo
double ionization and are called "zwitterions" (twin ions). This term comes about
from the deprotonation of the carboxyl group to the carboxylate ion (COO-) and the
protonation of the amino group to the ammonium ion (-NH3+). R-groups, such as the
imidazole ring, carboxyl group, amino group, guanidino group, will protonate or
deprotonate depending upon their chemical characteristics and the pH of the solution
in which they are solvated.

OBJECTIVE
i. to determine the pKa of the unknown amino acid via potentiometry
method.

EQUIPMENT/APPARATUS
Analytical balance
Burette
Erlenmeyer flask 125 mL
pH meter

REAGENTS/CHEMICALS
HCl
NaOH
Buffer solutions of pH 4 and 7
Histidine
 31

PROCEDURE
1. Weigh 2 samples of histidine such that they are not more than 0.1000 g. Label
them as Sample 1 and Sample 2.
2. Pour each sample into the Erlenmeyer flasks the amino acid in about 25 mL
of water.
3. Check the pH of each amino acid solution.
4. Then, add enough 1 M HCl to each of the solution to adjust the pH to a pH of
about 1 to 1.5.
5. Fill the burette with 0.1 N NaOH.
6. Titrate each amino acid sample against the NaOH solution.
7. Record the pH readings at the volumes indicated in the spread sheet of this
experiment. The readings should be taken after the addition of every 0.5 mL
of 0.1 N NaOH.
 32

Spread Sheet for Experiment 7

Sample 1 Sample 2
Mass (g)

Sample 1 after _______ mL HCl added Sample 2 after _______ mL HCl added

pH

Vol. 0.1 N Vol. O.1 N Vol. 0.1 N

pH pH pH
NaOH (mL) NaOH (mL) NaOH (mL)
0.0 15.0 30.0
0.5 15.5 30.5
1.0 16.0 31.0
1.5 16.5 31.5
2.0 17.0 32.0
2.5 17.5 32.5
3.0 18.0 33.0
3.5 18.5 33.5
4.0 19.0 34.0
4.5 19.5 34.5
5.0 20.0 35.0
5.5 20.5 35.5
6.0 21.0 36.0
6.5 21.5 36.5
7.0 22.0 37.0
7.5 22.5 37.5
8.0 23.0 38.0
8.5 23.5 38.5
9.0 24.0 39.0
9.5 24.5 39.5
10.0 25.0 40.0
10.5 25.5 40.5
11.0 26.0 41.0
11.5 26.5 41.5
12.0 27.0 42.0
12.5 27.5 42.5
13.0 28.0 43.0
13.5 28.5 43.5
14.0 29.0 44.0
14.5 29.5 44.5

Sample 1 pKCOOH Sample 2 pKCOOH

 33

Sample 1 pKNH2 Sample 1 pKNH2

Average Experimentally
KNOWN Values
Determined
pKCOOH
pKNH2
 34

EXPERIMENT 8
Determination of Ascorbic Acid in Vitamin C
Redox Titration

INTRODUCTION
Redox titration or oxidation-reduction titration is a reaction between an oxidizing
agent and a reducing agent or in other words the reaction in which the reacting
species undergo a change in the oxidation number.

OBJECTIVE
i. to determine the % yield of ascorbic acid in unknown vitamin C.

EQUIPMENT/APPARATUS
Erlenmeyer flask
Rubber stopper
Aluminium foil
Volumetric flask 1000mL
Volumetric flask 250 mL
Volumetric pipette

REAGENTS/CHEMICALS
Potassium Iodide (KI)
Primary standard Na2S2O3
Starch indicator
Vitamin C pellet

PROCEDURE
Preparation of 0.05 N of iodine
Obtain a 250 mL Erlenmeyer flask with a rubber stopper and cover the stopper with a
piece of Al foil. In the hood, using a triple beam balance, weigh out 20 g of KI (you
need not be very accurate) and dissolve it in 50 mL of distilled water. Weigh out
approximately 6.4 g of I2 and dissolve it in the KI solution. Do not take iodine
crystals back to your lab bench because they give off a corrosive vapour! Once
 35

you have the iodine in solution, then you can take it back to your lab bench. Once
your I2 and I are dissolved, pour the solution into a 1000 mL volumetric, add - water
to bring the volume up to the 1000 mL mark, then mix the solution and store it in
your cabinet, out of direct sunlight. It may not be possible to completely dissolve the
I2. If you have some undissolved I2 simply decant your solution into a 1 L glass bottle
and dilute it to 1 L with distilled water. When doing this makes sure that no
undissolved I2 is transferred into your solution because this will dissolve over a
period of time and change the normality of your solution. Standardize this solution in
the next step, but do not throw away because this solution is needed in the second
part of the lab.

Preparation of 0.05 N of sodium thiosulfate

Accurately weigh about 2 g of Na2S2O3 (Primary Standard). This will be found in the
lab desiccator. Place this sample in a 250 mL volumetric and fill to the mark. Note:
This solution is not stable for long periods of time. If won’t be using this solution
within the next 24 hours, see your instructor or your text for instructions for making a
stable standard solution.

Standardization of I2
Pipette out a 25 mL aliquot of your sodium thiosulfate standard using a volumetric
pipette and add about 2 mL of the starch indicator solution. Titrate this solution
against the I2 solution. The end point is indicated by the appearance of a deep blue
colour which should persist for at least 1 min. Unlike the earlier titrations which you
have done, the endpoint in this reaction does not come instantly. Instead, as you near
the end point you will see the appearance of a blue colour which disappears when
swirling the solution. So when you reach this point in your titration add the I2
solution one drop at a time and swirl your flask vigorously. Do these until you reach
the endpoint. The dropwise addition is very important if you want to get accurate
results. When you have standardized your Iodine solution you may dispose of the
thiosulfate solution.

Determination of Vitamin C
Vitamin C spontaneously oxidizes when exposed to air. If you don't keep Vitamin C
tablets tightly closed, they will slowly lose their potency. You can see that if you ever
 36

wanted to determine the potency of Vitamin C that has sat on the shelf for a while,
this would be a reasonable procedure.

Determination of an unknown
You will be given an unknown containing approximately 1 gm of Vitamin C.
Quantitatively transfer this to your 250 mL volumetric (Make sure is was well
cleaned and rinsed since it just had Thiosulfate in it) and dissolve the Vitamin C in
250 mL of water. Since this will slowly air oxidize, keep this volumetric capped and
place it in a beaker of ice to slow the oxidation rate down. Do all titrations of your
unknown on the same day and DO NOT store the solution overnight. To titrate this
solution simply remove a 25 mL aliquot with a volumetric pipette and transfer the
aliquot to a 250 mL Erlenmeyer and add 2 mL drops of starch indicator. Titrate
directly with your Iodine solution. Do at least three titrations.

Caution:
I2 sublimes (goes directly into a vapour form) at room temperature. This vapour form
is caustic and a potential health threat. That is why you must weigh it in the hood,
and add the KI and put it into solution as quickly as possible. DO NOT HAVE
SOLID I2 SITTING ON YOUR LAB BENCH FOR ANY LENGTH OF TIME. It is
also a good idea to dispose of the Iodine solution in a sink in the hood, rather than the
sinks in open part of the lab.

Notes on Calculations:
1. Notice that the concentration of the stock solution is in NORMALITY.
Normality = Molarity * n, where n is a coefficient that takes care of reaction
stoichiometry’s. In redox reaction, n = number of electrons in the reagent’s ½
reaction. The nice thing is that N*V=N*V. No matter what the reaction
stoichiometry is. So if we had 50 ml of 0.1007 N stock solution and added to it
50 ml of H2O, our final normality is 0.05035 N. If 25 ml of this is titrated with
26 ml of our Iodine solution we then have 25 (0.05035) = 26 (X), and X =
0.05144 N.
 37

2. What is the Normality of our thiosulfate solution? In making this solution we

have 2 g of Na2S2O3 (FW=158.09) in 250 mL of water so the Molarity is
2/158.09/0.25 = 0.05061M.
The thiosulfate ½ reactions are:

2S2O32-  S4O62- + 2e-

In this equation we see that each mole of thiosulfate makes 1 mole of electrons
so N = M and N = 0.05061 N.

3. Let's assume you have a 1.0-gram sample that was 95% Vitamin C. 1.0 grams
X 0.95 = 0.95 grams. The equivalent weight of Vitamin C is 88.07 in this
reaction so 0.95 grams represents 0.95/88.07 or 10.787 mequivalents of vitamin
C. You take a 25 ml aliquot from a 250 ml volumetric, so you are actually
titrating 1.079 meq. This will require 1.079 meq of I2 solution to titrate it, so
the equivalence point should be at 1.079 x 10-3 / 0.05144 or 20.98 ml.

For the discussion, please include the % yield of ascorbic acid in vitamin C &
show the calculation step clearly.
 38

Spread Sheet for Experiment 8

Standardization of Iodine titrant:

Volume of sodium thiosulfate, mL

Concentration of sodium thiosulfate, N
Volume of iodine titrant, mL
Concentration of iodine titrant, N

Titration of unknown:
Mass of vitamin C sample: _____________ g

1st 2nd 3rd

Volume of vitamin
C solution, mL
Volume of iodine
titrant, mL
Normality, vitamin
C
Molarity, vitamin C
Moles, vitamin C
mg, vitamin C

Average mg of vitamin C in sample: ___________________ mg

Vitamin C in sample: ___________________ mg/g sample
 39

APPENDIX A

Title Jotter Lab Report

Experiment 1

Experiment 2

Experiment 3

Experiment 4

Experiment 5

Experiment 6

Experiment 7

Experiment 8
 40

APPENDIX B

EXAMPLE OF ABSTRACT!

Study was done to investigate the effect of pH on ageing

properties of natural rubber latex. Extractable protein OBJECTIVE
content and tensile properties of natural rubber latex
films were observed. Natural rubber latex was treated
EXPERIMENT
with 7.0% hydrochloric acid to get pH values in the range
of 2.0-9.0 while, 10.0% of potassium hydroxide was used
to increase the pH from 11.5 to 13.0 at 70oC. Results
showed that the extractable protein content of the films
were low at extreme acidic and alkaline conditions. While, RESULTS
extractable protein content increased after ageing
process. Generally, tensile properties i.e. tensile strength
and elongation at break were reduced after ageing
CONCLUSIO
process. Results also represents, not much different in
M100 and M300 for aged and unaged films.