Quantitative Analysis: LAB MANUAL (Phar205)

Lebanese International University
School of Pharmacy
QUANTITATIVE ANALYSIS
LAB MANUAL (Phar205)
Absorbance versus volume of Ligand
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Absorbance at 500 nm
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Absorbance at
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500 nm Collected by
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0.0 Dr. Mohammad Ahmad Assi

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volume of Nitros R (mL )

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-1- LABORATORY ITEMS
Name Description Picture
Used to hold and heat liquids.

Beaker Multipurpose and essential in the lab.
The Erlenmeyer Flask is used to heat

and store liquids. The advantage of the
Erlenmeyer Erlenmeyer Flask is that the bottom is
Flask wider than the top, so it will heat quicker
because of the greater surface area
exposed to the heat.
The Florence Flask is used for heating
substances that need to be heated
Florence evenly. The bulbed bottom allows the
Flask heat to distribute through the liquid
more evenly. The Florence Flask is
mostly used in distillation experiments.
The Volumetric flask is used to measure

Volumetric one specific volume. They are mostly
Flask used in mixing solutions and preparing
accurate concentrations.
A graduated cylinder is used to

accurately measure volumes of objects.
Graduated
Graduated cylinders are generally more
Cylinders accurate and precise for this purpose
than flasks and beakers.
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The dropper is used for moving small

Graduated amounts of liquid from place to place.
Dropper They are usually made of plastic and
are disposable
Graduated pipettes, also called Mohr

pipettes, use a series of marked lines
(as on a graduated cylinder) to indicate
different calibrated volumes. These also
come in a variety of sizes, and are used
Graduated
much like a burette, in that the volume
Pipette is found by calculating the difference of
the liquid level before and after liquid is
dispensed. Typically the precision of a
graduated pipette is not as great as that
of a volumetric pipette.
Volumetric pipettes allow the user to
measure a volume of solution extremely
accurately and then add it to something
else. They are commonly used to make
Volumetric laboratory solutions from a base stock
Pipitte as well as prepare solutions for titration.
They are typically marked to indicate
one single volume in a particular size
pipette (as are volumetric flasks). Many
different sizes are available.
The buret is used in titrations to

measure precisely how much liquid is
Buret used.
The Micro-spatula, commonly called a

Micro-spatula spatula, is used for moving small
amounts of solid from place to place.
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The stir rods are used to stir things.

Stir Rod They are usually made of glass. Stir
Rods are very useful in the lab setting.
The funnel can be used to target liquids

Funnel into any container so they will not be
lost or spilled.
The Mortar and Pestle are used to

Mortar and crush solids into powders for
Pestle experiments, usually to better dissolve
the solids.
Bottles can be used for storage, for

mixing and for displaying.
Bottle Some are wide mouthed bottles, others
are not.
A wash bottle is a squeeze bottle with a

nozzle, usually used to rinse various
pieces of laboratory glassware, such as
test tubes and round bottom flasks.
Usually, wash bottles are filled with
Washing deionized water and may also be filled
Bottle with detergent solutions and rinse
solvents such as acetone and ethanol.
When pressure is applied to the bottle,
the water inside becomes pressurized
and as a result it is forced out the
nozzle into a narrow stream of liquid.
A low density polyethylene (LDPE)

bottle supplied with a removable bellow-
type dropping pipette. The pipettes are
Dropping
graduated for nominal indication of
Bottle amount of liquid being drawn. Bellows-
shaped bulb makes for easy use and
provides excellent drop information.
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A cuvette is a small tube of circular or

square cross section, sealed at one
end, made of plastic, glass, or fused
quartz designed to hold samples for
Cuvette spectroscopic experiments.
The best cuvettes are as clear as
possible, without impurities that might
affect a spectroscopic reading.
Crucibles are used to heat small

Crucible quantities to very high temperatures.
Evaporating The Evaporating Dish is used to heat

Dish and evaporate liquids.
The watch glass is used to hold solids
Watch Glass when being weighed or transported.
They should never be heated.
Hot plates are often used in laboratory
settings to heat glassware. Some
Hot Plate hotplates also contain a magnetic
stirrer, allowing the heated liquid to be
stirred simultaneously
Bunsen burners are used for heating
and exposing items to flame. They have
Bunsen many more uses than a hot plate, but
Burner do not replace a hot plate. Wire gauze
are used on stands to hold items.
The triangle is used to hold crucibles

Triangle when they are being heated. They
usually sit on a ring stand.
Tongs are used to hold many different

Tongs things such as flasks, crucibles, and
evaporating dishes when they are hot.
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The clamp stand is used to hold

Stand equipment while they are being used.
Buret Clamp A buret clamp is used to fasten

and Holder glassware into place on a ring stand.
Ring Support A support ring

is used to hold big objects.
Test tubes are used to hold, mix, or

heat small quantities of solid or liquid
chemicals, especially for qualitative
experiments and assays. Their round
Test tube bottom and straight sides minimize
Rack mass loss when pouring, and make
them easier to clean.
The test tube rack is used to hold test
tubes while reactions happen in them or
while they are not needed.
Test tube The holder is used to hold test tubes

Holder when they are hot and untouchable.
The thermometer is used to take

temperature of solids, liquids, and
Thermometer gases. They are usually in oC, but can
also be in oF.
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Stoppers come in many different sizes.

The sizes are from 0 to 8. Stoppers can
Stopper have holes for thermometers and for
other probes that may be used.
Parafilm can be used to seal cuvettes,

Parafilm test tubes, and bottles.
Paper Towels are essential to the lab

Paper Towels environment. They will be used in
almost every lab.
Test tube The test tube brush is used to easily

Brush clean the inside of a test tube.
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-2- SAFETY REGULATIONS & Policy
Safety in the chemistry rooms is the top priority for you, the students, for your
parents, and for us. This list of rules has been developed to ensure your safety. You
are asked to follow and abide by these rules at all times.
1. Conduct yourself in a responsible manner at all times in the lab.

2. Follow all written or verbal instructions carefully.
3. Dont eat, drink, or chew anything in the lab.
4. Dont store beverages and food in the refrigerators of the lab.
5. Dont stiff or taste chemicals.
6. Never do anything in the lab not called for in the laboratory procedures or by
instructor. Do only authorized experiments.
7. Be prepared for your work in the lab. Read all the procedure thoroughly
before entering the lab.
8. Practical jokes and pranks are dangerous and prohibited.
9. Water play with distilled water bottles is never permitted.
10. Observe good housekeeping practices. Bring only your laboratory manual,
notebook, and reports. Books and bags should be left at the desk areas.
11.Read all labels well before using any chemical.
12. Use the fume hood when working with volatile substances or poisonous
vapors. Never place your head into the fume hood.
13. Dispose of all chemical waste properly. Follow specific instructions for
chemical disposal. Solid chemicals and papers are to be disposed of in the
proper waste containers, not in the sink.
14. Examine all glassware before use. Do not use cracked glassware or
damaged electrical equipments. Place broken glassware in the designated
glass disposal container carefully.
15. Never return unused chemicals to their original containers.
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16. Return all the equipments clean and dry at the end of the experiments.
17. You are responsible to keep the proper equipments in the trays. Dont look
for missing items in others trays.
18. Return all stoppers to stock bottles or containers immediately when you are
finished. Many chemicals absorb moisture from the air.
19. Never use mouth suction to fill a pipette.
20. Do not immerse hot glassware in cold water, it may shatter. Also dont add
water to acid.
21. It is not allowed to enter the lab without a lab coat, nor to work without an
instructor present.
22. Wear laboratory goggles any time chemicals,heat, or glassware are used. No
exceptions. Contact lenses should not be worn in the lab until you have
permission from your instructor.
23. Long hairs, dangling jewelry and loose. Clothing must be secured in the lab.
24. No sandals or open- backed shoes are allowed.
25. Be familiar with all safety and emergency facilities such as first aid kit, fire
extinguishers, telephone.
26. Wash your hands with soap after performing any experiment, and before
leaving the lab. Clean and rinse all work surfaces at the end of experiment.
27. Absent students are expected to inquire about any material discussed during
their absence.
28. Students must read and summarize the experiments in the notebook before
entering the lab.
29. You may be asked to answer a drop quiz before doing an experiment.
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30. Know what to do if there is a fire drill during session. Containers must be
closed, gas valves, fume hoods, and any electrical equipment must be turned
off.
31. If a chemical should splash in your eye or skin, immediately flush with water
from sink or eyewash station.
32. If a thermometer is broken, mercury should not be touched. Notify the
instructor immediately.
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-3- Laboratory Reports and Note Books
Students preparation for the experiment is usually performed in two weeks:

A = Dry Lab Week: This involves doing preparatory work done at home before
part B. You will have an entire week for part A.
B = Wet Lab Week: This is the actual experiment performed in the lab. You
are expected to spend an equal amount of time on both parts of an experiment!
Notebooks Content
The lab notebook must:

State what was done.
State what was observed
Be understandable to someone else
Include complete description of the experiment:
Purpose
Methods
Results
Conclusions
Include balanced chemical equations for every reaction used.
Paste hardcopies of important data.
Include calculations, titles, dates and table of contents.
Notebooks are legal documents and routinely used for patent litigation.
Laboratory notebook should be bound (not spiral).
All assignments should be answered in the notebooks.
Notebooks should be available with students as soon as they are present in
the lab.
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Reports Regulations
1. A laboratory report will be required for each experiment. All experiments

are performed in group work, every group should submit their own report.
2. Reports are usually due at the end of the current laboratory session unless it
is otherwise announced.
3. Reports should be sent only by emails within one week from the experiment
performance. Late reports, submitted no more than one days after the due
date, will result in a lowered
grade. Any further delay will
prevent the students report from
being graded.
4. The report should be complete,

clear, and concise. All appropriate
subject matter should be included.
The student should demonstrate
an adequate understanding of the principles involved in the investigation.
5. Reports should be neat and attractive in appearance.
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Reports Contents
All reports should contain the following information in the order given.
I. Title of experiment (presented in the front page with the date, names of
student and partners, and the name of instructor to whom the report is
subjected)
II. Short General Introduction
III. Purpose or Objective
IV. Equipments, tools and Chemical Reagents
V. Experimental procedure
Students need not to repeat the description of the procedure if it is the same as
indicated in the lab manual. However, they should summarize the procedure
using past tense and describing their work; also they should indicate any
modifications and write chemical equations where possible.
VI. Theory:
Students need to include the equations of reactions studied, and discuss briefly
the principle of experiment.
VII. Data
The data taken should be presented in a systematic way, preferably in tabular
form with units. Each Table should have a number, a title and an easy reference
number in the report. The uncertainty on every measured value must be
reported along with the unit.
VIII. Calculations and Results

This should include sample calculations and all results. Results should be
reported in tabular form whenever possible. All answers must be rounded to the
correct # of significant figures.
IX. Discussions
Include when applicable the following:
- A record of all observations (changes of temperature, colors, gas
evolution, precipitation, relative speed of reaction etc.).
- Comment on any serious source of error.
- Your critical comment or suggestion on any aspect of the
experiment.
X. References
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Quantitative Classical Chemical Analysis
Gravimetry Volumetric Titrations Instrumental analysis
Acid-base Precipitation Complexometric Redox
Titrations involving iodine (I2)

Permanganimetric Dichromatometric
Permanganimetric
Iodimetry
Iodometry
Iodometric titration of copper
Titration Analyte Titrant Indicator

example
Acid-base Quantification Acetic acid NaOH (sodium Phenolphthalein

of acetic acid in (CH3COOH) hydroxide)
a vinegar
Complexometric Water Calcium and EDTA Eriochrome black

Hardness magnesium T
(Calcium and (Ca 2+ , Mg 2+)
magnesium)
Precipitation Quantification Chlordie AgNO3 (silver Mohr, Volhard,
of chloride (Cl- nitrate) Fajans
) in water
Redox Quantification Hydrogen peroxide KMnO4 No indicator

of OCl- in (H2O2) (potassium
Bleach permanganate)
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Introduction
Measurments and Errors
Objectives
1. To learn how uncertainty in a measurement arises

2. To learn to indicate a measurements uncertainty by using significant figures
3. To learn to determine the number of significant figures in a calculated result
A. Uncertainty in Measurement
There is no absolute certain measurement. Different measurement apparatus

have varied precisions. Three things need to be remembered on scientific
measurement:
1. Every measurement is subjected to errors. The margin of errors is called
uncertainty.
2. Uncertainty cannot be avoided (random error); but minimized
3. Mistake should be avoided (systematic error).
30.56 0.01 kg, 57.8 0.1
A measurement always has

some degree of uncertainty (range
of error)
Different people estimate

differently. Record all certain
numbers and one estimated
number.
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What are the uncertainties on weights?

- Cheap balances: Measurements are trust worthy to the nearest gram 5 1
- Others can read as 5:3g, then 5.3 0.1 uncertainty.
- Other balances: 5:298g which is more precise. Measurements made with a
standard lab balance are trustworthy to the nearest milligram (0.001 g) while
the analytical balance to the nearest 0.1 mg, 5.2986 0.0001 uncertainties.
What are the considerations for the Lab Glassware?

Volumetric 10.00 ml pipitte:
T.D (25C 1)
10.00 0.01 (one meniscus)
Graduated 10.00 ml pipitte:

T.D (25C 1)
10.00 0.02 (Two menisci)
Volumetric flask 250.0 ml:

T.C (25C 1),
Used to prepare accurate concentrations
250.0 0.25 10.0 0.1 (one meniscus)
Graduated 10 ml cylinder:
T.D (25C 1)
10.0 0.1
Beakers cannot tell volumetric measurements. Why?
How to Read and Use a Buret?
- When reading a buret, it is important that your

line of sight be in a direction perpendicular to
the buret column.
- The burette should be vertical, avoid parallax.
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- All buret reading should be done using a

buret card.
- Dark line on card reflects off meniscus to
show where it is located.
- Upper limit of the black streak ought to be
placed just under the meniscus, so that the
bottom of the meniscus can be seen
distinctly against a narrow zone of white.
- Read volume associated with bottom of meniscus.
A 50 mL buret can be read to 0.04 ml or 0.03 ml. Since we have 2 minisci,

the uncertainties should be multiplied with 2.
A bubble in the nozzle of a buret will produce an inaccurate volume reading if

the bubble escapes during a titration. Use a sharp shaking of the burette while
draining.
The quickest way to get rid of bubbles is to fill the buret with titrant and open
the valve. Some bubbles may require light tapping to dislodge them.
Predict a conclusion concerning the relation between uncertainty and precision.
B. Significant Figures
These are numbers recorded in a measurement. All the certain numbers plus
first estimated number are significant ones. Significant figures serve only to
communicate the uncertainty in a measurement or calculation. They should be
stated when recording data or presenting a result. Significant figures are extremely
important when reporting a numerical value. The number of significant figures used
indicates the confidence (certainty) of that value:
2.33 2.3333
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Read the following text. What is your conclusion?
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Significant digits allow us to systematically express a degree of confidence in

a number.
Do the numbers 5000 and 5000. imply the same significance?
- 5000. contains four significant digits.

- 5000 is an ambiguous number. It contains either one, two, three, or four
significant digits.
Significant figures are the digits in a measurement that are known to be

precise and will not vary with repeated measurements. A measurement such as
35.004 g contains 5 significant figures because repeated measurements might give
the results shown at right. The 3, 5 and zeros are significant because they do not
vary. The 4 is significant because it only changes a little. All of the digits in the
readout of a digital balance or other digital apparatus (but not the calculator) are
significant.
Ordinarily, you read an analog scale to one tenth of the smallest graduation
present. Scale is graduated every 1ml, so reading should be to the nearest 0.1 ml.
The number of significant digits used implies a certain maximum error range.
Example:
- The number 101 has three significant figures and means a number between
100.5 and 101.5. The error range is 1 ( 0.5) or about 1% of 101.
- Three significant figures implies a maximum error range of 1%.
- Four significant figures implies a maximum error range of 0.1%.
- When using our calculators we must determine the correct answer; our
calculators are mindless drones and dont know the correct answer.
There are 2 different types of numbers:

Exact numbers are infinitely important. For example Dozen=12 item,
Minute= 60 sec
Measured numbers are determined with a measuring device so these numbers
have ERROR.
When you use your calculator your answer can only be as accurate as your
worst measurement.
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Rules for Counting Significant Figures
1. Nonzero integers always count as significant figures.
1457 4 significant figures
2. Zeros:
a.Leading zeros never count. Zeros at the beginning of a number are not
significant; they act only to locate the decimal point.
0.0025 2 significant figures
b.Captive zeros always count, (they are always significant).
c. Trailing zeros count only if the number is written with a decimal point
100 1 significant figure

100. 3 significant figures
Zeros at the end of a number and after the decimal point are significant. It is
assumed that these zeros would not be shown unless they were significant.
A. Which answers contain 3 significant figures?

1) 0.4760 2) 0.00476 3) 4760
B. All the zeros are significant in

1) 0.00307 2) 25.300 3) 2.050 x 103
Check Your Answers:

A. Which answers contain 3 significant figures?
2) 0.00476 3) 4760
B. All the zeros are significant in

2) 25.300 3) 2.050 x 103
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3. Exact numbers have unlimited significant figures (infinite number) since they
are100% sure. They are not obtained by measurement, but determined by
counting (3 Titrations, 4 people) or determined by definition
(1 in. = 2.54 cm). An exact number is obtained when you count objects or
use a defined relationship.
For instance is 1 foot = 12.000000000001 inches? No

1 ft is EXACTLY 12 inches.
Scientific notation is a convenient way to write a very small or a very large

number. Numbers are written as a product of a number between 1 and 10, times the
number 10 raised to power. These exponents are not sifnificant.
A calculated answer cannot be more precise than the measuring tool.

A calculated answer must match the least precise measurement.
Significant figures are needed for final answers from:
1) adding or subtracting
2) multiplying or dividing
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Rules for Multiplication and Division
The number of significant figures in the result is the same as in the

measurement with the smallest number of significant figures.
Round to the calculated answer until you have the same number of significant
figures as the measurement with the fewest significant figures.
Rules for Addition and Subtraction
The number of significant figures in the result is the same as in the

measurement with the smallest number of decimal places (least precise).
Combined operations
If products or quotients are to be added or subtracted, perform the
multiplication and division first, establish the correct number of significant figures
in the sub answer, perform the addition and subtraction, then round to the proper
number of significant figures.
8.52 + 4.1586 18.73 + 153.2 = 8.52 + 77.89 + 153.2 = 239.61 =239.6
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A student performed two trials of titration. The results were 15.50ml and 15.60ml.
What is the average end point? Apply the significant figures rules. (Ans: 15.55ml)
This is a Tricky Question!
Apply the significant figures rules to show how to prepare 500 ml of Compound X
(Mwt: 158.04g/mole) starting from 1.7698 g. (Ans: 0.02240M)
Is there any case in which results appear to be accurate but not price?
C- Accuracy and Precision

An accurate measurement is close to the true value. Often you don't know the
true value. In this case, the precision of the measurement gives you some idea of
how much you can trust the result, although a measurement can be precise but
inaccurate.
Precision is a measure of the agreement among a series of measurements. The
precision of a measurement is sometimes expressed (after statistical analysis of
data) in the following form: 0.89 0.02 g/ml
Precision depends upon the equipment used for a measurement and upon the
care with which it is made. The way in which a measurement is written down
implies something about the expected precision!
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Accuracy how close a measurement is to an accepted value (true value).

Systematic Errors (Determinate Errors): arise from a shift in all the data
points away from the accepted value. Systematic errors can be personal, methodic,
or instrumental.
Precision how close a set of measurements is to one another.
Random Errors (Indeterminate Errors): arise from a scattering of data

points value around some average value.
Determinate errors: have a definite direction and magnitude and have an
assignable cause, which mean that their cause can be determined. Theoretically
determinate error can be eliminated. Here are some examples.
- Sampling Errors results from non representative sample.
- Methodic Errors (incomplete reactions, side reactions, impure reagents).
- Instrumental Errors (Calibration, Burettes,) .
- Personal Errors (misuse of glassware or instrument, readings of numbers.
Indeterminate errors: arise from uncertainties in a measurement as
discussed above. Indeterminate error is also called random error, or noise.
Indeterminate error can be minimized but cannot be eliminated. These errors are
an experimental fact of life and are measured by the standard deviation. Small
standard deviation means good precision but says little about accuracy. Here are
some examples:
- Sampling - too small a sample or loss of material during transfer.
- Manipulation of the sample during analysis.
- Reading measurements - errors are associated with any measurement, e.g.
lines on a burette or noise in an electrical signal output.
Categorize each of the following errors as determinate or indeterminate, and further

categorize determinate errors as instrumental, operative or methodic.
An unknown being weighed is hygroscopic
One component of a mixture being analyzed quantitatively by GC reacts with
the column packing.
A radioactive sample being counted repeatedly without any change in conditions
yields a slightly different count at each trial.
The tip of the pipet used in the analysis is broken.
In measuring the same peak heights of a chromatogram, two technicians each
report different heights.
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Part I
Applications of Acid Base Titration
Exp1: Preparation of Primary and Secondary Standards
Exp2: Determination of the Solubility of Oxalic acid at
room temperature
Exp3: Acidity of Vinegar
Exp4: Determination of Stomach Antacid
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Exp One
Preparation of Primary and Secondary Standards
I- General Introduction:
Quantitative analysis is concerned with the determination of the quantity of a
certain substance in a sample. The substance to be determined in the sample is
called the analyte. There are three methods for quantitative analysis.
1. Volumetric or titrimetric analysis: They involve volume measuring
methods.
2. Gravimetric analysis: They involve weight measuring methods.
3. Instrumental analysis: They depend on measuring some physical
constants of the substances.
Accordingly, the process of titration falls in the field of volumetric technique.
II- Purpose:
1- To learn acid-base titration technique.
2- To learn how to prepare a primary standard.
3- To learn how to standardize a secondary standard.
III- Theory:
Titration is defined as the gradual addition of a
measurable volume of solution (the titant) to react exactly with
a certain amount of another substance in solution. Titration is
mainly used for determining concentrations of solutions with a
high precision. It involves a reaction between two species
whereby in one of them the concentration is known and in the
other it is unknown.
In titration, a solution is added by means of a burette to a second solution
until all the reactants in the second solution have been consumed, within the
precision desired. In this operation, the solution being added from the burette
called titrant. The end of titration is often recognized by the color change of an
indicator that has been added.
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What is a standard solution?

The solution of known concentration is called the standard solution. A
standard solution can be of two forms:
1- Primary standard.
2- Secondary standard.
A secondary standard solution is standardized by titration technique from a
primary standard whose concentration is exactly known from weight and volume.
The term standardized means: To determine the exact concentration.
What are the properties of a primary standard?

The primary standard solution should be prepared from substances those
posses the following properties:
a- Non a hygroscope (does not absorb moisture).
b- Stable (mainly solid) not affected by the laboratory environment (heat).
c- Highly pure and of known composition.
d- Easily dissolved in water.
e- With High molecular weights (to minimize relative errors).
Oxalic acid. benzoic acid, potassium hydrogen phthalate, sodium

carbonate, and borax are examples of primary standards.
What are the characteristics that the reacting substances and the reaction should
posses so that they can use in titration technique?
a- The reaction must go nearly to completion (irreversible), the equilibrium

between reactants and products must be highly favored for products so that
only a negligible excess of standard solution is required to obtain complete
reaction within the precision, of the substances being titrated.
b- The reaction must be stoichiometric; there must be a definite relationship
between the quantities of substances reacting.
c- The reaction must be instantaneous and fast.
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d- The most important requirement for a valid titration is that there must be a
means to recognize the completion of titration within the desired precision.
Simply, suitable indicator must be found. One type of indicators is the visible
indicator that has a pH change in color in the range of end-point of reaction.
What is the difference between end point and equivalence point?

Acid-Base titration technique involves experimentally volume measurement when
a change in color occurs. This point is called the end-point in titration. The end-point is
not the equivalent point (it is one drop more). Equivalent point is the point when all the
reactants has changed to products, while the end-point is the point when all the reactants
has changed to product and the extra drop of one reactant react with the indicator to
change color.
In potentiometric titrations (based on pH measurement), the equivalence point
is indicated by a sudden, sharp rise in the pH.
What is the Titration Error?

The difference between the theoretically calculated equivalent point and the
practically determined endpoint is called the titration error. The smaller the titration
error, the more accurate is the titration. We can minimize the error by adding a small
number of indicator drops or by carrying out a blank titration.
What is the Acid-Base Indicator?

An acid-base indicator is a weak acid whose acid and base forms have different
colors, then it can be symbolized: (HIn). The ionization of such indicator in water is
represented in the following equation.
HIn + H2O (aq) H3O + (aq) + In - (aq)

The acid form of the compound (HIn) has one color and the conjugate base (In- ) has
another color. According to Le Chateliers principle, adding acid (H +) or base (OH -)
will shift the reaction one way or the other, causing the color of the indicator to
change (depending on whether HIn or In- is the predominant species.
HIn + OH- (`- +*+-*
aq) H2 O (aq) + In- (aq)
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The best indicator is the

one whose color range
includes the equivalence pH,
at best, in its mid - range.
Because this is just like any

other weak acid, you can write
an expression for Ka for it. We
will call it Kind to stress that
we are talking about the
indicator.
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Rearranging the above equation, then taking the log of both sides, we get:
pH= pK In + log(In- )/(HIn)
To see the color of acid form, the ratio [In-]/[HIn] must be equal to or less
than 0.1.
To see the color of base form, the ratio [In-]/[HIn] must be equal to or greater
than 10.
Then, the pH transition range of indicators = pKIn 1.
For example, pK in of phenolphthalein= 9.3, then the indicator pH transition range is
8.3 10.3. (Check the previous figures)
(HIn) predominates 8.3 10.3 (In-) predominates
III- Equipments and Chemical Reagents:

All volumetric analysis is ultimately based on a weight measurement, namely the
weight of a primary standard; hence an accurate analytical balance is needed. For the
purely volumetric manipulations specialized glassware is used. Since temperature has an
effect on volume, such glassware is calibrated for use at room temperature (20o C).
1- Volumetric flask: used for making up solutions to
a given volume calibrated to contain (TC) the
volume specified.
2-Pipette: used to transfer accurately measured small
volumes of solution, calibrated to deliver (TD).
3-Burette: used for dispensing any volume, fractional
within the capacity of burette, graduated in
division, calibrated to deliver.
A volumetric flask, pipit or buret, is

read by looking horizontally at the
bottom of the meniscus of the liquid.
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IV- Procedure:
Oxalic acid H2C2O4.2H2O (Mwt=126g/mole) is a primary standard reagent,
fairly stable under ordinary conditions and can be easily brought to high purity.
1- Preparation of primary standard acid solution:

1. Calculate the mass necessary to prepare 50ml of oxalic acid solution 0.05
mol.L-1. Weigh mass m of H2C2O4.2H2O, close to the calculated value by
using the analytical balance.
Weight (H 2 C 2 O 4 )g 1
2. Transfer the acid, through a funnel, into a C M
Mm V solution
clean 50 ml volumetric flask. To ensure
quantitative transfer, dissolve in distilled water.
3. Fill the flask to about one-half with distilled water. Dissolve the powder
then complete to the mark.
4. Weigh about 1g of solid NaOH. Note that Mwt of NaOH is 40 g/mole.
5. Transfer to a 250 ml volumetric flask. Add about 150 ml distilled water
and shake until the sample is completely dissolved. Then add distilled
water to the flask until the bottom of the meniscus coincides with the etched line
on the neck of the flask. Add the last 0.5 ml with a medicine dropper. Insert a
stopper in the flask and mix the solution thoroughly.
2- Standardization of NaOH solution:

1. Clean your burette and then rinse twice with 5 ml NaOH solution.
2. Fill the burette with NaOH solution and drain enough liquid so that the entire
tip of the burette is free of air bubbles. Adjust the upper level of the solution
so that the meniscus rests on the zero mark.
3. Pipette 10.00 ml of the standard solution (H2C2O4.2H2O) into an Erlenmeyer
flask. Add about 3 drops of phenolphthalein indicator.
4. Titrate with NaOH solution prepared until the color start changing from
colorless to pink. The end point volume of this titration is V1.
5. Repeat this experiment a second time to have close volume to V1.
H2C2O4.2H2O + 2 NaOH Na2C2O4 + 4 H2O
Caution: The oxalate solution is toxic, wash your hands carefully before leaving the lab.
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Data Table
Include uncertainties on all recorded measurements
Initial burette reading ml

Preliminary titration Final burette reading ml
Volume of NaOH ml
First titration Final burette reading ml
Volume of NaOH ml
Second titration Final burette reading ml
Volume of NaOH ml
Questions
1- Determine the molar concentration of the H 2C2O4.2H2O solution.

2- Determine the molar concentration of standardized solution (NaOH).
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Exp Two
Determination of the Solubility of Oxalic acid at
room temperature
I-Introduction:
Solubility is defined as the amount of substance that passes into solution to achieve
a saturated solution at constant temperature and pressure. It is expressed in terms of
maximum volume or mass of the solute that dissolve in a given volume or mass of a
solvent. Pharmacopoeias give solubility in terms of the number of parts by volume
of solvent required to dissolve one part by weight of a solid, or one part by volume
of a liquid.
Determining the solubility of drug candidates is important in pharmaceutical
research, both for the discovery phase and the development phase.
In the pharmaceutical literature, two commonly used solubility terms are:

1) Kinetic solubility: the concentration of a compound at the time when an induced
precipitate first appears in the solution.
2) Equilibrium (or thermodynamic) solubility: the concentration of a compound in a
saturated solution when an excess of solid is present, and the solution and solid are
at equilibrium.
II- Purpose:
1- To learn acid-base titration technique.
2- To determine the solubility of compounds at specified conditions.
III- Procedure
The shake-flask method for determining equilibrium solubility of oxalic acid dihydrate
at room temperature:
1- Transfer gradually 1.6 grams of oxalic acid to 10 ml of stirred distilled water in a

50 ml Erlenmyer flask and keep stirring on a magnetic stirrer for 15 minutes.
2- Stop stirring and leave the solution without agitation for 10-15 minutes.
4- Filter the suspension; transfer 3 ml of filtrate to 50 ml volumetric flask and
complete to the mark with distilled water.
5- Titrate 10 ml with 0.1 M NaOH using phenolphthalein as indicator.
Calculate the molarity of oxalic acid, then the solubility (S) as grams per 100 ml of
solution at room temperature using the following formula:
Oxalicacid molarityxM WtxDFx100

S= g / 100ml
1000
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Exp Three
Acidity of Vinegar
I-Introduction:
Vinegar is a sour liquid consisting mainly of acetic acid
and water. Acetic acid (CH3COOH or HC2H3O2) is the source
of the acidity in vinegar. Acetic acid (ethanoic acid) is an
organic acid (carboxylic acid) and is classified as a weak acid.
Vinegar is regulated by the Food and Drug Administration

(FDA) and to be legal must have a minimum of 5% acidity.
II- Purpose:
1- To determine the amount of acidity (concentration of acetic acid) in
vinegar.
2- To determine the fraction of dissociation of acetic acid.
III- Theory:
The principle acid present in vinegar is acetic acid CH 3COOH (or HC2H3O2
abbreviated by HAc). According to standard laws and regulations it should contain
at last 4 g of acetic acid per 100 ml vinegar. The total quantity of acid can be
determined by titration with standard base using phenolphthalein indicator.
Although other acids are present, the result is calculated as acetic acid. In the
experiment you will find the % by weight of acetic acid of different vinegar
brands. Moreover, the degree of dissociation or fraction of dissociation of a weak
acid can be calculated from the pH and total acidity. The total acidity of vinegar is
easily determined by titration with the titrated NaOH solution. The activity of
vinegar is commonly expressed in terms of acetic acid, its principal acid
constituent. The % is expressed in g acetic acid in 100 ml of the solution.
CH3COOH + NaOH CH3COONa + H2O
Because the reaction stoichiometry is one mole of acetic acid reacting with
one mole of sodium hydroxide, the number of moles of acetic acid in the portion of
the vinegar sample titrated can be found from equation:
moles of acetic acid = (MNaO) (VNaOH)
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35 | P a g e
MNaOH is the molarity of the sodium hydroxide solution in moles/L and VNaOH is
the volume of sodium hydroxide solution in liters.
The grams of acetic acid in the vinegar sample can be calculated by

multiplying the moles of acetic acid by the molecular weight of acetic acid
(MW). This is shown in equation:
grams of acetic acid = (moles of acetic acid)(MW)
By assuming the density of the vinegar sample to be 1.00 g/ml, the grams
of vinegar solution can be calculated by multiplying the volume of vinegar by the
density.
grams of vinegar = (volume of vinegar)(density of vinegar)
The percent by mass of acetic acid in the vinegar sample can be found
from equation:
% acetic acid = (grams of acetic acid / grams of vinegar) x 100
IV- Procedure:
1. Obtain a light colored - sample of vinegar. Record the vinegar sample name.
2. Measure the pH of your vinegar solution.
3. Re-fill your burette with your previously prepared standard NaOH solution.
4. Pipet 10 ml of vinegar sample into a 100 ml volumetric flask and make up
to the mark with distilled water.
5. Mix well, and then pipet 10.00 ml of this diluted solution into a clean rinse
100 ml Erlenmeyer flask.
6. Add 2 drops of phenolphthalein indicator. Titrate slowly with NaOH
solution till a faint pink color persists for a few seconds and doesn't
disappear.
7. Read and record the burette reading.
8. Repeat the titration a second time on an additional aliquot.
9. Determine the acidity of vinegar as % mass acetic acid/volume of the
solution. (Molar mass of CH3COOH= 60 g.mol-1).
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Data Table
Vinegar sample Name: __________
N Volume of vinegar Initial burette Final burette Volume of NaOH
reading reading
1
2
3
Average
Questions
1- Assuming that all acid to be acetic acid, determine concentration of acetic acid.
2- Determine the fraction of acetic acid dissociation from the pH and molarity.
3- Calculate the weight in grams of acetic acid per liter of vinegar.
4- Calculate the grams of solution per liter of vinegar, using the density of vinegar
1.005 g/ml.
5- Find the percentage by weight of acetic acid in your sample.
6- What is the difference between your value and the federal standard of 4%?
7- Comment on the significance of this difference.
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Exp Four
Titration of Stomach Antacid
I- Purpose:
1-To practice back titration.
2- To determine the neutralization capacity of commercial stomach antacid
tablets.
II- General Introduction:
Antacids tablets are probably one of the most widely used self-prescribed
medicine. They are taken to relief the medically undefined conditions of heartburn
or acid indigestion and sour stomach. Although this gastric distress is often
attributed to excess production of HCl, sometimes HCl is not responsible for the
symptoms and gastric HCl production may be less than normal, and acid
indigestion may be due to overeating or an irritating food.
Accordingly antacids if taken haphazardly they become bad, since as have
been mentioned indigestion might be not due to excess gastric HCl. Moreover if
the pH of stomach rises very high, the entire digestive process may be hindered.
For example, the digestion of proteins is catalyzed by the enzyme pepsin which is
deactivated if the pH of the stomach is higher than a value of 4.Therefore, only an
amount of HCl in excess of what the body of a healthy individual normally
secretes following a meal should be neutralized by the antacid and this excess
amount is usually 10 millimol of HCl per hour greater than that of HCl
production. Therefore one dose of an antacids products should react with no
more than 10 millimol HCl per hour.
This raises the question whether people who regularly consume the dose
recommended on the labels of various antacid products are in fact interfering with
normal digestion by taking doses that neutralize much more than 10 millimol of
HCl or that are capable of raising the pH of stomach contents above pH 4.
Acid-neutralizing capacity or ANC in is a measure for the overall buffering
capacity against acidification. ANC is defined as the difference between cations
of strong bases and anions of strong acids, or dynamically as the amount of acid
needed to change the pH value from the sample's value to a chosen different
value.
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III- Theory:
Antacids tablets are basic substances, so direct titration would require

an acidic titrant. However many of the antacids tablets do not dissolve in
water and to overcome this difficulty a back-titration procedure will be
followed.
This back-.titration method involves the addition of the excess strong
acid HCl to the antacid tablet and the mixture is heated to ensure complete
reaction. Some of the HCl is consumed with the antacid tablet. The antacid
and HCl solution is then titrated with NaOH to determine how much HCl
remain unreacted. The number of moles of HCl neutralized by the antacid
(HClneutralized) is the difference between the moles of HCl initially present in the
excess (HClinitial) and the moles of HCl titrated by the NaOH (HCltitrated).
HClinitial HCltitrated = HClneutralized
This method gives an accurate measurement of the neutralization

capacity of antacid tablets. It also resembles the way the tablet reacts with
HC1 in the stomach. The indicator thymol blue (which is diprotic and has two
color transitions regions) is a good choice for most antacids. Phenolphtalein
can be also used and a reasonably sharp end-point is obtained.
Various brands of antacids tablets will be available. Analyze one brand
of tablets as your directs.
For example, a brand of antacid tablets has the active ingredient
NaAl(OH)2CO3 and reacts with HCl as such:
NaAl(OH) 2CO3(s) + 4HCl NaCl + AlCl 3 + 3H2 O + CO 2
A second brand has a mixture of two active ingredients NaHCO 3 and

Mg(OH) 2 and each reacts as such:
NaHCO3 (s) + HCl NaCl + H2O + CO2

Mg(OH) 2 (s) + 2HCl MgCl 2 + 2H2O
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IV- Procedure:
1- Obtain an antacid tablet and weigh it accurately using the analytical balance.
2- Crush the antacid tablet using a mortar and a pestle. Weigh the crushed tablet
again to the nearest precision of your balance.
3- Transfer the weighed crushed tablet to a clean 250 ml Erlenmeyer flask.
4- Add 15.00 ml of 1M HCl using a pipette.
5- Heat gently on a hotplate until all the effervescence has ceased. Boil for 1-2
minutes more. Some of the inactive tablet material may not dissolve,
however, this will not interfere with the titration.
6- Cool the solution to room temperature by immersing it in a container of tap
water. When the solution has cooled add 4 drops of phenolphthalein
indicator.
7- Titrate with the standardized NaOH solution. Permanent pink color appears
at the endpoint. Since the liquid is cloudy, the color change at the endpoint
may be particularly hard to detect. Use a white reference sheet to avoid
confusion.
Note: Why is heating necessary?

- Heat removes CO2 made in HCl/antacid reaction
CaCO3 + 2 HCl CaCl2(aq) + H2O + CO2
- Removing CO2 allows maximum amount of acid to be neutralized.
Note:
- Amount and concentration of Acid could be changed based on the
antacid content.
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Data Table
Active ingredient
mg of active ingredient (from the manufacturer Label)
weight of the tablet
weight of one half of the tablet
Molarity of HCl
Volume of HCl added to tablet
Molarity of NaOH
Initial buret reading
Final buret reading
Questions
1- Write the chemical equation for the reaction of HCl with active
ingredient in the tablet.
2- Calculate the millimol of HCl solution added to tablet.
3- Calculate the millimol of NaOH solution that neutralized the excess
acid.
4- Calculate the millimol of HCl that was neutralized by the antacid
tablet.
5- Calculate the millimol of active ingredient.
6- Determine the actual value of the neutralization capacity of the antacid.
7- Find the theoretical value of the neutralization capacity of this
antacid, and compare it to the actual value.
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Experiment Five
pH Measurements and Buffer Capacity
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Experiment Five
pH Measurements and Buffer Capacity
The buffer capacity refers to the maximum amount of either strong acid or
strong base that can be added before a significant change in the pH will occur.
This is simply a matter of stoichiometry.
The ability of a buffer system to resist pH changes is its buffer capacity
and indicated by the buffer index ():
= A/pH
where: A = Molarity change of strong acid or base
= change in pH
The greater the buffer capacity, the smaller is the change in pH from
addition of a given amount of strong acid or base, the more efficient is the buffer.
Buffer capacity is dependent on the total concentration of the buffer system and
on the HA/A- ratio.
The buffer index number is generally experimentally derived in a manner

like a titration. For example, when 0.03 moles of NaOH was added to an acetate
buffer system prepared at 0.1 M, the pH expectedly increased from 4.76 to 5.03; a
change of 0.27 pH units. Therefore, the equation = B/pH = 0.03-0.00/0.27 =
1/9 = 0.11.
Buffer Capacity Table
Table
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Buffer capacity is maximum when the two components of the buffer are
present in equal amounts. The ratio of base to acid will be equal to one. Then:
pH=pKa. At such ratio the resistance of the buffer towards the addition of stong
acid or base is equal.
Buffer capacity is maximal when pH = pKa and is acceptable in the range pH = pKa 1
Buffer capacity is increased by the following factors:
- increasing the concentration of the buffer system components (e.g. doubling

the total molar concentration of the buffer system will double the buffer
capacity at a given pH).
- using equi-molar concentrations of the acid (HA) and its conjugate base (A).
Buffer capacity is maximal when pH = pKa ([HA] = [A-]).
Why use buffers in pharmacy?
- Solubility: The ionized form of a drug is more water soluble than the
unionized form. Buffers can be used to maintain a drug in its ionized (salt)
form for aqueous solutions.
- Absorption: The unionized form of a drug is more lipid soluble than the
ionized form. The unionized form therefore penetrates biological membranes
much more efficiently than the ionized form. Buffers can also be used to
maintain the drug in its unionized form.
- Stability: pH can affect the stability of a drug in an aqueous solution. For
example, ester drugs are very susceptible to hydrolytic reactions. Buffering
formulations at low pH (pH 3-5) can reduce the rate of hydrolysis.
- Tissue irritation: High or low pH can cause tissue irritation. Buffering a

formulation to near neutral pH can reduce tissue irritation. Ophthalmic
products are least irritating at pH 7-9.
Finally, Buffers are prepared in pharmacies or by drug manufacturers. They

adjust the pH of aqueous solutions for applications that require predictable
stability and best clinical outcomes. From a pharmacological perspective, it is
important to control the pH of a solution to minimize drug degradation, to
improve patient comfort and compliance, and to improve the efficacy of
delivery.
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II- Purpose:
1- To review the concepts of pH measurements.
2- To learn the use of a pH meter.
2- To measure the pH of different solutions and compare the
measurements with the theoretical prediction.
III- Theory:
Let us consider different types of solutions and go over the calculation of the
hydrogen ion concentration [H +] (and subsequently the pH) for each of them.
1- Strong Acid: pH of a 0.100 M HCl solution:

HCl H+ + Cl-
Initially 0.100 - -
After complete dissociation - 0.100 0.100
Thus [H +] = 0.100 M pH= -log[H+] = 1.00
2- Strong Base: pH of a 0.100 M NaOH solution:

NaOH Na+ + OH-
Initially 0.100 - -
Thus [OH - ] = 0.100 M pOH= -log[OH - ] = 1.00
We know that: pH + pOH = 14.00 pH = 14.00 1.00 = 13.00
3- Weak Acid: pH of a 0.100 M CH 3COOH (HAc) solution:

HAc H+ + AC-
Initially 0.100 - -
Change -x +x +x
At equilibrium 0.100 x +x +x
For HAc, the acid dissociation constant Ka is:
[H ][Ac1 ]
Ka ; K a 1.75 105
[Hac]
[HAc]
haC]
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5 x2
1.75 10
0.100 x
x << 0.100 (Ka very small) it is negligible with respect to 0.100

1.75 10-6 = x x= 1.32 10-3
Thus [H+] = 1.32 10-3 M pH= -log[H+]= 2.88
4- Weak Base: pH of a 0.100 M of NH3 solution:

NH3 + H2O NH4+ + OH-
Initially 0.100 - -
Change -x +x +x
At equilibrium 0.100 x +x +x
For NH3, the acid dissociation constant Kb is:

[ NH4 ][OH ]
Kb ; K b 1.75 105
[ NH3 ]
x2
1.75 105
0.100 x
x << 0.100 it is negligible with respect to 0.100

1.75 10-6 = x x= 1.32 10-3
Thus [OH-] = 1.32 10-3 M pOH= -log[OH-]= 2.88
5- Salt of a strong Acid and a Strong Base: pH of a 0.100 M NaCl solution

NaCl Na+ + Cl-
Initially 0.100 - -
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We can see that NaCl is not a source of H+ or OH-, thus [H+] is obtained
solely from the dissociation of water:
H2O H+ + OH-
Initially - -
Equilibrium +x +x
The water dissociation constant Kw is:

Kw = [H+][OH]; Kw = 1.00 10-14
1.00 10-14 = x2 x = 1.00 10-7
Thus [H+] = [OH-] = 1.00 10-7 M pH = -log[H+] = 7.00
6- Salt of a Weak Acid: pH of a 0.100 M NaAc solution:

NaAc Na+ + Ac-
Initially 0.100 - -
Now, we should consider the hydrolysis of Ac-:
Ac- + H2O HAc + OH-
Initially 0.100 - -
Change -x +x +x
At equilibrium 0.100-x +x +x
The base dissociation constant for the acetate ion, Kb is:

[HAc][OH ] K
Kb
; K b w 5.711010
[Ac ] Ka
x2
5.711010
0.100 x
x <<0.100 it is negligible with respect to 0.100

5.71 10-11 = x2 = 7.56 10-6
Thus [OH-] = 7.56 10-6 M pOH = -log10[OH-] = 5.12
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7- Salt of a Weak Base: pH of a 0.100 M NH 4Cl solution:

NH4Cl NH4+ + Cl-
Initially 0.100 - -
Now, we should consider the hydrolysis of NH4+

NH4+ + H2O H3O+ + NH3
Simplified as:
NH4+ H+ + NH3
Initially 0.100 - -
Change -x +x +x
At equilibrium 0.100-x +x +x
The acid dissociation constant for the ammonium ion Ka is:
[H ][ NH3 ] K
Ka
; K a w 5.711010
[ NH4 ] Kb
10 x2
5.7110
0.100 x
x <<0.100 it is negligible with respect to 0.100

5.71 10-11 = x2 x= 7.56106
Thus [H+] = 7.56 10-6 M pH = -log[H+] = 5.12
8- Salt of a Weak Acid and a Weak Base: pH of a 0.100 M NH 4Ac solution:

NH4Ac NH4+ + Ac-
Initially 0.100 - -
Here we have a source of H+ from NH4+ and a source of OH- from Ac-
1
It can be shown that, is this case, pH (pKa1 pKa 2 ) , where, pKa1 is that
2
for HAc = 4.76, pKa2 is that for NH4+ = 9.24, then:
4.76 9.24
pH 7.00
2
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9- Buffer solution:
A buffer solution is one that can resist small additions of strong acid or base
without appreciably changing its pH. Buffer solutions are used to maintain the pH of
specific chemical reactions at a nearly fixed value, such as notably in biochemical
metabolitic reactions.The composition of a buffer solution involves the presence of a
weak acid and the salt of its conjugate base (such as a mixture of HAc and NaAc) or
a weak base and the salt of its conjugate acid (such as a mixture of NH3 and NH4Cl).
Think about it!

How do buffer solutions work?
What is the effect of dilution on pH of buffers?

- Small dilution would not affect the pH of the buffer.
- Excessive dilution would change the pH of the buffer.
Why? This is because water is amphoteric.
What is the effect of temperature on pH of buffers?
- Increasing the temperature would not change the pH of the acidic buffers.
- But the pH of basic buffers is affected.
Why?
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How do we prepare buffer solutions?

In preparation of a buffer solution of certain pH, the following steps are
followed:
i. Choose an acid whose pKa is as close as possible to the required pH of the
buffer.
ii. Calculate the ratio of base to acid from the buffer equation.
10- How do we calculate the pH of buffer solutions?
a- Equimolar solution: 50.00 ml of 0.500 M HAc + 50.00 ml of 0.500 M NaAC:

50.00 0.500
Initial molarity of HAc in the solution is: M HAc 0.250M
100.00
50.00 0.500
Initial molarity of NaAc in the solution is: M NaAc 0.250M
100.00
[ NaAc]
From the Henderson-Hasselblch equation: pH pKa log10
[HAc]
pH= 4.76 + log 1.00 pH= 4.76
b- 70.00 ml of 0.0500 M HAc + 30.00 ml of 0.500 M NaAc

70.00 0.500
Molarity of HAc in the solution is: M HAc 0.350M
100.00
30.00 0.500
Molarity of NaAc in the solution is: M NaAc 0.150M
100.00
[ NaAc]
[HAc]
pH= 4.76 + log10 0.428 pH= 4.39
c- 30.00 ml of 0.0500 M HAc + 70.00 ml of 0.500 M NaAc

30.00 0.500
Molarity of HAc in the solution is: M HAc 0.150M
100.00
70.00 0.500
Molarity of NaAc in the solution is: M NaAc 0.350M
100.00
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[ NaAc]
[HAc]
pH= 4.76 + log10 2.33 pH= 5.12
Calculation of the pH of a Buffer Solution after Addition of a Small Amount of

Acid:
When a strong acid (H3O+) is added to a buffer solution, the conjugate base
present in the buffer consumes the hydronium ion converting it into water and the
weak acid of the conjugate base:
A- (aq) + H3O+ (aq) --> H2O + HA(aq)
This results in a decrease in the amount of conjugate base present and an

increase in the amount of the weak acid. The pH of the buffer solution decreases by
a very small amount because of this ( a lot less than if the buffer system was not
present).
Example: If 50.0 ml of 0.100 M HCl was added to a buffer consisting of 0.025

moles of sodium acetate and 0.030 moles of acetic acid. What is the pH of the
buffer after the addition of the acid? Ka of acetic acid is 1.7 x 10-5.
First, write the equation for the ionization of acetic acid in water and the
related Ka expression rearranged to solve for the hydronium ion
concentration.
CH3COOH(aq) + H2O(l) --> H3O+(aq) + CH3COO-(aq)
[H3O+] = Ka [CH3COOH]
[CH3COO-]
Second, make an "ICE" chart. Let "x" represent the hydronium ion
concentration once equilibrium has been re-established. We will assume that
all of the added acid is consumed.
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CH3COOH(aq) H3O+(aq) CH3COO-(aq)

Initial Amount 0.030 moles (0.0500 L)(0.100 M) = 0.0050 moles 0.025 moles
Change in Amount + 0.005 moles -0.005 moles - 0.005 moles
Equilibrium Amount 0.035 moles x 0.020 moles
Substitute into the Ka expression and solve for the hydronium ion
concentration. Convert the answer into pH.
[H3O+] = (1.7 x 10-5)(0.035/0.020) = 2.975 x 10-5

pH = 4.53
IV- Procedure:
A- Chemicals and Setup:
Reagents: Standard Buffer solutions (pH = 4.00, 7.00, 10.00), 0.100 M HCl,
0.100 M NaOH, 0,100 M HAc, 0.100 M NH3, 0.100 M NaCl, 0.100 M NaAc, 0.100
M NH4Cl, 0.100 M NH4Ac, 0.500 M HAc, 0.100 M NaAc, 1.00 M HCl, and 1.00 M
NaOH.
Glassware: Two 50.00 ml burets, 1.00 ml graduated pipet, 50 ml beaker,
three 100 ml beakers.
B- Calibration of the pH meter

Learn, with the help of your lab instructor, the principles and procedure for the
calibration of the pH meter instrument.
C- pH measurements
1- Measure the pH of a 0.050 M HCl solution.
2- Measure the pH of a 0.050 M NaOH solution.
3- Measure the pH of a 0.050 M HAc solution.
4- Measure the pH of a 0.050 M NH3 soluiton.
5- Measure the pH of a 0.050 M NaCl solution.
6- Measure the pH of a 0.050 M NaAc solution.
7- Measure the pH of a 0.050 M NH4Cl solution.
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8- Buffer solutions:
i. Work in pairs throughout the experiment. Each pair will perform only
one experiment on one buffer solution. But the results should be
recorded from other groups.
ii. Obtain two burets. Collect in two 100 ml clean and dry beakers the
necessary volume of 0.500 M HAc and 0.500 M NaAc solutions.
iii. Label the burets and fill one of them with the HAc solution, and the
other with the NaAc solution.
* Solution A :Equimolar (50-50) mixture

a- Empty from the two burets 25.00 ml of HAc and 25.00 ml of NaAc
respectively, into a clean 50 ml, beaker. Mix well.
b- Pour 25.0 ml of this buffer solution into a 50 ml beaker. Measure the pH.
c- Add to this solution 0.25 ml, of 1.00 M HCl and read its pH after good
stirring.
d- Add another 0.25 ml, of HCl, stir well and read the pH.
e- For the remaining buffer, measure the pH.
f- Add to this solution 0.25 ml, of 1.00 M NaOH and read its pH after good
stirring.
g- Add another 0.25 ml, of NaOH, stir well and read the pH.
* Solution B: 35-15 mixture
Repeat steps a through g but using 35.00 ml of HAc and 15.00 ml of
NaAc in step a.
* Solution C: 15-35 mixture
Repeat steps a through g but using 15.00 ml of HAc and 35.00 ml of
NaAc in step a.
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Data Table
A- PH Measurements:
Solution Measured pH Theoretical pH % Error
0.050 M HCl
0.050 m NaOH
0.050 M HAc
0.050 M NH3
0.050 M NaCl
0.050 M NaAc
0.050 M NH4Cl
B- pH of a Buffer solution and its variation upon addition of strong acid or

strong base:
Solution A Measured pH Theoratical pH
Solution A (equimolar)
Solution A + 0.25 ml of 1.00 M HCl
Solution A + 0.5 ml of 1.00 M HCl
Solution A + 0.25ml of 1.00M NaOH
Solution A + 0.5 ml of 1.00 M NaOH
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Solution B Measured pH Theoratical pH

Solution B (35-15)
Solution B + 0.25 ml of 1.00 M HCl
Solution B + 0.5 ml of 1.00 M HCl
Solution B + 0.25ml of 1.00M NaOH
Solution B + 0.5 ml of 1.00 M NaOH
Solution C Measured pH Theoratical pH

Solution C (15-35)
Solution C + 0.25 ml of 1.00 M HCl
Solution C + 0.5 ml of 1.00 M HCl
Solution C + 0.25ml of 1.00M NaOH
Solution C + 0.5 ml of 1.00 M NaOH
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Questions
1- Comment on the variation of the pH of each of the above buffer solutions, upon
addition of acid or base. Which is the strongest buffer?
Plot the Buffer capacity versus pH of the prepared buffers.
2- A student is asked to prepare a buffer solution at pH = 8.60, using one of the

following weak acids: HA (Ka = 2.7 10-3),HB (Ka = 4.4 10-6),
HC (Ka = 2.6 10-9). Which acid should she choose? Why?
3-A buffer solution is prepared by adding 50.0 ml of 0.220 M NaOH to 100.0 ml of

0.15 M acetic acid, HAc. What is the pH of the resulting buffer solution?
4- What is the pH of the buffer formed by mixing 100.0 ml of NaH2AsO4 0.1 M

with 50.0 ml of NaOH 0.12 M.
5- How would you prepare 1.00 L of a buffer with a pH of 7.2 from 0.1 M
NaH2AsO4 solution and 0.12 M AsO43- solution?
6- Calculate the pH of a buffer solution that initially consists of 0.0400 moles of

NH3 and 0.0250 moles of NH4+ ion, after 20.0 ml of 0.75 M NaOH has been added
to the buffer. Ka for NH4+ ion is 5.6 x 10-10.
7- Calculate the pH of the following solutions?

a- 0.40 M formic acid solution (Ka = 1.7 10-4).
b- 0.25 M sodium benzoate solution (for benzoic acid, Ka = 6.5 10-5).
c- 0.32 M NH4Cl solution (for ammonia, Kb = 1.8 10-5).
d- 0.15 M NH3/0.35 M NH4Cl
e- 0.12 M HAc/0.20 M NaAc
f- 0.10 M Na2HPO4/0.15 M KH2PO4
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Experiment Six
Titration of Polyprotic acid
- Titration Curve of Phosphoric Acid H3PO4
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Experiment Six
Titration Curve of Phosphoric Acid H3PO4
I- Introduction:
Diprotic acids and bases are compounds that can donate or accept two
protons, while polyprotic acids and bases are compounds that can donate or
accept more than one proton. The followings are common polyprotic systems
with examples:
- Diprotic: Sulfuric Acid, Oxalic Acid.
- Triprotic: Phosphoric Acid.
- Amino Acids: Neutral molecule is a zwitter ion, a molecule that has both
a positive and negative charge. This form is due to the acidity of the
carboxylic acid functional group compared to that of the ammonium
group forcing the amino acid to rearrange to give the zwitter ion. At low
pH, both the ammonium and carboxyl groups are protonated, while at
high pH neither group is protonated. Protons are always lost (acid) or
gained (base) one at a time!
Each donation of a proton (acid) or acceptance of a proton (base) is

characterized by a unique Ka (or Kb) value. When you have a polyprotic acid or
base in solution you can have many different species present; however, depending
on the pH of the solution, one or two species is usually dominant. Some compounds
can function as either acids or bases depending on the situation. They are said to be
amphiprotic. Water is an example:
HC2H3O2 + H2O = H3O+ + C2H3O2-

Acid Base Acid Base
NH3 + H2O = NH4+ + OH-

Base Acid Acid Base
Amphiprotic molecules are typically the intermediates of a polyprotic acid or base.
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II- Purpose:
1- To draw the titration curve of H 3PO4 (pH versus ml NaOH addition).
2- To determine from the titration curve the pKa1, pKa2 and pKa3 of H 3PO4.
3- To determine the pH values at the first equivalent point and second equivalent
point.
III Theory:
- When an acid has more than one ionizable proton, the protons are lost in
successive reactions.
- Each ionization is characterized by a separate Ka value.
- Each successive Ka is smaller than the preceding one.
- For a weak polyprotic acid, the first ionization produces much of the H+ ions.
For the triprotic acid H 3PO4, three equilibrium reactions exist in the
aqueous solution:
H3PO4 + H2 O H3 O+ + H2PO4-
[H 2O ][H 2 PO4 ]
Ka1K1 7.1103 mol.L1
[H 3PO4 ]
H2PO4- + H2 O H3 O+ + HPO4 2-
2
Ka2K1 [ H 2O ][ HPO4 ] 6.2 10 8 mol.L1
[ H 2 PO4 ]
HPO42- + H2 O H3 O+ + PO43-
3
Ka3K1 [H 3O ][PO4 ] 4.4 1013 mol.L1
[HPO24 ]
Experimentally, Ka1 Ka2 and Ka3 can be determined from the titration
curve that is drawn for the change of pH in a solution of H 3PO4 by the addition
of strong basic solution NaOH.
As the acid becomes weaker, the jump gets smaller. The phosphate jump is
very small where its pka3 is closed to kw.
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- For monoprotic acid: Following Henderson-Hasselbalch equation

pH = pKa + log([A-]/[HA]), when the pH > pKa, the basic species is the
dominant one, whereas at pH < pKa, the acidic species is the dominant one.
For monoprotic, A- is predominant when pH > pKa
For monoprotic, HA is predominant when pH < pKa
- For polyprotic acid: the reasoning is the same as monoprotic acid, except
here there are multiple pKa values to separate different dominant species.
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What are the rules for Theoretical Calculation of pH of different species of

phosphoric acid:
H3A is treated as a monoprotic acid, Ka = K1
H2A- is treated similarly as an intermediate form of a diprotic acid
K 2 K1F K1 K w
[H ]
K1 F
The calculated pH should be closed to 1/2(pk1+pk2). Then [H+] is equal to the
square root of ka1.ka2.
Prove that H2PO4- is an amphoteric species that is more like an acid.
H2PO4- + H2 O OH- + H3PO4 kb3= k w/k a1
H2PO4- + H2 O H3 O+ + HPO42- ka1
Compare the values of ka1 and kb3
HA2- is also treated similarly as an intermediate form of a diprotic acid
- Surrounded by H2A- and A3-
- Use K2 & K3, instead of K1 & K2
K 2 K3 F K 2 K w
[H ]
K2 F
The calculated pH should be closed to 1/2(pk 2+pk3). Then [H+] is equal to the
square root of ka2.ka3.
Prove that HPO42- is an amphoteric species that is more like a base.
HPO42- + H2 O OH- + H2PO4- kb2= k w/k a2
HPO42- + H2 O H3 O+ + PO43- ka3
Compare the values of ka3 and kb2
A3- is treated as monobasic, with Kb=Kb1=Kw/Ka3
IV- Procedure:
Fill your burette with 0.100 M NAOH solution. Pipet 25.00 ml of 0.100 M
H3PO4 solution in a 150 ml beaker. Put in the beaker a magnetic bar, and then put the
beaker on the magnetic stirrer. Insert the electrodes of the pH-meter in the beaker and
record the pH value after the addition of 2.00 ml increments of NaOH solution until
you have about 80 ml NaOH solution.
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Data Table
Volume of H3PO4 in the beaker in ml
Volume of Volume of Volume of

pH pH pH
NaOH added NaOH added NaOH added
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Questions
1- Draw the titration curve of the pH of H3PO4 versus ml NaOH added. Show
on the curve the pKa1, pKa2 and the pKa3; pH at first equivalent point and the
pH at second equivalent point.
2- Calculate the experimental values of pKa1, pKa2 and the pKa3. Compare the
above values to theoretical ones. Comment on the possible sources of error.
The third jump is very small and is not clarified on the curve.Why?
3- H2SO4 is a strong acid in its first ionization; but a weak acid in its second.
Calculate the pH of a 0.056 M solution of H2SO4.
4- Calculate the hydronium ion concentration and the pH of a 10 -2 M NaHA
solution. Given: The dissociation constants of H2A are Ka1 = 10-2 and Ka2 = 10-7.
5- Comment on the following:
- Practically, the third equivalence point will not appear in the titration curve.
- Magnification of pka errors is an important source of error in the experiment.
6- Complete the following:
At the first equivalence point: n OH- = n H3PO4
What do we have at the second equivalence point?
7- Look for the methods of analysis of Titration curves in the following cases:
A polyprotic base
A mixture of strong acid and weak polyprotic acid.
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Part II
EDTA Titrations
- Exp Seven: Determination of Total Water Hardness
Hardness
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Experiment Seven
EDTA Determination of Total Water Hardness
Hardness
I. General Introduction:
Any reagent which reacts with an analyte in a known ratio and with a large
equilibrium constant can potentially be used in a titration.
Complexation Titrations are based on the reaction of a metal ion with a
chemical agent to form a metal-ligand complex.
Complexation Titrations are essentially a Lewis acid-base reaction, in which an

electron pair is donated from one chemical to another. The ligands used in
complexometric titrations are also known as chelating agents. The equilibrium
constant for the reaction between a metal ion (M +n) and a chelating agent (L-P) is
known as a formation constant or stability constant.
EDTA or ethylene di-amine tetra-acetic acid is a novel molecule for complexing

metal ions. It is a polyprotic acid containing four carboxylic acid groups and two
amine groups with lone pair electrons. The classic structural formula is given
below. EDTA is synthesized on an industrial scale from ethylene diamine,
formaldehyde, and a source of cyanide (HCN or NaCN). On a worldwide basis over
100,000 metric tons are produced annually.
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The unusual property of EDTA is its ability to chelate or complex metal ions in
1:1 metal-to-EDTA complexes. The fully deprotonated form (all acidic hydrogens
removed) of EDTA binds to the metal ion. The equilibrium or formation constants
for most metals, especially the transition metals, are very large, hence the reactions
are shifted to the complex. Many of the reactions are pH dependent, especially the
weaker forming complexes with Ca+2 or Mg+2.
M+n + Y-4 MYn-4 Kf = (MYn-4)/(M+n)(Y-4)

Metal analysis can be done by titration with EDTA and the use of a metal ion
indicator.
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What are the uses of EDTA?
Because of its strong complexing ability for most metal ions, it is used in the
food industry as a sequestering agent. The complexing of the metal ion may
prevent further reactions, such as binding metals that are cofactors for enzymes, or
just remove a metallic taste, such as metal contamination added during processing.
Recent studies have shown that NaFeEDTA and Na2EDTA added to typical
iron fortification compounds in cereals increased the absorption of iron in adult
humans.
This same property allows EDTA use for incidents of lead poisoning by the
medical profession. The formation constant for Pb-EDTA complex is 1018.
Intravenous injection of Na2CaEDTA solution is given at 25 mg/kg body mass/day
over 6 hours for 5 days when blood lead levels go over 45 mg/dL. The Pb+2 ion
replaces the Ca+2 ion in the complex because the formation constant for the lead
complex is greater than the calcium complex.
Pb+2 + CaY-2 PbY-2 + Ca+2 K ~ 108

The five day limit is there to prevent Zn+2 depletion, since the Zn+2 ion
replaces the Ca+2 ion in the complex too. EDTA is added to store blood in blood
banks as an anticoagulant to bind Ca+2 ion.
II. Purpose:
1- To determine the concentrations of Ca2+ and Mg2+ ions in a commercial
sample of bottled mineral water or tap water, then to compare
experimental results with the concentrations of the metal ions claimed by
the manufacturer.
2- To find the total concentration of metal ions that can react with EDTA,
assuming that this equals the concentration of Ca2+ and Mg2+, the most
common multivalent metal ions in natural waters.
3- To analyze Ca2+ separately after precipitating Mg(OH)2 with strong base.
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III. THEORY:
Water hardness is an expression for the sum of the calcium and magnesium cations
concentration in a water sample. These cations form insoluble salts with soap, decreasing
soaps cleaning effectiveness. They also form hard water deposits in hot water heaters.
The standard way to express water hardness is in ppm CaCO3 which has the formula
weight of 100.1 g/mole.
An excellent way to determine water hardness is to perform a complexometric
titration using a chelating agent standard solution, EDTA, usually in the form of (Y4-).
Ca2+(aq) + Y4-(aq) CaY2-(aq)
Mg2+(aq) + Y4-(aq) MgY2-(aq
Due to steric hindrances, EDTA will complex with calcium and magnesium in a one-
to-one molar ratio.
The endpoint in this experiment will be determined using Erichrome black T indicator
at pH 10. Ca2+ ion first complexes with the indicator as CaIn+ which is wine red. As the
stronger ligand EDTA is added, the CaIn+ complex is replaced by the CaY2- complex
which is blue. The end point of titration is indicated by a sharp colour change from wine
red to blue.
The indicator imparts a red color to the solution while there are calcium and
magnesium ions that have not complexed with EDTA. Once the endpoint has been
reached and there is no more uncomplexed Ca2+ or Mg2+, the indicator will give a blue
color. No hint of red color will be left.
Titration using Eriochrome Black T as indicator determines total hardness due to

Ca and Mg2+ ions.
2+
Hardness due to Ca2+ ion is determined by a separate titration at a higher pH, by adding
NaOH solution to precipitate Mg(OH)2(s), using hydroxynaphthol blue as indicator.
Labels used to describe water hardness can be notified from this table:
Concentration (mg CaCO3) Description

0-50 Soft
50-100 moderately hard
100-300 Hard
> 300 very hard
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IV- Materials and Apparatus
- Commercial sample of bottled mineral water or tap wate

- Eriochrome Black T and hydroxynaphthol blue indicators
- (NH4OH-NH4Cl) buffer pH 10
- 0.01 M EDTA (disodium salt)
- 50% w/v NaOH solution (50 g in 100 ml solution)
- measuring cylinder and the apparatus required for titration
V- Experimental Procedures:
You will be using the disodium salt of EDTA (M.W = 372.24 g/mole). It has been
dried for week at 80C to drive off any superficial moisture. It is in the TA desiccator. Be
sure to return it to the desiccator when you are through with it.
Weigh carefully about 0.9 g of EDTA (record to the nearest 0.1 mg). Quantitatively
transfer this into a 250 ml volumetric flask then add 2-3 ml of pH 10 ammonia buffer. Fill
the flask about halfway to the mark with distilled water and swirl to dissolve. This process
can take up to 15 minutes. Once dissolved, dilute to the mark and then cap and invert the
flask at least 6 times to get a uniform solution. Keep the solution capped.
Part A: Determination of total hardness

1. Pipette 50ml mineral water into an Erlenmeyer flask.
2. Add 2ml buffer solution followed by 3 drops of Eriochrome Black T indicators.
3. Titrate with 0.01 M EDTA until the solution turns from wine red to sky blue with no
hint of red (save the solution for colour comparison).
4. Repeat the titration to obtain two concordant results.
Part B: Determination of concentration of Ca2+(aq) ions

1. Pipette 50 ml of mineral water into an Erlenmeyer flask.
2. Add 30 drops of 50% w/v NaOH solution, swirl the solution and wait for a
couple of minutes to completely precipitate the magnesium ions as Mg(OH)2(s).
3. Add a pinch of hydroxynaphthol blue (exact amount to be decided by the
intensity of the resulting coloured solution) and titrate with 0.01 M EDTA until it changes
to sky blue (save the solution for colour comparison).
4. Repeat the titration to obtain two concordant results.
Caution:
- This lab will be graded primarily on the accuracy of your individual results. Due to the fact
that you will be using standardized EDTA as a primary standard, it is important that you be
extremely careful in your weighing procedure. Any mistakes will carry through the entire
experiment and greatly affect the accuracy
Data of your results. Careful titrations will give you
Tables
high precision and accuracy.
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Part A: Determination of total hardness

Trial 1 2 3
Initial burette reading /ml
Final burette reading/ ml
volume of 0.01 M EDTA used/ml
Average volume of EDTA
Part B: Determination of concentration of Ca2+ ions

Trial 1 2 3
Initial burette reading /ml
Final burette reading/ ml
volume of 0.01 M EDTA used/ml
Average volume of EDTA
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Questions
1. From the results in Part A, determine the total concentration of Ca2+ and Mg2+
ions in the mineral water sample in ppm.
2. From the results in Part B, determine the concentration of Ca2+ ions in the
mineral water sample in ppm.
3. Hence, calculate the concentration of Mg2+ ions in the mineral water
sample in ppm. Compare with the corresponding values displayed on the label
of the bottle.
4. Why are two indicators used in the experiment? Can the first indicator be
used for the second titration?
5. What are the limitations of the EDTA titration in determining metal ion
concentrations?
6. What is the effect of using wet EDTA instead of dried?
7. What is meant by soft water? Why we should add more buffer in analysis of soft
water?
8. A 50.0ml solution containing Ni2+ and Zn2+ was treated with 25.0 ml 0.0452M
EDTA to bind all the metal.The excess unreacted EDTA required 12.4 ml
0.0123M Mg2+ for complete reaction. An excess reagent is added again to replace
EDTA from Zn2+. Another 29.2 ml of Mg2+ wre required for ereaction with yhe
liberated EDTA. Calculate the molarity of Ni2+ and Zn2+ in the original solution.
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Part Three
Redox Titrations
- Exp Eight: Vitamin C Analysis
- Exp Nine: Bleach Analysis
- Exp Ten: Milk Analysis
- Exp Eleven: Electrochemistry
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Redox titration (also called oxidation-reduction titration) is a type of

titration based on a redox reaction between the analyte and titrant. Redox titration
may involve the use of a redox indicator and/or a potentiometer. There are a lot of
redox titrations classified according to the titrant used.
1) Permanganimetric: Titrant KMnO 4

2) Dichromatometric: Titrant K 2Cr2O7
3) Titrations involving iodine (I 2)
- Iodimetry
- Iodometry
In redox titrations a reducing agent is the element or compound in a redox
reaction that reduces another species. In doing so, it becomes oxidized, and is
therefore the electron donor in the redox. Examples of reducing agents: The
active metals sodium, magnesium, aluminum, and zinc.
An oxidaizing agent is the element or compound in a redox reaction that
oxidaizes another species. In doing so, it becomes reduced, and is therefore the
element or compund that gain electrons. Examples: permanganate (MnO 4),
chromate (CrO42-), and dichromate (Cr 2O72-) ions, sodium hypochlorite (bleach)
as well as nitric acid (HNO 3), perchloric acid (HClO 4), and sulfuric acid
(H2SO4).
How do you balance a redox reaction?

Just follow these steps!
Step 1: Write two unbalanced half-reactions, one for the species that is being oxidized
and its product, and one for the species that is reduced and its product.
Consider the example: IO3- + I- I2
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Here is the unbalanced half-reaction involving IO3- : IO3- I2
And the unbalanced half-reaction for I- is: I- I2
Step 2: Insert coefficients to make the numbers of atoms of all elements except oxygen
and hydrogen equal on the two sides of each half-reaction.
2IO3- I2
2I- I2
Step 3: Balance oxygen by adding H2O to one side of each half-reaction.

2IO3- I2 + 6H2O
2I- I2
Step 4: Balance hydrogen atoms. This is done differently for acidic versus basic solutions.
For acidic solutions: Add H+ to each side of each half-reaction that is "deficient" in
hydrogen (the side that has fewer H's) and add an equal amount of H2O to the other side.
For basic solutions: add H2O to the side of the half-reaction that is "deficient" in hydrogen
and add an equal amount of OH- to the other side.
Note that this step does not disrupt the oxygen balance from Step 3. In the example here, it
is in acidic solution, and so we have:
2IO3- + 12H+ I2 + 6H2O

2I- I2
Step 5: Balance charge by inserting e- (electrons) as a reactant or product in each half-

reaction.
2IO3- + 12H + + 10e- I2 + 6H2O
2I- I2 +2e-
Step 6: Multiply the two half-reactions by numbers chosen to make the number of
electrons given off by the oxidation step equal to the number taken up by the reduction
step. Then add the two half-reactions. If done correctly, the electrons should cancel out.
Here the oxidation half-reaction must be multiplied by 2 :
2IO3- + 12H+ + 4e- I2 + 6H2O
10I- 5I2 +4e-
The totat reaction is then: 2IO3- + 10I- +12H+ - 6I2 + 6H2O
We can divide by two: IO3- + 5I- +6H+ - 3I2 + 3H2O
Titrations that create or consume I2 are widely used in quantitative analysis.
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When a reducing analyte is titrated with iodine (the titrant), the method is
called iodimetry. It is a direct titration with only 1 reaction:
analyte (unknown) + titrant (known I 2) product (iodide I -)
Example, the quantification of Ascorbic Acid (vitamin C)

C6H8O6 + I2 C6H6O6 + 2I- +2H+
Iodine rapidly oxidizes ascorbic acid C 6H8O6 to produce dehydroxyascorbic

acid C 6H6O6.
Iodometry is the titration of iodine (I 2) produced when an oxidizing analyte is

added to excess I -(iodide).Then the iodine (I 2) is usually titrated with standard
thiosulfate solution. It is not a direct titration because there are 2 reactions:
analyte (unknown) + I- I2
I2 + titrant (Known standard thiosulfate) product
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Exp Eight
Determination of Vitamin C in Tablets and Juices
Ascorbic Acid or Vitamin C is the analyte of this experiment. It is the most

popular single vitamin. Besides taking it to treat colds, people pop vitamin C
capsules hoping that it will cure numerous ailments. There is now scientific
evidence to support some of that hope.
Scientifically controlled studies using vitamin C for colds show that it can
reduce the severity of cold symptoms, acting as a natural antihistamine. The vitamin
may be useful for allergy control for the same reason: It may reduce histamine
levels. By giving the immune system one of the important nutrients it needs, extra
vitamin C can often shorten the duration of the cold as well.
Vitamin C makes the headlines when it comes to cancer prevention. Its
antioxidant properties protect cells and their DNA from damage and mutation.
As an antioxidant, vitamin C helps to prevent cataracts -- the clouding of the lens of
the eye that can lead to blindness in older adults. The lens needs a lot of vitamin C
to counteract all the free radicals that form as a result of sunlight on the eye.
As with the other antioxidants, vitamin C helps to prevent heart disease by
preventing free radicals from damaging artery walls, which could lead to plaque
formation.People with diabetes can benefit from extra vitamin C, too. This nutrient
can help regulate blood sugar levels.
II- Purpose:
1- To practice redox titration
2- To determine the amount of Vit.C present in a Vit C tablet or Juice.
III- Theory:
Rather than titrating directly with iodine, a known excess of iodine will
be generated directly in the solution with the ascorbic acid, according to the
following reaction:
IO3- + 5l- + 6H+ 3I2 + 3H2O
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Vitamin C is readily oxidized by iodine in an acidic solution:
Since the standard solution lacks stability, a back titration is to be run. The excess
iodine which did not react with the ascorbic acid will then be back-titrated with
standard sodium thiosulfate solution. Thiosulfhate is a strong reducing agent used to
determine oxidizing agents indirectly; it is then oxidized into tetrathionate:
2S2O32- + I2 S4O62- + 2I-
Iodine forms a complex with starch which is dark blue and the endpoint
of the titration can be detected by the disappearance of this color. By knowing
the total quantity of iodine formed, and the quantity left after reaction with
vitamin C, the amount of iodine reacted with the vitamin C can be calculated.
Starch should be freshly prepared to avoid attach of microorganisms and
fungi. To avoid its hydrolysis in presence of iodine, it should be added near the
endpoint, when the solution turns straw yellow, denoting that most amount of
iodine has been reduced.
III- Chemicals:
* Starch solution 5 %
* Potassium iodate, KlO 3 : dried approximately at 100 oC for 1 hr and
completely cooled in the desiccators.
* Potassium iodide, KI: solid
* Sulfuric acid, H2SO40.2M
* Sodium carbonate, Na 2CO3 and sodium bicarbonate N HCO3
* Vitamin C tablets: 500mg Ascorbic Acid
* Sodium thiosulfate, Na 2S2O3: Boil 500 ml water for 5 min. Weigh sufficient
Na2S2O35H2O to make a 0.08M solution and add to the boiled water. Add 35 mg
Na2CO3 and store the solution in the dark in a 1liter amber bottle. This solution
will not be stable for more than one week.
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IV- Procedure:
Standardization of Na2S2O3 Titrant:
1- Weigh out precisely about 5.00g of KlO3. Transfer to a 250.0 ml volumetric
flask, add about 150 ml distilled water, mix well until the powder dissolves,
then complete to the mark.
2- Pipet 25.00 ml of KlO3 solution into a 100 ml Erlenmeyer flask and add
about 0.1 g of NaHCO3.
3- Add about 1 g KI and 10 ml of 0.2 M H2SO4. Fill the buret with Na 2S203
solution and slowly titrate until the solution is pale yellow. At this point, add 10
drops of starch solution and continue adding titrant drop wise until the dark
color just disappears.
3- Repeat step 2, two more times.
Determination of Vitamin C in a Tablet:

1- Weigh a vitamin C tablet and grind to a powder with a mortar and pestle.
2- Dissolve 0.1 g powder in about 15 ml distilled water using 100 ml
Erlenmeyer flask; Note that some additives remain insoluble.
4- Pipit 30.0 ml of the KlO 3 solution into the Erlenmeyer flask, add about 1 g of
KI, 10.0 ml of 0.2M H2SO4, and about 0.1 g of NaHCO3.
3- Stir to ensure that all of the KI dissolves. The solution should become a red
brown color due to the presence of I3- ion.
4-Begin titrating the solution with the thiosulfate solution. The redbrown color
should fade as the I3- is consumed. When the solution takes on a yellow color, add
10 dropsl starch indicator. The solution should turn a deep blue, or possibly a
greenbrown. Continue titrating until the blue color has disappeared. Record the
amount of thiosulfate used. Be sure to wait about 30 seconds after the color
disappears to ensure completion otherwise add a few drops of the Na2S2O3 solution.
5- Repeat the titration one more time, or as needed to ensure precision, adding 2
ml of starch solution as before just prior to the endpoint.
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Determination of Vitamin C in Juice
1- Use a graduated cylinder to measure out a 25.00 ml sample of either pineapple or

grapefruit juice. Add the juice to a 250 ml Erlenmeyer flask, and then add 20 ml
of 0.6 MH2SO4.
2- Add 1.00 g of KI and 25.00 ml of 0.02M KIO3 to the flask. Begin titrating with
the thiosulfate solution. As before, add the starch solution when the redbrown
color fades and continue titrating until the blue color disappears. However, note
that the solution will not turn completely clear, due to the color of the juice. You
will have to make your best estimate as to when to stop the titration.
3- Repeat the experiment a second time.
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Data Tables
Weight of KlO3 _______ g Milliomoles of KlO3______________
Table 1: Standardization of Na2S2O3

Replicate 1 2 3
Milliomoles I2 produced
Milliomoles Na2S2O3 required
Titration volume, ml
Na2S2O3 concentration
Average Na2S2O3 concentration ________ M
Table 2: Back titration of excess I2 after reaction with vitamin C

Replicate 1 2 3
Titation volume, ml
Milliomoles I2
Milliomoles I2 titrated
Milliomoles I2 produced
Milliomoles ascorbic acid
mg/tablet ascorbic acid
Average mg/tablet ascorbic acid: _______

% Ascorbic acid: ___________________
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Questions
1- Calculate the molarity of the standard KIO3 solution.
2- Calculate the molarity of the Na 2S2O3 from the concentration of the KIO3
solution and the volume of titrant needed to titrate 25.00 ml, of the
standard KIO3. Report your values for the concentration of Na 2S2O3, and
the mean, standard deviation, and relative standard deviation of these
results.
3- Calculate the molarity of ascorbic acid in each of your sample solutions
from the volume of Na 2S2O3 titrant required to titrate the excess iodine,
knowing the total amount of KlO 3 which was added, and using the average
molarity of the Na 2S2O3 titrant determined in step 2.
4- From each of your 3 determinations, calculate the amount (milligrams)

of ascorbic acid per tablet. Report the average mg/tablet, and the
necessary statistical parameters.
5- Sodium thiosulfate is not a primary standard. Why?

6- Sodium thiosulfate decomposes in an acidic environment. Write a balanced
equation for this reaction.
7- Do you have to dry potassium iodide before using it? Why (not)?
8- Why is sodium carbonate used in the preparation of the sodium thiosulfate
solution?
9- Why it is better to add oxalic acid to the stock solution of the vitamin C?
10- A 25.00ml sample of orange drink with 2 g of KI and 25.00 ml of 0.0130
M KlO3 added was back-titrated with a 0.0311 M sodium thiosulfate
solution. The titration took 31.24 ml of thiosulfate. Calculate the ascorbic
acid concentration in the orange drink in mg/ml. mwt of ascorbic
acid=176.12g/mole)
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Exp Nine
The strength of Laundry Bleach
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Exp Nine
The strength of Laundry Bleach
I- Introduction:
Bleach is an additive which aids detergents in the removal of soil and stains.
Through a process of oxidation, bleach changes the soil into soluble particles which
can more easily be washed away helping to whiten and brighten washable fabrics.
There are two types of bleach:
Chlorine bleach (common household bleach or sodium hypochlorite):

Chlorine bleach is a 5.25% solution of sodium hypochloride. It is quite
powerful and must be diluted with water for safe use on some fabrics. While a dry
form is available, the liquid version is the most common.
When chlorine bleach is used in washing laundry, the chemical ingredient

oxidizes helping to remove soil and organic matter. It acts as a disinfectant on
bacteria and viruses and generally whitens fabrics.
Oxygen bleach (all fabric bleach):

Oxygen-based or All-Fabric Bleach works more slowly than chlorine bleach
and contains sodium perborate, sodium precarbonate or hydrogen peroxide. It is
safe for use on almost all washable fabrics and colors.
When this bleach is used in the wash, the chemical ingredient oxidizes to help
remove soil and organic matter brightening the fabric and removing stains.
II - Purpose:
In this experiment, you will use an oxidation-reduction titration to determine
the quantity of NaOCl in commercial bleach.
III- Theory:
Hypochlorous acid (HClO) is one of the important chlorine oxoacids.
Solutions of sodium hypochlorite (NaOCl), a salt of that acid, are sold as laundry
bleach. The hypochlorite anion (ClO-) is a strong oxidizing agent.
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As you will see, you can determine the quantity of the ClO - ion in a solution
through two oxidation-reduction reactions.
First, a known quantity of this anion is reduced to Cl- ions in an acidic
solution, using excess potassium iodide. The I- ions are oxidized to I2 in this
reaction. The solution that results is brown because that is the color of I2 in water.
2H+ + OCl- + 2I- I2 + Cl- + H20
Second, the stochiometric amount of I2 is reduced once again to I- during a

titration with a solution of sodium thiosulfate. Thiosulfate anions (S2O32-) are
oxidized to tetrathionate ions (S4O62-) in this reaction.
I2 + 2S2032- 2I- + S4062

Although you could use the disappearance of the color due to aqueous I 2 to
detect the endpoint of the titration, this technique would not be very sensitive.
Instead, you will use starch as an indicator. Starch reacts with I 2 to form a dark blue
complex. Starch should be freshly prepared to avoid attach of microorganisms
and fungi. To avoid its hydrolysis in presence of iodine, it should be added near
the endpoint, when the solution turns straw yellow, denoting that most amount
of iodine has been reduced.
I2 + starch (starch-12 complex)
This reaction is reversible. Consequently, the blue color fades during the
course of the titration as I2 is consumed.
The endpoint occurs when one drop of the Na2S2O3 solution causes the color
to change from blue to colorless. A trial titration will allow you to locate the
endpoints of subsequent titrations more easily.
IV- Chemicals:
* Starch solution 0.2%
* Potassium iodide, KI
* Hydrochloric Acid, HCl 2 M
* Sodium carbonate, Na 2CO3
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* Bleach sample
* Sodium thiosulfate, Na 2S2O3: Boil 500 ml water for 5 min. Weigh
sufficient Na2S2O35H2O to make a 0.025M solution and add to the boiled
water. Add 15 mg Na2CO3 and store the solution in the dark in a 1liter
amber bottle. This solution will not be stable for more than one week.
V- Procedure:
1- Obtain about 70 ml of a 0.0250 M Na2S2O3 solution and 3 samples of solid KI.
Each of these samples should have a volume of about 1cm3. They can be stored
on pieces of waxed paper.
2- A buret containing the bleach solution will be available for general use. This
solution must be diluted, however. Record the initial buret reading to the nearest
0.01 ml. Carefully add about 3 ml of the solution to a 100ml volumetric flask.
Record the final buret reading to the nearest 0.01 ml, and calculate the volume of
undiluted bleach used.
3- Add distilled water to the flask until the bottom of the meniscus coincides with the
etched line on the flask. Add the last 0.5 ml with a medicine dropper. Insert a
stopper in the flask and mix the solution thoroughly.
CAUTION: Be careful in your handling of the solutions used in this

experiment. Undiluted bleach and hydrochloric acid can cause chemical burns and
ruin your clothes. In addition, bleach can be especially irritating to the eyes. If you
spill either of these solutions on you, wash the contaminated area thoroughly and
report the incident to your laboratory instructor. You may require further
treatment.
Cleaning and filling your buret

4- Instructions for using a buret can be found in the Introduction to this manual. Clean
your buret according to the directions and fill it with some of the Na2S2O3 solution.
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Doing the trial titration

1- Pipet 10.0 ml of the diluted bleach solution into a clean 125ml Erlenmeyer flask.
2- Add one of the samples of solid KI to the flask, followed by 20 ml of distilled
water from a graduated cylinder and 20 drops of 2 M HCl. Swirl to obtain a
homogeneous solution.
3- Record the initial buret reading to the closest 0.01 ml.
4- Place the flask under the buret with the capillary tip inside the mouth of the flask.
Insert a piece of white paper under the flask.
5- While swirling, add increments of about 1 ml of the Na2S2O3 solution to the flask.
Continue until the brown color fades to yellow.
6- Add 2ml of a 0.2% starch solution.
7- Continue to add 1ml portions of the Na2S2O3 solution until one addition causes the
solution to become colorless.
8- Record the final buret reading to the nearest 0.01 ml, and calculate the approximate
volume of the Na2S2O3 solution required for the titration.
Doing the exact titrations

1- Repeat Steps 1 through 4 of the procedures for the trial titration.
2- Subtract 2 ml from the volume found in the trial titration. Rapidly add this new
volume to the flask from the buret.
3- Add 2ml of the 0.2% starch indicator.
4- Rinse the walls of the flask with distilled water from a plastic wash bottle.
5- Continue the titration on a drop-by-drop basis. Swirl the flask rapidly after each
addition. Remember that the endpoint will occur when one drop results in a
colorless solution.
6- Record the final buret reading to the nearest 0.01 ml.
7- Repeat the procedure with a second sample of the diluted bleach.
8- If the volumes at the endpoints differ by more than 0.15 ml (about 3 drops) or
some other amount specified by your instructor, repeat the titrations with
additional samples of the diluted bleach until the required precision is obtained.
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Data Tables
Trial Trial Exact Exact
Titration (1) Titration (2)
Volume of diluted bleach used (ml)
Final buret reading (ml)
Initial buret reading (ml)
Volume of 0.025M Na2S2O3 (ml)
Moles of Na2S2O3
Moles of NaOCl
Molarity of NaOCl in diluted bleach (M)
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Questions
1- Calculate and record the molarity of the diluted bleach solution.

2- Calculate the molarity of the undiluted bleach solution.
3- Commercial bleaches such as Clorox are said to contain 5.25% NaOCl by
weight. Is this correct? Use your results and a density of 1.0 g/ml for the bleach
in your calculations. These calculations should reflect the precision of the
assumed density.
4- A 10.0ml sample of aqueous NaOCl is treated with excess KI in an acidic
solution. The quantity of iodine that is liberated is such that 27.46 ml of 0.0250 M
Na2S2O3 solution must be added to cause the disappearance of the dark blue color
due to the starch indicator. What is the molarity of the solution of NaOCl?
5- What safety precaution is cited in this experiment?
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Exp Ten
Determination of calcium by Oxalate (Calcium in Milk)
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Exp Ten
Determination of calcium by Oxalate (Calcium in Milk)
I- Introduction:
Calcium constitutes an important electrolyte in the human body, having an

approximate concentration of 10 mg per 100 cm of body fluids. It decreases the
permeability and irritability of cells, it is essential to bone and teeth structures, to
muscle contractibility, and is a necessary, but as yet poorly understood,
requirement for blood coagulation.
But the surprising benefits of calcium we dont know, that calcium protects our
health in other important ways, too. Here's what this amazing mineral can do:
Calcium helps keep the weight off: Research suggests that if you don't get
enough calcium in your diet, you're likely to be overweight,
Calcium protects your heart: If you're low on calcium, researchers say, you're
more likely to have high blood pressure.
Calcium improves premenstrual moods: A 1998 study led by Susan Thys-
Jacobs, M.D., of St. Luke's Roosevelt Hospital in New York City, found that
getting enough calcium can ease the symptoms of premenstrual syndrome
(PMS).
Calcium protects against colon cancer: Adequate calcium intake may reduce
your overall risk of colon cancer and suppress the growth of polyps that can
lead to cancer.
Calcium maintains healthy teeth: Calcium protects your teeth in an indirect
way. Your teeth themselves are relatively inert, meaning that the calcium
they contain usually stays there.
Milk products are the best source of calcium in the world. Not only do
milk products contain substantial quantities of calcium, but also the
calcium they contain is readily absorbed by the body.
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II- Purpose:
To learn how one can determine the amount of calcium, whether in milk,
eggs shells, bone or teeth. The procedure involves studying two
experimental techniques.
1- Precipitation of calcium by the oxalate.
2- Determination of calcium by red-ox titration.
III- Theory:
One of the oldest reactions known for the determination of calcium is via its
complexation with oxalate.
A complex, such as calcium oxalate, is a compound in which a metal cation
Ca2+ in this case, is surrounded by and bonded to one or several molecules or
anions called ligands. Complex compounds are essential to much biological
system, the most important of these compounds being chlorophyll and
hemoglobin.
Some ligands have several bonding sites, as is the case with oxalate, which
has two points of attachment.
The first step in the procedure involves the precipitation of calcium with
oxalate, C2O42-:
Ca2+ + C2O42- CaC2O4(s)
It is neither convenient nor accurate to weigh this precipitate directly.
Therefore, the calcium oxalate precipitate, one formed, is filtered, washed
thoroughly to remove any excess C 2O42- and then reacted with strong acid to
remove oxalic acid H2C2O4:
CaC2 O4(s) + 2H2SO4 Ca2+ + 2HSO4- + H2C2O4.
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The oxalic acid solution, the amount of which is a measure of the amount
of Ca 2+ initially present, is then titrated with potassium permanganate KMnO 4,
according to the following equation.
2KMnO4 + 5H2C2O4 + 3H2SO4 K2SO4 + 2MnSO4 + 10CO2 + 8H2O.
V- Procedure:
Caution: the ammonium oxalate solution is a toxic solution. You should

wash your hands carefully before leaving the laboratory.
1- Into 150 ml beaker, carefully measure 15 ml of the milk sample, using a

graduated cylinder.
2- To this solution add approximately (using a small graduated cylinder) 7 ml
of saturated ammonium oxalate (NH 4)2C2 O4, forming the insoluble white
complex.
3- Heat the mixture in water bath at about 60C for 15 minutes to allow the
precipitation process to come to completion.
4- Cool and then separate the white precipitate from the solution by
centrifugation, distributing the mixture equally into three or four centrifuge
tubes. Make sure that the test tubes are distributed symmetrically in the
centrifuge to ensure spinning.
5- After centrifugation, remove the tubes and decant (pour out) the supernatant
(to liquid).
6- Add about 3 ml of 2% ammonia solution to each tube, and stir until the white
precipitate mixes thoroughly with the ammonia. Stirring may be done with a
small stirring rod, but when finished the rod must be washed off with a little
distilled water and the washings combined with the solution. Remember, this is
a quantitative experiment.
7- When mixing is completed, centrifuge and decant as before.
8- Transfer the white precipitate from all tubes into 125 ml Erlenmeyer flask as
well. Use some amount of distilled water to aid the transfer. Wash the tubes
several time to transfer all the residue to the Erlenmeyer flask.
9- Add enough water to the Erlenmeyer flask to make a total of 50 ml.
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10- Add 10 ml of 6 mol.L -1 H2SO4, which should cause the entire precipitate to
dissolve,
11- Place the flask into the boiling water for a few minutes. Milk samples will
become cloudy.
12- Thoroughly clean a burette. Rinse it with some 0.02 mol.L-1 KMnO4
solution, and then fill the burette with 0.02 mol.L-I KMnO4 Solution. Record
the initial reading of the burette.
13- Place the flask containing the hot oxalate solution under the burette tip and
onto a white sheet of paper. Titrate the solution very slowly (any mistake is
repetition from the beginning), while constantly swirling the flask. At first
the permanganate solution (purple) will be decolorized immediately upon
coming in contact with the oxalate solution, but after a while the purple color
will begin to remain longer. This is an indication of the upcoming end-point.
Reduce the flow of the permanganate adding. It drop wise until a light, yet
definite purple-pink color appears and remains for at least 30 seconds. Record
the volume of KMnO 4 used.
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Data Tables
Name of milk brand

Volume milk sample
Molar concentration of KMnO4 solution
Initial burette reading
Final burette reading
Volume of KMnO4 used
Questions
1- Calculate the number of milliomol of KMnO4 used.

2- Calculate the number of milliomol of calcium oxalate that was precipitated;
deduce the milliomol of calcium in the milk sample.
3- Find the weight of calcium in milk sample (per 15 ml sampe).
4- Report the amount of calcium in the milk sample as mg/100 ml. Compare your
value to theoretical ones if applicable , comment on % relative error.
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Exp Eleven
Electrochemistry
Hardness
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Experiment Eleven
Electrochemistry
Hardness
A drug ion may be distributed passively across a membrane in accordance

with the membrane potential, the charge on the drug ion, and other factors.
The relationship between the membrane potential to the passive distribution
of ions across the membrane can be expressed quantitatively by Nernst Equation.
This equation will be discussed throughout this experiment.
The area of Chemistry that deals with the inter-conversion of electrical
energy and chemical energy is Electrochemistry. Electrochemical processes are
redox reactions in which the energy released by a spontaneous reaction is converted
to electricity.
II- Purpose:
1- To get acquainted with voltage measurements (using a voltmeter).
2- To measure the cell potential of different galvanic cells.
3- To learn how to construct a concentration cell and measure its potential.
4- To compare the observed cell potentials with those calculated from literature
values of electrode potentials and the Nernst equation.
III- Theory:
Redox reactions consist of two half-reactions: the oxidation half-reaction
(loss of electrons) and the reduction half-reaction (gain of electrons).
A redox reaction occurs when the oxidizing agent is in contact with the
reducing agent. The electrons are transferred directly from the reducing agent to the
oxidizing agent in solution. If these agents are physically separated from one
another, the transfer of electrons can take place via an external conducting medium.
As the reaction progresses, it sets up a constant flow of electrons and hence
generates electricity.
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The experimental apparatus for generation of electricity through the use of a

redox reaction is called an electrochemical cell.
A galvanic cell is an electrochemical cell in which electricity is produced by
means of a spontaneous reaction. As an example, consider a zinc bar dipped into a
ZnSO4 solution, and a copper bar dipped into a CuSO4 solution. The bars are called
electrodes. By definition, the electrode at which oxidation occurs is called the
anode, and the electrode at which reduction occurs is called the cathode.
Therefore,
Zn electrode, anode: Zn(s) Zn2+ (aq) + 2e-
Cu electrode, cathode: Cu2+ (aq) + 2e- Cu(s)
Cu2+ (aq) + Zn (s) Zn2+ (aq) + Cu (s)
The two solutions must be separated from each other; otherwise, the Cu2+
ions will react directly with the zinc bar, and no useful electrical work will be
obtained. However, to complete the electric circuit, the solutions must be connected
by a conducting medium, a salt bridge, which, in its simplest form, in an inverted
U-tube containing an inert electrolyte such as KCl or NH4NO3 in agar. During the
reaction, electrons flow externally from the anode (Zn electrode) through the wire
and voltmeter to the cathode (Cu electrode). In the solution, the cations (Zn 2+, Cu2+
and K+) move toward the cathode, while the anions (SO 42- and Cl-) move in the
opposite direction, toward the anode.
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By convention, the negative electrode in a galvanic cell is the electrode

from which electrons are emitted (i.e. where oxidation occurs, anode). The
positive electrode is the electrode to which the electrons are transferred (i.e. where
reduction occurs, cathode). The negative electrode is connected to the minus pole
of the voltmeter; thus, the positive electrode is connected to the plus pole of the
voltmeter.
The fact that electrons flow from one electrode to the
other indicates that there is a voltage difference between the
two electrodes, called the electromotive force, or emf (E),
which can be measured by connecting both electrodes to a
voltmeter.
The emf of a galvanic cell, also referred to as cell
potential or cell voltage, is a direct measure of the driving force or thermodynamic
tendency of the spontaneous redox reaction to occur. The cell potential is the sum
of the two electrode potentials. It is not possible, therefore, to measure individual
absolute electrode potentials. However, a usual procedure is to assign a value of
0.00 Volts to the standard potential for the electrode reaction:
2H+ + 2e- H2(g) EH+,H2 = 0.000 V
then, other electrode potentials can be given definite values based on the
assignedvalue.
The cell potential depends on the nature of the electrodes
and the ions, the concentrations of the ions, and the temperature
at which the cell is operated. The conventional notation for
representing galvanic cells is the cell diagram. The cell diagram
for the cell just described is:
Zn(s) | ZnSO4(aq,0.100M) | KCl(sat'd) | CuSO 4(aq, 0.100M) | Cu(s)
where the vertical lines represent phase boundaries; the Zn bar is a solid and the
ZnSO4 is a solution, thus a line is drawn in between to show the phase boundary.
The concentration of the species in solution is given, and the salt bridge is
included in between two vertical lines. The anode is written first at the left (always
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by convention) and the other components appear in the order in which they are
encountered in moving from anode to cathode.
The Nernst Equation:

As mentioned earlier, the emf of a cell depends on the concentrations of
reactants and products. For a redox reaction of the type
aA + bB cC + dD
the relationship between emf and concentrations of the species is given by
RT
E cell E ocell ln Q (1)
nF
where Eocell is the emf of the cell under standard-state conditions, R is the
universal gas constant, T is the absolute temperature, n is the number of moles of
electrons transferred during the course of the reaction, F is Faraday's constant, and
Q is the reaction quotient, given by:
[C]c [D]d
Q (2)
[A]a [B]b
Equation (1) is known as the Nernst equation. At 298 0K (25o), the Nernst
equation can be written as:
0.02569 (3)
E cell E o
cell ln Q
n
IV- Procedure:
A- Chemicals:
1- Metal bars: Fe, Cu, Pb.
2- Solutions: FeSO4, CuSO4, Pb(NO3)2,
(all solutions at concentrations 0.100 M and 0.0010 M)
3- 50 ml beakers,
4- KNO3 salt bridge,
5- Voltmeter.
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B- Galvanic Cells:
1- Prepare the following Galvanic cell:
Fe(s) | FeSO4(0.100M) || CuSO4 (0.100M) | Cu(s)
Use 50 ml beakers for the solutions compartments, and Zn and Cu metal
bars for the electrodes. Obtain from the store room a KCl salt bridge
(polyethylene tube containing a 1.00 M KCl agar gel), and use it for the
connection of the two solutions.
Measure the cell potential. Compare the predicted cell potential from the
literature values of the two reduction potentials. Enter data in Table 1.
Repeat the same experiment by changing the concentrations of the two
solutions as follows:
0.100 M FeSO4 / 0.0010 M CuSO4,
then: 0.0010 M FeSO4 / 0.100 M CuSO4.
Compare with the cell potentials calculated from the Nernst equation.
Enter values in table 2.
Measure and record the temperature of the experiment.
2- Prepare the following cell:

Fe(s) | FeSO4 (0.100 M) || Pb(NO3)2 (0.100M) | Pb(s)
Repeat the procedure in (1) for this cell. Enter measurements in Table 2.
3- Prepare the following cell:

Pb(s) | Pb(NO3)2 (0.100 M) || CuSO4 (0.100 M) | Cu(s)
Repeat the procedure in (1) for this cell. Enter measurements in Table 2.
Concentration cells:
Select a metal M and construct the following concentration cell:
M(s) | M2+ (0.0010 M) || M2+ (0.10 M) | M(s)
Measure the cell potential. Compare with the cell potential calculated from
the Nernst equation. Enter values in Table 3.
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Data Tables
Table 1: Emf of Galvanic Cells @ T =
# Cell Diagram Ecell Ecell % Error

Measured Theoretical
1 Fe(s)|FeSO4(0.100M)||CuSO4(0.100M) | Cu(s)
2 Fe(s) |FeSO4 (0.100 M) || Pb(NO3)2 (0.100M | Pb(s)
3 Pb(s) |Pb(NO3)2(0.100M)|| CuSO4 (0.100 M) | Ag(s)
Table 2: Comparison of Measured Emf and Calculated Values from Nernst Equation @ T =
# Cell Diagram Ecell Ecell %

measured calculated Error
1a Fe(s) | FeSO4(0.100M) || CuSO4 (0.001M) | Cu(s)
1b Fe(s) | FeSO4(0.001M) || CuSO4 (0.100M) | Cu(s)
2a Fe(s) | FeSO4(0.100M) || Pb(NO3)2(0.001M) | Pb(s)
2b Fe(s) | FeSO4(0.001M) || Pb(NO3)2 (0.100M) | Pb(s)
3a Pb(s) | Pb(NO3)2(0.100M)|| CuSO4(0.001M) | Cu(s)
3b Pb(s) | Pb(NO3)2(0.001M)|| CuSO4(0.100M) | Cu(s)
Table 3: Emf of Concentration Cell
# Cell Diagram Ecell Ecell %

Measured Theoratical Error
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Questions
1- Show one sample calculation for all the previous tables.
2- Write-out a redox reaction that takes place spontaneously using the

Sn2+ / Sn and Cd2+ / Cd redox couples.
Calculate the standard cell potential; then write the cell notation.
Given: E oCd
2
/ Cd
0.40V; ESn
o
2
/ Sn
0.14V
3- Calculate Eo and E (the cell emf) for the following cell reaction:
a- Mg(s) + Sn2+(aq) Mg2+(aq) + Sn(s).
[Mg2+] = 0.045 M; [Sn2+] = 0.035 M.
b- 3Zn(s) + 2Cr3+(aq) 3Zn2+(aq) + 2Cr(s).

[Cr3+] = 0.010 M; [Zn2+] = 0.0085 M.
Given: E oZn 2
/ Zn
0.76 ; E oCr3 / Cr 0.74V ; E oNg2 / Mg 2.37V
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Part IV
Spectrophotometric Instrumental Analysis
Exp 12: Spectrophotometric Analysis of a Commercial Aspirin Tablet

Exp 13: Quantitative Analysis for a Mixture Two Ions by Visible Spectroscopy
Exp 14: The Formula of a Complex Ion: Mole Ratio Method
Exp 15: Determination of the Equilibrium Constant for a Chemical Reaction
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Spectrophotometry Beer-Lambert Law
Ultraviolet-visible spectroscopy or ultraviolet-visible spectrophotometry refers

to absorption spectroscopy in the ultraviolet-visible spectral region. This means it
uses light in the visible and adjacent (near-UV and near-infrared (NIR)) ranges. The
absorption in the visible range directly affects the perceived color of the chemicals
involved. In this region of the electromagnetic spectrum, molecules undergo
electronic transitions. This technique is complementary to fluorescence
spectroscopy, in that fluorescence deals with transitions from the excited state to the
ground state, while absorption measures transitions from the ground state to the
excited state.
The electromagnetic spectrum covers an immense range of wavelengths. If a

solution is colored, it is because it absorbs light at some wavelength(s) from the
visible region of the electromagnetic spectrum. This removes them from the
spectrum, and the solution will appear to be a mixture of the colors not absorbed.
For example, the iron(II)-o-phenanthroline complex absorbs green and blue light,
making it appear orange. You will be measuring how much light various solutions
absorb at 508 nm (a particular shade of green), and using those measurements to
calculate the concentration of iron(II)-o-phenanthroline in those solutions.
Spectrophotometry is the quantitative measurement of the reflection or

transmission properties of a material as a function of wavelength. While relatively
simple in concept, determining the reflectance or transmittance involves careful
consideration of the geometrical and spectral condition.
Spectrophotometry involves the use of a spectrophotometer. A

spectrophotometer is a photometer (a device for measuring light intensity) that can
measure intensity as a function of the color (or more specifically the wavelength) of
light. Important features of spectrophotometers are spectral bandwidth and linear
range of absorption measurement.
Different types of spectrophotometers are found in

the market. They are classified on various basis. The
most important distinctions used to classify them are:
1. The different measurement techniques.
2. The wavelengths they work with.
3. How they acquire a spectrum.
4. The sources of intensity variation.
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Principle of Spectrophotometry
A spectrophotometer, one of the most useful physics lab equipments is the

combination of two devices, a spectrometer and a photometer. Spectrometer is used
for producing light of any selected wavelength or color while a photometer is used
for measuring the intensity of light. The two devices are placed at either side of a
cuvette filled with a liquid. Spectrometer produces the light of desired wavelength
and it passes through the tube and reaches photometer that measures its intensity.
Then the photometer produces a voltage signal to a display device, usually a
galvanometer.
As the amount of light absorbed by the liquid changes the signal also changes.
The concentration of a substance in solution can be measured by calculating the
amount of absorption of light at the appropriate wavelength or a particular color.
This instrument is representative of instruments in which a degree of accuracy is

sacrificed in exchange for simplicity of operation and low cost. Its normal range is
350 to 650 nm although this can be extended to 900 nm by the use of a red-sensitive
phototube.
The principle of the spectrophotometry process applied here is the following:
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In 1852, Prof. August Beer postulated that the decrease in intensity (or power)
of a beam of monochromatic radiation (a single wavelength of light) was
proportional to the intensity of the beam and to the amount of absorbing substance
in its path.
Beer-Lambert Law is more commonly known as Beer's Law. It states that the
optical absorbance of a chromophore in a transparent solvent varies linearly with
both the sample cell pathlength and the chromophore concentration. Beer's Law is
the simple solution to the more general description of Maxwell's far-field equations
describing the interaction of light with matter.
In practice, Beer's Law is accurate enough for a range of chromophores, solvents

and concentrations, and is a widely used relationship in quantitative spectroscopy.
The relationship between the power or intensity of the original beam of light (I O)
and of the emerging beam of light (I) is referred to as transmittance (T):
T= I / I0
% T =( I / I0 )* 100%
Io = intensity or power of original beam of radiation

I = intensity remaining after beam of radiation has passed through sample
b = length of sample in centimeters
The relationship between solution concentration and transmittance is not direct;

however, transmittance is readily converted into absorbance (A):
log(1/T)=A
then A= -log T
When using percent transmittance (%T), these equations become:
log 100 log %T = A
then 2 log %T = A
Unlike transmittance, absorbance corresponds directly to concentration.

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This is Beers Lamberts Law:

A= log(I / I0)= bc
where is the absorptivity constant (called the molar absorptivity constant or the
extinction coefficient when c, the concentration of the absorbing species, is given
in moles/liters), and b is the cell width or path length (the distance the light must
travel to pass through the sample; generally expressed in centimeters).
Last Equation is the fundamental law governing the absorption of all types of
electromagnetic radiation. It applies to solids and gases as well as solutions.
Absorbance is measured in a spectrophotometer by passing a collimated beam of
light at wavelength through a plane parallel slab of material that is normal to the
beam. For liquids, the sample is held in an optically flat, transparent container
called a cuvette. Absorbance (A) is calculated from the ratio of light energy
passing through the sample (I0) to the energy that is incident on the sample (I).
In an absorbance experiment, light is attenuated not only by the chromophore,

but also by reflections from the interface between air and the sample, the sample
and the cuvette, and absorbance by the solvent. These factors can be quantified
separately, but are often removed by defining I0 as the light passing through a
sample "blank" or "baseline" or reference sample (for example, a cuvette filled with
solvent but zero concentration of the chromophore is used as the blank).
So, a blank is generally used in spectophotometry in order to correct for a loss in

beam power at each interface due to reflection and to scattering by large molecules.
The intensity of the beam passing through the solution of interest can therefore
be compared with the intensity of a beam passing through an identical cell
containing just solvent (or just solvent + additives). Practically speaking, this means
that, in the last equation, Io refers to the intensity of the beam passing through the
blank.
Many factors can affect the validity of Beer's Law. It

is usual to check for the linearity of Beer's Law for a
chromophore by measuring the absorbance of a series of
standards. This "calibration" can also remove errors in
the experiment, the equipment, and the batch of
reagents (such as cuvettes of unknown pathlength).
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A Diagram of BeerLambert absorption

of a beam of light as it travels through a
cuvette of width is presented here.
UV/Vis spectroscopy is routinely used

in the quantitative determination of
solutions of transition metal ions and highly
conjugated organic compounds.
Solutions of transition metal ions can

be colored (i.e., absorb visible light) because d electrons within the metal
atoms can be excited from one electronic state to another. The colour of
metal ion solutions is strongly affected by the presence of other species, such
as certain anions or ligands.
Organic compounds, especially those with a high degree of conjugation, also
absorb light in the UV or visible regions of the electromagnetic spectrum.
The solvents for these determinations are often water for water soluble
compounds, or ethanol for organic-soluble compounds. Solvent polarity and
pH can affect the absorption spectrum of an organic compound.
While charge transfer complexes also give rise to colours, the colours are
often too intense to be used for quantitative measurement.
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A Scheme to follow in spectrophoometric analysis of an unknown sample
Topic Concepts Experiment
A standard solution is
Standard Solutions The dilution principle is
prepared and serial dilutions
Preparations already known and studied.
are carried.
Absorbance Spectrum The absorbance spectrum is Using the most concentrated

obtained from the plot of dilution,
absorbance against a wide the absorbance spectrum for
range of wavelength. It is a sample is measured. Then
used to detect max. It is max is detected. It will be
described and its importance used for all absorbance
is explained. measurements throughout
the experiment.
calibration curve Beer's Law is presented and The absorbance values of

explained. the serial dilutions are
determined at max.
A calibration curve, the plot
of absorbance vs
concentration is prepared,
and the molar absorptivity is
determined from the slope of
the plot.
Analysis of an The construction of a The prepared calibration

Unknown Solution calibration curve is curve is used to determine
described, and the use of a the analyte concentration in
calibration curve in the unknown solution from
determining the analyte Beer's Law equation.
concentration in an
unknown solution is
explained.
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Exp 12
Spectrophotometric Analysis of a Commercial Aspirin
Tablet
I- Introduction
Acetyl salicylic acid (ASA) is one of the oldest synthetic drugs. First synthesized in
Germany by the Bayer Company and marketed under the name Aspirin it has
remained one of the most popular over the counter drugs of all time. Its main
effect is as a pain killer and fever depressant, but in addition there is strong
evidence that in low daily dosages it lowers the incidence of heart attacks. In the
last few decades other drugs such as acetaminophen (commercial trade name
Panadol, also Tylenol) and ibuprofen (trade name Advil) have taken much of the
market for ASA, but ASA remains an important and widely used medicine.
Drugs, in addition to their active compound, often contain other inactive ingredients
(called excipients in the pharmaceutical industry) such as binders, fillers, dyes,
drying agents, etc. The content of active ingredient in a tablet will always be stated
on the package. In this experiment we will determine the percent active compound
in a commercial aspirin tablet. Aspirin is the trade name for acetylsalicylic acid
(ASA). The ASA in the tablet will be reacted with Fe3+, forming an intensely violet
colored complex. The concentration of the complex will be determined by means of
spectrophotometry, using a UV/VIS spectrophotometer. Finally, we will be able to
calculate the weight and the weight% of ASA in the commercial tablet.
II- Purpose
After completing this experiment, the student should be able to:

1. Prepare standard solutions.
2. Construct calibration curve based on Beers Law.
3. Use Beers Law to determine molar absorptivity.
4. Explain the fundamental principal behind spectrophotometric analysis.
III- Theory
Acetylsalicylic acid is the acetate (ethanoate) ester of salicylic acid, 2-
hydroxybenzoic acid. The acetyl ester is rapidly hydrolyzed to the salicylate
anion in basic medium, as shown in the following reaction.
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Once the de-esterification is complete, the solution is acidified, and FeCl3 is added.
The salicylic acid will react with the Fe3+ to form a colored complex ion:
In order to calculate the concentration of the complex we would need to know the
value of and l. Instead, we will measure the absorbance of Fe-salicylate complex
solutions of known concentration, and plot the absorbances of a number of such
known solutions vs. the concentration. This is known as a calibration curve. The
calibration solutions are prepared by first making a solution of the Fe-salicylate
complex of known concentration. This solution is called the stock solution. Next we
make 5 standard solutions by diluting a known amount of stock solution. The
Absorbance of each of these 5 solutions is measured, and plotted vs. their
concentration resulting in a linear calibration curve of A vs C. Next we measure the
absorbance of the solution prepared from the commercial aspirin solution, and find
its concentration by comparing its absorbance value on the calibration curve.
IV- Safety precautions

1.0M NaOH is caustic. Avoid contact with skin, wear gloves and safety glasses at
all times. Be particularly careful when heating the ASA-NaOH mixture on the hot
plate, cover with watch glass to prevent spattering.
V- Materials and chemicals

UV/VIS spectrophotometer and polystyrene cuvettes, hot plate, 1x 10 mL graduated
cylinders, 2 x100 mL volumetric flask, 6 x25 mL volumetric flasks, 1 mL graduated
pipet or micropipette, 2 x125 mL Erlenmeyer flasks or 150 mL beakers, watch
glass.
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Salicylic acid (reagent grade), 1.0 M NaOH, 0.02M FeCl3 (buffered to pH = 1.6
with HCl/KCl) Commercial AspirinTM or ASA tablets for analysis (not to be used
for your headache caused by this experiment!).
VI- Disposal
The remaining NaOH and Fe-salicylate solutions can be combined and neutralized
before disposal.
VII- Procedure
1. Operation of the spectrophotometer.
Follow the instructions in the laboratory. Use FeCl3 as the blank solution to zero the
absorbance reading.
2. Preparation of the Fe-salicylate standard solutions and ASA unknown

The preparation and measurements of
the SA standards and ASA unknown
can be done simultaneously.
Salicylic Acid (SA) Aspirin (ASA)

1. Weigh approximately 0.160 g SA 1. Weigh ASA tablet in beaker,
directly into small dry beaker. Record record the mass, crush tablet.
the mass (g). 2. Add 5 mL NaOH.
2. Add 5 mL NaOH. 3. Heat on hot plate until dissolved
3. Heat on hot plate until dissolved. (some fine non-active ingredients
Cool. may still be visible). Cool.
4. Transfer solution to 100 mL 4. Transfer solution to 100 mL
volumetric flask. Fill to the mark with volumetric flask.
distilled water. Fill to the mark with distilled water.
This is your stock SA solution This is your stock ASA solution
5. Prepare 5 standard solutions in 25 5. Prepare one ASA solution: use a 1
mL volumetric flasks: use a 1 mL mL graduated pipet to add 0.75 mL
graduated pipet to add 0.25, 0.5, 0.75, ASA stock in 25 mL volumetric flask.
1, and 1.25 mL stock SA into 25 mL Fill the flask to the mark with
volumetric flasks. Fill each flask to acidified FeCl3solution.
the mark with acidified FeCl3solution. The solution will have a medium dark
Solutions will vary from light to dark violet colour.
violet colour. 6. Measure and record the absorbance
Mark the flasks 1 (0.25 mL) to 5 at 530 nm
(1.25 mL)
6. Measure and record the absorbance
of each solution at 530 nm, using the
FeCl3 solution as blank.
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Questions
Give detailed reasons for the following statements.
a. Measurements of absorbances of compounds are carried at max.

b. A blank or reference solution is usually used in spectrophotmetry.
c. The value of absorbance never exceeds 2.
d. Beer`s Law is better to be applied to diluted solutions.
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Exp 13
Quantitative Analysis for a Mixture Two Ions by Visible Spectroscopy
Exp 13
Quantitative Analysis for a Mixture113
Two Ions by Visible Spectroscopy
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When a beam of parallel radiation passes through a layer of solution having a

thickness, b (cm) and a concentration, c (moles per liter) of an absorbing species, a
consequence of interactions of the photons and the absorbing particles is attenuation
of the beam.
The transmittance (T) of the solution is the fraction of the 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 through the solution
and the concentration of the absorbing species. That is, A= bc where is a
proportionality constant called the molar absorptivity. has units of M -1 cm-1 when
b and c are expressed in cm and moles per liter respectively. This relationship
between absorbance (A) and bc is known as Beer's Law.
Beer's Law is successful in describing the absorption behavior of dilute
solutions only. At high concentrations, the average distance between the species
responsible for absorption is diminished to the point where each effects the charge
distribution of its neighbors. This interaction, in turn, can alter their ability to
absorb a given wavelength of radiation.
Because the extent of interaction depends upon concentration, the occurrence of
this phenomenon causes deviations from the linear relationship between absorbance
and concentration. A similar effect is sometimes observed in solutions containing
high concentrations of electrolytes. The proximity of ions (in addition to other
factors such as temperature) alters the molar absorptivity of the absorbing species.
So, Beer's law states that absorbance of electromagnetic radiation is directly
proportional to concentration: if there is more than one absorbing species in
solution, the total absorbance is the sum of the individual absorbances of all
the absorbing species, provided there is no interaction among the various
species. By measuring n absorbances at n different wavelengths, n different
absorbing components could theoretically be measured by this technique.
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II- Purpose:
To analyze a mixture of two ions by a visible spectroscopy.
III- Theory:
Analyzing a mixture of two ions does not necessarily involve separation of
the ion mixture before measurement. For example, a mixture of Ca 2+ and Mg2+ can
be analyzed by a complexometric titration; a mixture of Co2+ and Cr2+ can be
analyzed by spectroscopy, a mixture of Fe3+ and Al3+ can be analyzed by
precipitation.
In this experiment, you will quantitatively determine the concentrations of
permanganate (MnO4- ) and dichromate (Cr2O72-) ions in a mixture. Because of the
overlap of spectra, it is not feasible to generate a calibration curve of one ion and
then the other and use a straightforward Beers Law approach to determine each
concentration. The problem is that each ion has significant absorbance at the others
max. The spectrum for Cr 2O72- possesses a maximum at 440 nm whilst MnO 4-
possesses a maximum at 545 nm and another at 525 nm. The analysis will be
carried out at 545 nm where there is relatively less interference. These
wavelengths correspond, more or less, to wavelengths where the absorbance is not
changing much over a range of wavelengths. Hence, small deviations in the
measurement wavelength do not affect the measured absorbance.
So, a mixture of the two cations, Mn7+ and Cr6+ can be analyzed by visible
spectroscopy after having oxidized the two ions to MnO 4- and Cr2O72- by S2O8-2
respectively.
You will determine the four absorptivities (one for each ion at each
wavelength). From the measured absorbance of the mixture at each wavelength, you
will have a system of two equations in two unknowns which can be algebraically
solved for the molar concentrations of the two ions.
A440 = A1 + A2 = Cr440 .b. [Cr] + Mn 440 .b. [Mn].. (1)

A545 = A1 + A2 = Cr545 .b. [Cr] + Mn 545 .b. [Mn] .. (2)
Where b is the thickness of the cuvette and i is the molar adsorption

coefficient at wavelength .
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The i constants can be determined experimentally by measuring the

absorbance of pure solutions of MnO 4- and Cr2O72-. A calibration curve is
prepared at each wavelength for both species. The slopes for concentration as a
function of Absorbance A = f(Ci), are used to calculate the K i values
Once this is done, the concentrations of Mn 7+ and Cr6+ can easily be
determined from the above two equations.
IV- Procedure:
Determination of Mn440 and Mn545
1- Add 1.00, 2.00, 3.00, and 4.00 ml of your 0.018 M standard MnO4-solution
in 25.0 ml standard flasks. Add in each 3.0 ml conc H 2SO4.
2- Add approximately 15 ml of distilled water, then add distilled water to the
mark, then mix well.
3- Measure the absorbance (A) between the wavelengths of 400 to 600 nm
(with 20 nm increments) using the most concentrated solution you have
prepared and plot A = f().
4- Prepare your blank solution by adding 3 ml of sulfuric acid to a 25 ml
standard flask and then adding distilled water to the mark. Verify that your
spectrum possesses two maxima, one at 525 and another at 545 nm.
5- Measure the absorbance A at 545 and 440 nm for all your prepared solutions
and plot: A = f (C) at the two different wavelengths on the same page.
Determination of Cr440 and Cr545

Repeat the same procedure as for Mn7+, but this time using a 0.015M standard
solution of Cr2O72-.
Analyzing a mixture of Cr(IV) and Mn(VII)

Measure the absorbance of the two unknown mixtures at 440 and 545nm.
Note that dichromate ion contains hexavalent chromium (Cr6+), which is a known
human carcinogen. We will collect all waste dichromate and permanganate in a
waste container in the hood. YourData
lab instructor
Tables needs to approve your safety
summary before you go on to Step 3 of this experiment.
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Solution MnO4- Cr2O72- Unknown Unknown
Flask Number 1 2 3 4 1 2 3 4 1 2
Volume of Standard 1.00 2.00 3.00 4.00 1.00 2.00 3.00 4.00
Concentration of Solution
Absorbance at (440nm)
Absorbance at (545nm)
Questions
1- From your plot of A= f(C) for MnO42- calculate the constants Mn440 and Mn545.
2- From your plot of A= f(C) for Cr2O72- calculate the constants Cr440 and Cr545.
3-From your plot of A= f(C) for MnO42- and Cr2O72 in each of the two unknown samples.
4-Why do we add concentrated sulfuric acid to the permanganate and dichromate solutions?
5- What is the significance of an absorption spectrum in identification of compounds?
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Exp 14
The Formula of a Complex Ion: Mole Ratio Method
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Exp 14
The Formula of a Complex Ion: Mole Ratio Method
As you have studied before, transition metals form coordinate covalent bonds
with Lewis bases to make coordination complexes. The anion or neutral compound
reacting with the metal are called ligands. A general reaction for the formation of
the complex follows:
One of the characteristics of transition metal complexes is that they absorb light
in the visible region of the spectrum. The instrument used for analysis is a
spectrophotometer. You know that the instrument works by passing a beam of light
through a cuvette, which contains the solution in a closed chamber. The beam
passes through the sample and into a detector. The detector measures the intensity
of the light that was passed through the solution, as a measurement of percent
transmittance. Photons are passed through the solution and the absorbing species
transitions to a higher energy state. Because the species is in and excited state and
unstable, the photons are then released and the species returns to the ground energy
state. Photons are released randomly from the absorbing species, which means as
the concentration of the absorbing species increases, the amount of photons
reaching the detector decreases. The equation to convert transmittance to
absorbance was previously discussed.
II- Purpose:
1- To practice spectrophotometric measurements.
2- To determine the stoichiometry of a metal complex by spectrophotometric means.
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II- Theory:
Perhaps the simplest of the spectrophotometric techniques that have been used
for the study of complex-formation equilibria is the molar ratio method. A series of
solutions are prepared. These solutions contain equal formal concentrations of a
metal ion but different formal concentrations of the ligand. The ratio of these
concentrations should usually vary from about 0.1 to 10 or 20. The absorbance of
each solution is then measured. If only the complex absorbs at the wavelength
where measurements are taken, then these absorbances are proportional to the
equilibrium concentrations of the complex ion in the solutions, and a plot of the
absorbance against the ratio of the number of moles of ligand to the number of
moles of metal ion (which is the same as the ratio of the corresponding total or
formal concentrations) will resemble This figure.
Typical data for a mole-ratio experiment.
The extent of the curvature in the vicinity of the end point depends on the
degree of dissociation of the complex. However, the stoichiometric formula of the
complex can be found by extrapolating the straight-line portions of the graph, which
is to say that the point at which these lines intersect corresponds directly to the ratio
of ligand to metal ion in the complex. This procedure works very well for weakly
dissociated (i.e. mostly associated) complexes.
But if the dissociation constant of the complex is too large, the molar ratio plot
will become a smooth continuous curve and it will be impossible to locate the
stoichiometric point. In such cases, better results can often be secured by the
continuous-variations methods. Within a certain rather restricted range, however,
the curvature around the "end point" of a molar ratio plot can be turned to good
advantage and used for the calculation of the dissociation constant of the complex.
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Continuous-Variation Method or Job`s Method:
This method is performed by preparing several solutions consisting of varying

amounts of the metal ion and complexing agent, however the sum of the metal ion
concentration and the ligand concentration is constant for each solution, The
absorbance of each solution is measured and plotted against the mole fraction of
metal ion or mole fraction of ligand.
The mole fraction of the metal, M, is defined as Cm
Cm +Cl
where Cm = the concentration of the metal and Cl = the concentration of the ligand.
A similar expression can be written for the mole fraction
(Cm +of
Clthe
) ligand.
From the graph, as the mole fraction of ligand

increases, there comes a point when the metal and
ligand exist in stoichiometric quantities.
Therefore, the absorbance reaches a maximum. As
more ligand is added and metal is taken away, the
absorbance decreases again.
Conditions that must be met in order for Job's method to be applicable .[
1. The system must conform to Beer's law

2. One complex must predominate under the conditions of the experiment
3. The Total concentration of the two binding partners must be maintained
constant
4. pH and ionic strength must be maintained constant
Mole ratio method:

In the mole ration method, varying amounts of ligand are added to a constant
amount of metal ion. The absorbance of each solution is measured and plotted
against the ration moles ligand/moles metal ions or against the variable volume of
the ligand in milliliters.This results a Beers law type calibration curve.
For a given complex stoichiometry, the graph will consist of a curve having two
straight line portions. Extrapolation of each straight line portion to an intersection
point gives the ratio of moles ligand/moles metal ion from the x-axis.
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Based on graphical technique, The plot shows you

the effect on absorbance when ligand is added to the
metal solutions. The absorbance increases until the metal
and ligand are present in stoichiometric quantities (after
which the absorbance levels off). Thus you can calculate
the amount of metal and ligand and determine the
formula or mole ratio of the complex.
In this experiment, you will determine the stoichiometry of the reaction between
Fe3+ and 5-sulfosalicylic acid; another daughter compound of salicylic acid. You
will prepare a series of solutions in which you will systematically vary the relative
amounts of Fe3+ and 5- sulfosalicylic acid. The intensity of the color of the
resulting mixtures will be used to identify the stoichiometric ratio in which Fe3+ and
5-sulfosalicylic acid react. The most intense color will be found in the solution in
which the two reactants have been combined in the correct ratio
for the reaction. Although you should be able to identify the most intensely color
solution with the eye, a spectrophotometer or colorimeter will be used to quantify
the color intensities. Solutions appear colored because components in the solution
absorb some wavelengths of visible light, leaving other wavelengths to be seen by
the eye. The colorimeter measures what percentage of light entering a solution is
able to pass through the solution without being absorbed. The equation can be
written as:
Fe3+ + nL Fe(L)n3+[
It is The absorbance increases until the metal and ligand are present in
stoichiometric quantities, after which the absorbance levels off. Thus you can
calculate the amount of metal and ligand, and determine the formula or mole ratio
of the complex.
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IV-Procedure:
A- Chemicals
1- Solution I: 0.0015M Ferric nitrate solution.
2- Solution II: 0.0015 M sulfosalicylic acid solution.
* Weigh 0.033 ( 10%) g of sulfosalycilic acid (MW=254) and transfer
to a 100.0 ml volumetric flask.
* Dissolve in 1 10-3 M HClO4 (Solution III), and dilute to the mark
with the same solution.
3- Solution III : 1 10-3 M HCIO4 or HCl Solutions.
B- Set Up
Set up your burette and fill it with solution III.
Make up nine solutions using test tubes with the proportions shown in the
table below.
Test tube label a b C d e f g h i

Solution I, ml 3.00 3.00 3.00 3.00 3.00 3.00 3.00 3.00 3.00
Solution II, ml 0.00 1.00 2.00 2.50 3.00 4.00 6.00 9.00 12.00
Solution III, ml 12.00 11.00 10.00 9.50 9.00 8.00 6.00 3.00 0.00
Mix each of the solutions thoroughly.

Measure their absorbance at a wavelength setting of 502 nm, using
solution III as a blank.
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Data Tables
Solution No. Solution % Tmeasured Ameasured

Composition
a 3.00 ml Fe3+
b 3.00 ml Fe3+
c 3.00 ml Fe3+
d 3.00 ml Fe3+
e 3.00 ml Fe3+
f 3.00 ml Fe3+
g 3.00 ml Fe3+
h 3.00 ml Fe3+
I 3.00 ml Fe3+
Questions
1- Compare briefly between the mole ratio method and the continous variation
method.
2- The following data were obtained for the Volume of Absorbance
-
spectrophotometric titration of 10.00ml of 3.8x10 Nitroso R, ml at 500 nm
5 2+ 0.00 0.000
M of Pb ions with Nitroso R ligand.
1.00 0.147
2+
Rxn:Pb + nL PbLn 2.00 0.271
3.00 0.375
a) Plot the titration curve (Absorbance verses 4.00 0.371
5.00 0.347
volume of ligand).
6.00 0.325
b) Indicate the volume of ligand at equivalence 7.00 0.306
point. 8.00 0.289
c) If the concentration of the ligand is 2.44x10-
4
M, calculate the coordination number n of lead Pb2+ in the above reaction.
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Experiment 15
Determination of the Equilibrium Constant for a Chemical Reaction
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Experiment 15
Determination of the Equilibrium Constant for a Chemical Reaction
I- Purpose:
1- To review the concepts and principles of chemical equilibrium.
2- To learn the principles and use of a spectrophotometer.
3- To learn the principles of chemical analysis by spectrophotometric means.
4- To determine, spectrophotometrically, the equilibrium constant for the
reaction between iron (III) ion and thiocyanate ion (SCN -) at a given
temperature.
II- Theory:
When chemical substances react, the reaction typically does not go to
completion. Rather, the system goes to some intermediate state in which both the
reactants and the products have concentrations which do not change with time:
Chemical Equilibrium is reached.
When a reaction mixture reaches equilibrium at a particular temperature, it
obeys the Law of Chemical Equilibrium which imposes a condition on the
concentrations of reactants and products. This condition is expressed in the
Equilibrium Constant K, for the reaction at a given temperature.
In this experiment, we study the equilibrium reaction:
Fe3+(aq) + SCN-(aq) FeSCN2+(aq)
The equilibrium constant is defined as:
[FeSCN 2 ]
Kc
[Fe3 ][SCN ]
Kc is constant at constant temperature; this means that mixtures containing

Fe and SCN- will react until this equilibrium is satisfied, so that the same value of
3+
Kc will be obtained no matter what initial amounts of Fe3+ and SCN- are used.
This reaction is particularly suitable for study because Kc has a convenient
magnitude and the color of the FeSCN2+ ion allows for easy spectrophotometric
analysis of the equilibrium mixture. FeSCN2+ is a deep, blood-red complex with an
absorption maximum at 447 mm. Kc will be determined for several mixtures made up
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in different ways, and it can thus be shown that Kc has indeed the same value in each
mixture.
The Beer-Lambert's Law will be applied, in this experiment, to determine the
FeSCN2+ concentration at equilibrium. A = bc, where :
2- A is the absorbance of the substance
3- is the molar absorptivity coefficient (it is constant at a given wavelength
for a particular absorbing substance)
4- b is the width of the light path that passes through the absorbing substance
in centimeters
5- c is the molar concentration of the absorbing species.
In the first part of the experiment, a set of standard solutions of the FeSCN2+
complex is prepared using a concentration of Fe3+ that far exceeds the SCN-
concentration. This huge excess of Fe3+ pushes the equilibrium far to the right, nearly
consuming all the original SCN- concentration. The reaction is thus assumed to go to
completion. The absorbance of each solution is measured, then plotted versus the
molar concentration of FeSCN2+; this establishes a calibration curve from which the
concentrations of FeSCN2+ are determined for the chemical systems in the second
part.
In the second part of the experiment, you will prepare a set of various solutions
using nearly the same concentration of Fe3+ and SCN- ions, and thus have a number
of equilibrium systems. By knowing the initial concentrations of Fe3+ and SCN-, and
by determining the equilibrium concentration of FeSCN2+ spectrophotometrically, the
equilibrium concentrations of Fe3+ and SCN- can be calculated. Using these
equilibrium concentrations, the Kc for the system is calculated.
Calculation:
Fe3+(aq) + SCN-(aq) FeSCN2+(aq)
Initially CoFe3+ CoSCN- 0
Change -x -x x But x = CFeSCN2+
At equil. C0 Fe3+ - CFeScN2+ C0 SCN- - CFeSCN2+ CFeSCN2+
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Where
CFeSCN2+ = x = [FeSCN2+] = (A/b)
A is the measured absorbance for that particular solution.
Note: In preparing the mixtures for this experiment, the H+ ion concentration
will be maintained at 0.5 M; H+ does not participate directly in the reaction, but its
presence is necessary to avoid the formation of brown color species such as
Fe(OH)2+, which interferes with the analysis of FeSCN2+.
III- Procedure:
Solutions: Solution A: 2.00 10-3 M Fe(N03)3 in 1.0 M HN03

Solution B: 2.00 10-3 M KSCN
Solution C: 0.200 M Fe(NO3)3 in 1.0 M HNO3
Place 30 ml of solution A in a clean 100 ml beaker. Then in another clean

100 ml, beaker, place 20 ml of solution B. Finally, in a third beaker, put
40 ml of solution C.
Part I:
1- Obtain from the storeroom 12 clean test tubes. Label them from 0 to l 1.
2- Pipet 5.00 ml, of solution C into each of the tubes 0 to 5.
3- To the tubes 0 to 5, add 0.00, 0.50, 1.00, 1.50, 2.00 and 3.00 ml of solution
B respectively. Then add distilled water to each of the tubes so that the
final volume is 20.00 ml in each of them. Deliver the necessary volume of
water by means of a buret.
4- Cover the tubes with parafilm paper and mix the solutions well to make
them homogeneous.
5- Measure the absorbance of each of the prepared solutions (using a
spectrophotometer) at a wavelength = 447 nm. Record your data in Table
1.
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Part II:
6- Pipet 5.00 ml of solution A into each of test tubes 6 to 11.
7- To the test tubes 6 to 11, add 0.00, 1.00, 2.00, 3.00, 4.00 and 5.00 ml of
solution B respectively. Then add distilled water to each of the tubes so that
the final volume is 10.00 ml in each of them. Deliver the necessary volume
of water by means of a buret.
8- Cover the tubes with parafilm paper and mix the solutions well to make
them homogeneous.
9- Determine the absorbance of each of the five prepared solution, at a
wavelength 447 nm. Record your data in Table 2.
Data Tables
Table 1: Standard Solutions and Absorbance Measurements @ = 447 nm for the Calibration Curve
V V V
Mixture Absorbance [FeSCN2+]
(Fe(NO3)3) (KSCN) (H2O)
0 5.00 0.00 15.00
1 5.00 0.50
2 5.00 1.00
3 5.00 1.50
4 5.00 2.00
5 5.00 3.00
Slope of Calibration Curve: .b(M-1) = ___________________
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Table 2: Composition of Mixtures and Absorbance Measurement @ = 447 nm
V V V
Mixture Absorbance [FeSCN2+]
(Fe(NO3)3) (KSCN) (H2O)
6 5.00 0.00 5.00
7 5.00 1.00
8 5.00 2.00
9 5.00 3.00
10 5.00 4.00
11 5.00 5.00 0.00
Table 3: Equilibrium Concentrations and Equilibrium Constant

Mixture C0Fe3+ C0SCN- [Fe3+] [SCN-] [FeSCN2+] Kc
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Questions
1- Show one sample calculation for each calculated quantity (for one mixture
only).
2- Consider the chemical reaction:

HClO2 (aq) H+ (aq) + ClO2- (aq)
What are the equilibrium concentration of H+, ClO2- and HClO2 in an
aqueous solution with initial concentration [HClO2]0 = 0.260 M, given that
the reaction has an equilibrium constant Kc = 1.1 10-2?
3- Back to the chemical reaction studied in this experiment:

Fe3+ (aq) + SCN- (aq) FeSCN2+ (aq)
We mix 2.00 ml of a 1.00 10-3 M Fe(NO3)3 with 3.00 ml of a 1.75 10-3
M KSCN solution. What are the equilibrium concentrations of Fe3+, SCN-
and FeSCN2+? (Use the value Kc= 1.15 102)
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Experiment 16
Potentiometric Titration
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Experiment 16
Potentiometric Titration
Many Acid-Base titrations are difficult to accomplish using a visual indicator

for one of several reasons. Perhaps the analyst is color-blind to a particular indicator
color change; there may not be a suitable color change available for a particular
type of titration or the solutions themselves may be colored, opaque or turbid. It
may be desired to automate a series of replicate determinations. In such situations,
potentiometric titration, using a glass hydronium ion selective electrode, a suitable
reference electrode and a sensitive potentiometer (a pH meter) may be
advantageous.
Potentiometry is one type of
electrochemical analysis methods.
Electrochemistry is a part of chemistry,
which determines electrochemical
properties of substances. An electrical
circle is required for measuring current and
potential (voltage) created by movement of
charged particles. Galvanic cell serves as a
sample of such system.
Electrochemical cell consists of two solutions connected by salt bridge and
electrodes to form electrical circle. Sample cell on the above figure consists of
solutions of ZnSO4 and CuSO4. Metallic Zn and Cu electrodes are immersed in
respective solutions.
Electrodes have two contacts:
a) through wires connected tovoltmeter;
b) through solutions and salt bridge.
Salt bridge consists of a tube filled with saturated salt solution (e.g. KCl
solution). The ends of the tube are capped with porous frits that prevent solutions
from mixing, but permit movement of ions.
Three distinct charge transfer processes are described for the system:
1. Electrons move in electrodes and wires from zinc electrode to copper electrode.
2. Ions move in solutions:

a. In solution on the left, zinc ions move away from the electrode and sulfate ions
move towards it.
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b. In solution on the right, copper ions move towards the electrode and negatively
charged
ions (sulfate) away from it.
c. In salt bridge positive ions move right and negative ions left.
3. On the surfaces of electrodes electrons are transferred to ions or vice versa:

a. Zinc electrode dissolves.
b. Metallic copper is deposited on the electrode surface.
Three processes mentioned above form a closed electrical circle making the
flow of electrical current possible.
Potential on an electrode depends on the ions present in the solution and their
concentration. This way electrochemical cells can be used to determine ions and
their concentration in solution. The dependence of potential between electrodes
from concentration of ions is expressed by Nernst equation discussed before.
II- Purpose:
1- To measure the change in the potential of Fe2+ solution by the titration of the
ferrous solution with K2Cr2O7 solution.
2- To plot the value of Emeasured versus ml K2Cr2O7 added and form the plot to
determine the equivalent point and mid-point of the titration.
3- To calculate Eo of Fe3+/Fe2+ from the measurement of the mid-point
titration (E1/2).
III- Theory:
Potentiometry is an electroanalytical
method which is based on measurement of
potential of an electrode system. Potentiometric
measurements enable selective detection of
ions in presence of multitude of other
substances.
Potentiometric measurement system
consists of two electrodes, potentiometer and a
solution of analyte. In system like one depicted
on the next figure, the potential is measured in
reference to calomel electrode e.g. calomel
electrode functions as reference electrode.
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Reference electrode is an electrode with potential which is

a) independent of analyte (or other) ions in solution;
b) independent of temperature.
Most commonly, the reference electrode is the silver/silver chloride electrode.
Ag(s) + Cl-(aq)
AgCl(s) + e
The electrode sensitive to hydrogen ions is an indicator electrode. Potential of

an indicator electrode depends mainly on the concentration of the analyte ions (in
this case hydrogen ions).
The Mechanism of the Response

A change in hydronium ion concentration causes a change in composition of
the glass membrane due to an ion exchange process involving the solution and the
membrane (see textbook for details). A corresponding change in membrane
potential, proportional to pH, is what is measured.
All other potentials are constant. In effect the membrane potential (variable) is
measured against two fixed potentials, the external reference and the internal
reference, both Ag/AgCl reference electrodes.
Potential difference is measured using a high

impedance potentiometer. This high resistance 12.00
dictates a very small current flow.

Any acid-base titration may be conducted 10.00
potentiometrically. Two electrodes, after

8.00
calibration [to relate potential in millivolts (mV) to
a pH value] are immersed in a solution of the 6.00
analyte. One is an indicator electrode, and the

4.00
other a stable reference electrode. The potential
difference, which after calibration is pH, is 2.00
measured after the successive addition of known 0.00 5.00 10.00 15.00 20.00 25.00
increments of acid or base titrant.
When a potentiometric titration is being performed, interest is focused upon

changes in the emf of an electrolytic cell as a titrant of known concentration is
added to a solution of unknown. The method can be applied to all titrimetric
reactions provided that the concentration of at least one of the substances involved
can be followed by means of a suitable indicator electrode.
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The critical problem in a titration is to recognize the point at which the

quantities of reacting species are present in equivalent amounts. The titration curve
can be followed point by point, plotting as ordinant, successive values of the cell
emf (pH) vs the corresponding volume of titrant added. A typical titration curve is
presented here. another method for determining the equivalence point from the
titration curve data is shown here also.
The potential that is measured in this experiment is that between ferrous-ferric

couple and the saturated calomel electrode:
Emeasured = EFe3+Fe2+ - Ecalomel
For the half reaction: Fe3+ + e- Fe2+, the Nernst equation is:
0.059 [Fe2 ]
E Fe3 / Fe2 E O
Fe3 / Fe2
log
1 [Fe2 ]
For the calomel: Ecalomel = 246 mV
0.059 [Fe2 ]
then E measured E O
Fe3 / Fe2
log 246mv
1 [Fe2 ]
When K2Cr2O7 solution is added to a ferrous solution, the following reaction

takes place:
6Fe2+ + Cr2O72- + 14H+ 6Fe3+ + 2Cr3+ + 7H2O
Accordingly, if a ferrous solution is titrated by Cr2O72- of known
concentration (K2Cr2O7 can be a primary standard), the potential of EFe3+/Fe2+
changes according to [Fe2+] and [Fe3+] in the solution.
An important point in the titration is the mid-point titration where:
amount of Fe3+ = amount of Fe2+
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or [Fe2+] = [Fe3+],
then log ([Fe2+]/[Fe3+]) = log 1 = 0
Emeasured (called E1/2) = EoFe3+Fe2+ - 246 mv Eo Fe3+Fe2+ = E1/2 + 246 mv
Thus if a titration curve is plotted for Emeasured versus ml of Cr2O72- added, and
the equivalence point determined from the plot, then the Emeasured at mid-point in
titration can be determined (where if the ferrous solution and K2Cr2O7 are of same
molarities at 1/2 V, and V is the volume of K2Cr2O7 needed to reach the endpoint),
consequently Eo Fe3+Fe2 value can be calculated.
IV- Procedure:
1. Preparation of the standard dichromate solution:
- Weigh accurately by difference about 0.3 grams of K2Cr2O7.
- Record the exact weight of the K2Cr2O7.
- Transfer the K2Cr2O7 to a 100.0 ml volumetric flask.
- Add some distilled water to the flask and swirl till K2Cr2O7 dissolves.
-When the entire sample is dissolved, add water slowly to the mark.
Cover and homogenize.
2. Potentiometric titration of a ferrous salt with the standard dichromate:

- Obtain a sample of ferrous salt with an appropriate acid solution. If the
salt is FeC12, the acid should be HCl, or if the salt is FeSO4, the acid
should be H2SO4. Weigh exactly about 1.000 millimol of the solid into a
250 ml beaker. Then add 90 ml of 1M acid and 90 ml of distilled water,
stir to dissolve.
- Titrate immediately with the standard dichromate solution, adding 1.00 ml at a
time, measuring the electric potential after each addition of titrant. In the vicinity
of the end point, use smaller increments of titrant. You may need to wait for
about 1/2 a minute after each addition for a stable reading to be reached.
- After the end-point is reached add 1.00 ml increment and take an additional five
to six readings
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Data Tables
Weight of K2Cr2O7 g
Weight ferrous salt g
Volume of Volume of Volume of

K2Cr2O7 E(mV) K2Cr2O7 E(mV) K2Cr2O7 E(mV)
added added added
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Questions
1- Draw on a graph paper the plot of Electric potential along the ordinate Vs
ml of dichromate solution along the abscissa.
2- Show on the graph the volume of titrant needed to reach endpoint, the
corresponding E value and the E1/2 Value.
3- Calculate the Eo for the half reaction Fe3+ + e- Fe2+.
4- Calculate the theoretical titrant volume.
5- Compare the experimental titrant volume with the theoretical volume,
comment on the different in their values and find the % error.
6- Comment on the % error, and then discuss all possible sources of error in
the experiments.
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Experiment 17
Analysis of Analgesic Tablets by High Performance Liquid Chromatography
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Experiment 17
Analysis of Analgesic Tablets by High Performance Liquid Chromatography
I- Introduction:
Analgesics are prepared in various formulations that often include one or
more of thefollowing compounds: acetylsalicylic acid (aspirin, hereafter
abbreviated SA), acetaminophen (the active ingredient in Tylenol, hereafter
abbreviated AC), and caffeine (a vasodilator that accelerates the delivery of the
active ingredients, hereafter abbreviated CAF).
Certain over-the-counter (OTC) pain relievers contain all three of these
ingredients, and this combination is marketed as a particularly effective remedy for
headaches. Pharmaceutical manufacturers must maintain tight quality control of
their products, and routinely use chromatographic methods for the analysis of
medications.
II- Purpose:
In this lab you will perform a quantitative determination of the CAF and SA content
of an over-the-counter headache tablet using HPLC.
III- Theory
According to the label on the bottle, each tablet has the following content:
Acetaminophen: 250 mg
Acetylsalicylic acid: 250 mg
Caffeine 65 mg
Filler 100 mg
Caffeine
Filler is an inert material that should be

water soluble. It is of no interest to us in this analysis,
though pharmaceutical manufacturers
must also control its content.
Acetaminophen
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You will measure the concentrations of CAF and SA in a single tablet by

crushing the tablet and extracting the active ingredients in preparation for analysis
by HPLC. You will also prepare a set of 4 standards containing both CAF and SA
in order to make a quantitative determination of the mass of these compounds in the
tablet.
Chromatographic method:
We will use an isocratic (constant solvent mixture) method for this determination.
The compounds of interest will be separated on a around 15 cm C18 column.
The mobile phase has been prepared for you. It is a four component mixture:
- 94.1 %Water (primary component of polar mobile phase)
- 5.5 %Acetonitrile (stabilizes the nonpolar stationary phase)
- 0.2 %Triethylamine (neutralizes polar sites on column)
- 0.2 %Acetic Acid (maintains carboxylic acids in the neutral, protonated form)
The mobile phase flow rate should be set to 1ml/min. You will monitor
absorbance at 4 wavelengths (203, 254, 263 and 280 nm). SA, CAF and AC absorb
strongly in this region of the spectrum.
IV-Procedure:
Sampling the lot of tablets:

When performing analyses for quality control, your first task is to devise an
appropriate method to prepare a representative sample of tablets. Normally this
would be done by selecting several tablets and analyzing them to determine the
average mass of the analytes of interest in each tablet and the variance of the mass
in the lot. We will only perform the analysis on a single tablet in the interest of
time.
Sample preparation:
1. Weigh several headache tablets and compute the average mass and the standard
deviation of the tablets.
2. Select one tablet for analysis and record its mass. Use a mortar and pestle to
crush the tablet into a fine powder.
3. Weigh about 20 mg of this powder directly into a 100 ml beaker. Record the
added mass. Add about 50 ml of mobile phase to the beaker to dissolve the powder.
Be careful not to use more than 50 ml because you will deliver this solution to a
100.0 ml volumetric flask for final dilution. The mobile phase is used to dissolve
the tablet so that the solvent does not alter the composition of the mobile phase in
the column.
4. Carefully pour the solution from the beaker to a clean 100.0 ml volumetric flask.
Rinse the beaker with about 5 ml of mobile phase 2 or 3 times, and pour these into
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the flask to make a quantitative transfer of the dissolved material to the volumetric
flask. Be sure to mix the solution by agitating it before you have filled the flask.
Pour a small volume of mobile phase into the beaker and use it to dilute the solution
in the flask to the mark. Mix the solution again by inverting the stoppered flask, and
label the flask SAMPLE. Label a screw top sample vial and transfer about10 ml
of the sample to the vial.
If the actual contents of the tablet are equal to the content stated on the label, the
sample will contain CAF and SA in the following concentrations:
- CAF: 1.955 mg in 100 ml solution (0.01955 mg/ml)
- SA: 7.519 mg in 100 ml solution (0.07519 mg/ml)
We use mg/ml as our unit of concentration because we will ultimately want to
calculate the mass of each analyte in the tablet.
Standard Set UP:

Next you will prepare standards that contain both CAF and SA in the approximate
concentrations listed below:
Standard label Conc. of SA Conc. of CAF

1 0.08000 mg/ml 0.0150 mg/ml
2 0.07500 mg/ml 0.0200 mg/ml
3 0.07000 mg/ml 0.0250 mg/ml
4 0.06500 mg/ml 0.0300 mg/ml
You do not need to use these exact concentrations, but you need to record precise
mass values so that the actual concentrations are accurately known. You will notice
two features of this set of standards.
First the concentrations vary with opposing trends for each analyte. This allows
you to make an unambiguous qualitative determination of the retention time of each
analyte, since one will have an increasing peak height as the standard number
increases, and the other will have a decreasing peak height.
Secondly the standard concentrations closely bracket the expected
concentrations of the analytes. This is appropriate because we expect a very narrow
variation in the concentration range of the sample.
Stock Solutions:
Prepare separate stock solutions of SA and CAF as follows:
1- SA: Label a 25.0 ml volumetric flask SA STOCK. Weigh about 16 mg of
SA and deliver it into a 25.0ml volumetric flask. Note the actual mass
delivered to the flask in your notebook. Add about 12 ml of mobile phase and
dissolve the SA. Dilute to the mark to give a stock solution containing about
0.64 mg/ml. Using the actual mass, calculate the exact concentration of the SA
stock solution and note it in your notebook.
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2- CAF: Label a 25.0 ml volumetric flask CAF STOCK. Weigh about 6 mg of

CAF and deliver it into a 25.0 ml volumetric flask. Note the actual mass
delivered to the flask in your notebook. Add about 12 ml of mobile phase and
dissolve the CAF. Dilute to the mark to give a stock solution containing
About 0.24 mg/ml. Using the actual mass, calculate the exact concentration
of the CAF stock solution and note it in your notebook.
Standards Preparation
Label four 10.0 ml volumetric flasks and add the following volumes of the stock
solutions to each flask with a 2.00 ml graduated pipette:
Standard label volume of SA stock soln. volume of CAF stock soln
1 1.250 ml 0.625 ml
2 1.172 ml 0.833 ml
3 1.094 ml 1.042 ml
4 1.016 ml 1.250 ml
Dilute each flask to the mark with mobile phase. Label 4 screw top sample vials,
transfer the standards to the vials and seal them.
Measurement and Analysis:
- Collect a chromatogram of each standard and the sample by injecting 20 ml of

each solution onto the HPLC using the chromatographic method outlined above.
The standards should give two peaks (SA and CAF) and the sample should give
three (SA, CAF, and AC).
- Make a table of the results for the standards using the 263 nm chromatogram
including the peak area, width and retention time of each peak at each
concentration. Make a table of the same parameters for each peak of the sample
chromatogram.
- Use the peak areas of the 263 nm chromatogram and known concentrations of
the standards to make calibration plots of concentration vs. peak area. Are these
plots linear? They should be!
- Perform a linear regression to determine the slope and y-intercept of the
equations.
AreaSA = mSA CSA + bSA
AreaCAF = mCAF CCAF + bCAF
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- Note these values in your notebook. Be sure to include the standard errors for the
slopes and intercepts. Use these values and the peak areas from your unknowns
to determine the concentrations of each component in the sample solution. Use
the known sample dilution, the actual mass of sample analyzed and the known
tablet mass to calculate the mass of CAF and SA in the pain reliever tablet that
you analyzed.
- A formal report format will be used for this lab. While the reports that each
student turns in should be typed, ALL OBSERVATIONS SHOULD BE RECORDED
IN YOUR LAB NOTEBOOKS.
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Experiment 18
Determination of Chloride Ions by Volhard`s Method
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Experiment 18
Determination of Chloride Ions by Volhard`s Method
I- Introduction:
Precipitation titrimetry, which is based on reactions that yield ionic

compounds of limited solubility, is one of the oldest analytical techniques. The
slow rate of formation of most precipitates, however, limits the number of
precipitating agents that can be used in titrations to a handful. The most widely
used and important precipitating reagent, silver nitrate, is used for the
determination of the halogens, and the halogen-like anions. Titrations with
silver nitrate are sometimes called argentometric titrations.
Titrations in which precipitates are formed are called precipitation titrations.

The most frequent application of this type of titration uses silver ion to
determine chloride. According to the indicator used, three methods can be
described. Chromate is the indicator in Mohr's method while Fajans method
makes use of adsorption indicators. Both methods are direct methods. The third
method is an indirect method where an excess silver is added to chloride
unknown and the remaining silver is back-titrated with a standard thiocyanate
solution in presence of Fe(III) as an indicator.
Titration curves for precipitation reactions are derived in a completely

analogous way to the methods described for titrations involving strong acids and
strong bases. P-functions are derived for the preequivalence-point region, the
postequivalence point region, and the equivalence point for a typical
precipitation titraton.
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Most indicators for argentometric titrations respond to changes in the

concentration of silver ions. As a consequence, titraton curves for precipitation
reactions usually consist of a plot of pAg versus volume of AgNO3.
II- Theory of The Volhard Method (Colored Complex)

This method uses a back titration with potassium thiocyanate to determine the
concentration of chloride ions in a solution. Before the titration an excess volume of
a silver nitrate solution is added to the solution containing chloride ions, forming a
precipitate of silver chloride. The term excess is used as the moles of silver nitrate
added are known to exceed the moles of sodium chloride present in the sample so
that all chloride ions present will react.
Ag+ (aq) + Cl (aq) AgCl(s)
The indicator Fe3+ is then added and the solution is titrated with the potassium
thiocyanate solution. The titrate remains pale yellow as the excess (unreacted) silver
ions react with the thiocyanate ions to form a silver thiocyanate precipitate.
Ag+(aq) + SCN(aq) AgSCN(s)
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Once all the silver ions have reacted, the slightest excess of thiocyanate reacts with
the indicator Fe3+ to form a dark red complex. The solution turns red with the first
slight excess of thiocyanate ion:
Fe3+(aq) + SCN(aq) [FeSCN]2+(aq)
The concentration of chloride ions is determined by subtracting the titration

findings of the moles of silver ions that reacted with the thiocyanate from the total
moles of silver nitrate added to the solution. This method is used when the pH of
the solution, after the sample has been prepared, is acidic. This is to prevent
precipitation of iron (III) as the hydrated oxide or Fe(OH)3. If the pH is neutral or
basic, Mohrs method or the gravimetric method should be used. The method is
illustrated below by using the procedure to determine the concentration of chloride
(from sodium chloride) in cheese.
Note:
The use of acidic medium together with added SCN- titrant increase the
solubility of the precipitate leading to significant errors. This problem had been
overcome by two main procedures:
1. The first includes addition of some nitrobenzene, which surrounds the
precipitate and shields it from the aqueous medium.
2. The second procedure involves filtration of the precipitate directly after
precipitation, which protects the precipitate from coming in contact with the
added SCN- solution.
Theory of Mohr Method
This method utilizes chromate as an indicator. Chromate forms a precipitate

with Ag+ but this precipitate has a greater solubility than that of AgCl, for
example. Therefore, AgCl is formed first and after all Cl - is consumed, the first
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drop of Ag+ in excess will react with the chromate indicator giving a reddish
precipitate.
2 Ag+ + CrO42- = Ag2CrO4
In this method, neutral medium (about 7 ) should be used since, in alkaline
solutions, silver will react with the hydroxide ions forming AgOH. In acidic
solutions, chromate will be converted to dichromate.
Note:
There is always some error in this method because a dilute chromate solution is
used due to the intense color of the indicator. This will require additional
amount of Ag+ for the Ag2 CrO4 to form.
Theory of Fajans Method
Fluorescein and its derivatives are adsorbed to the surface of colloidal AgCl.
After all chloride is used, the first drop of Ag + will react with fluorescein (FI-)
forming a reddish color.
Ag+ + FI-= AgF
Since fluorescein and its derivatives are weak acids, the pH of the solution
should be slightly alkaline to keep the indicator in the anion form but, at the
same time, is not alkaline enough to convert Ag + into AgOH . Fluorescein
derivatives that are stronger acids than fluorescien (like eosin) can be used at
acidic pH without problems. This method is simple and results obtained are
reproducible.
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III- Reagents and Chemicals
1. Chloride solution of unknown concentration.

2. Pre-dried and desiccated AgNO3 . Be careful !!
Do not allow this substance either in the solid or solution to come in
contact with your skin. Black stains that will persist for almost two
weeks will be formed.
3. 6 M HNO3 solution.
4. Predried and desiccated KSCN: 0.01 M solution : Prepare by dissolving
a suitable amount (2.43g) in 250 ml of distilled water and standardize
against standard AgNO3 solution using ferric alum indicator.
5. Ferric alum indicator: Saturated Ferric ammonium sulfate solution: Add
8g of NH4Fe(SO4)2.12H2O to 20 ml of distilled water and add a few drops of
concentrated nitric acid.
6. Standard AgNO3 : Silver nitrate solution: (0.1 M). If possible, dry 5 g of
AgNO3 for 2 hours at 100C and allow to cool. Accurately weigh about 4.25
g of solid AgNO3 and dissolve it in 250 ml of distilled water in a conical
flask. Store the solution in a brown bottle.
7. Potassium permanganate solution: (5%) Add 1.5 g KMnO4 to 30 ml of
distilled water.
IV- Procedure: Determination of Chloride by Precipitaiton Titration Using

Volhard Method
You can use a sample of unknown chloride solution or you can work on
cheddar cheese.
151
152 | P a g e
For unknown Chloride Solution:
1. Transfer exactly 5.0 ml of the chloride sample into a stoppered conical

flask.
2. Add 10 ml of distilled water, 2 ml of the 6 M HNO3 solution provided,
and exactly 10.0 ml of standard AgNO3 solution. You may add 5 ml of
nitrobenzene and shake vigorously, but you may need to seal the flask
with parafilm. This step can be skipped.
3. Add 1 ml of the ferric alum indicator and titrate against standard
KSCN solution and record the volume to two significant figures after
the decimal point.
4. Repeat steps 1-3 two more times and calculate the concentration of
chloride as ppm NaCl.
For Chloride in Cheddar Cheese:
The salt sodium chloride is added during the manufacture of cheddar cheese. In
this method, the cheese is digested to release this salt to obtain the concentration
of chloride ions. To carry out this digestion, the cheese is reacted with nitric acid
and potassium permanganate. The chloride ions are then free to form a precipitate
with the added silver ions.
1. Cut or grate the cheese into fine pieces and accurately weigh about 6 g into a 500
ml conical flask.
2. Precisely add 50.0 ml of 0.1M silver nitrate solution (by pipette if possible), 20
ml of concentrated nitric acid, (very carefully,check safety notes), 100 ml of
distilled water and a few boiling chips, and heat the solution to boiling in a
fumehood.
3. As the solution boils add 5 ml of 5% potassium permanganate solution. This

addition will cause a very smelly reaction so done in the fumehood. Keep boiling
until the purple colour disappears, then add another 5 ml of potassium
152
153 | P a g e
permanganate solution. Continue this process until 30 ml of potassium

permanganate solution has been added and the cheese particles are completely
digested (or as close as possible). To find out when digestion is complete, remove
the flask from heat and allow it to stand for a few moments. Undigested cheese
particles will float upon the surface of the clear liquid, while the white precipitate of
silver chloride will sink to the bottom. If there is still too much undigested cheese,
the boiling and addition of 5 ml of potassium permanganate should be continued,
checking each time until there is a satisfactory level of digestion.
4. Cool the solution and filter it. Wash the solid residue with a few ml of distilled
water.
5. Make the filtrate up to 500 ml in a volumetric flask.
Titration
1. Use a volumetric cylinder to measure 100.0 ml of the cheese extract solution
solution and pour it into a conical flask.
2. Add 1.0 ml of saturated ferric alum indicator.

3. Titrate the unreacted silver ions with the 0.1 M KSCN . The first appearance of
a dark red color is due to the ferric thiocyanate complex.
4. Repeat the titration with 100 ml samples of the cheese extract solution until you
obtain concordant results.
153
154 | P a g e
V- Results:
Vol of AgNO3 Vol of Vol ppm

KSCN AgNO3 NaCl
Eq to
Chloride
Mean
Questions
1. Determine the average volume of potassium thiocyanate used from your

concordant titres.
2. Calculate the moles of potassium thiocyanate used.
3. Use the equation of the reaction between silver ions and thiocyanate ions
Ag+(aq) + SCN(aq) AgSCN(s)
to calculate the moles of unreacted silver nitrate in your sample. In case of cheese
sample, multiply the figure by five to determine the total moles of unreacted
silver nitrate (the excess) in the 500 ml volumetric flask.
4. Calculate the moles of silver nitrate in the 50 ml of solution that was added
during the sample preparation to the cheese.
154
155 | P a g e
5. Calculate the total moles of silver nitrate that reacted with the salt from the
cheese by subtracting the moles of unreacted silver nitrate (the excess) from the
total moles of silver nitrate added to the cheese.
6. Use the equation of the reaction between the silver ions and the chloride ions to
calculate the moles of sodium chloride in the sample of cheese.
Ag+(aq) + Cl(aq) AgCl(s)
7. Calculate the concentration of sodium chloride in the cheese as grams of salt per
100 g cheese (% salt).
Additional Notes
1. Residues containing silver ions and precipitate are usually saved for later
recovery of silver metal. Check this with your teacher or the laboratory
supervisor.
2. A blank titration substituting sucrose (sugar) for the cheese should be carried
out to see if there are any contaminating chloride ions present in the reagent
solutions used. If any are found, the figure should be subtracted from the titration
results.
3. For greatest accuracy it is a good idea to standardise your thiocyanate solution by

titrating several samples against your standardised silver nitrate solution (once
again using ferric ammonium sulfate indicator). The concentration of SCN
determined by this titration should then be used in all calculations
155
156 | P a g e
Exp No. Title Page
- Laboratory Items 2
- Safety Regulations and Policy 8
- Notebooks & Lab Reports 11
Measurements and Errors 15
Part I Applications of Acid Base Titration 25
1 Preparation of Primary and Secondary Standards 26
2 Determination of the solubility of oxalic acid at room temperature 33
3 Determination of Acidity of Vinegar 34
4 Titration of Stomach Antacid 37
5 pH measurements and Buffer Capacity 41
6 Titration of a Polyprotic Acid: Phosphoric Acid 56
Part II EDTA Titrations 63
7 Total Water Hardness 64
Part III Redox Titrations 71
8 Determination Of Vitamin C in a Tablet 75
9 The Strength of Laundry Bleach 81
10 Determination of Calcium in Milk by Oxalate 88
11 Electrochemistry and Nernst Equation 94
Part IV Spectrophotometric Instrumental Analysis 102
12 Spectrophotometric Analysis of a commercial aspirin tablets 109
13 Quantitative Analysis for a Mixture of Two Ions 113
14 Determination of the Formula of a Complex Ion 118
15 Determination of the Equilibrium Constant 125
16 Potentiometric Titration 132
17 Analysis of Analgesic Tablets by HPLC 140
18 Determination of Chloride by Volhard Method 146
- Index 156
156

Quantitative Analysis: LAB MANUAL (Phar205)

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Quantitative Analysis: LAB MANUAL (Phar205)

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Lebanese International University

Absorbance versus volume of Ligand

0.0 Dr. Mohammad Ahmad Assi

volume of Nitros R (mL )

-1- LABORATORY ITEMS

Name Description Picture

Used to hold and heat liquids.

The Erlenmeyer Flask is used to heat

The Volumetric flask is used to measure

A graduated cylinder is used to

The dropper is used for moving small

Graduated pipettes, also called Mohr

The buret is used in titrations to

The Micro-spatula, commonly called a

The stir rods are used to stir things.

The funnel can be used to target liquids

The Mortar and Pestle are used to

Bottles can be used for storage, for

A wash bottle is a squeeze bottle with a

A low density polyethylene (LDPE)

A cuvette is a small tube of circular or

Crucibles are used to heat small

Evaporating The Evaporating Dish is used to heat

The triangle is used to hold crucibles

Tongs are used to hold many different

The clamp stand is used to hold

Buret Clamp A buret clamp is used to fasten

Ring Support A support ring

Test tubes are used to hold, mix, or

Test tube The holder is used to hold test tubes

The thermometer is used to take

Stoppers come in many different sizes.

Parafilm can be used to seal cuvettes,

Paper Towels are essential to the lab

Test tube The test tube brush is used to easily

-2- SAFETY REGULATIONS & Policy

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

-3- Laboratory Reports and Note Books

Students preparation for the experiment is usually performed in two weeks:

The lab notebook must:

1. A laboratory report will be required for each experiment. All experiments

4. The report should be complete,

5. Reports should be neat and attractive in appearance.

III. Purpose or Objective

IV. Equipments, tools and Chemical Reagents

VIII. Calculations and Results

Quantitative Classical Chemical Analysis

Gravimetry Volumetric Titrations Instrumental analysis

Acid-base Precipitation Complexometric Redox

Titrations involving iodine (I2)

Iodometric titration of copper

Titration Analyte Titrant Indicator

Acid-base Quantification Acetic acid NaOH (sodium Phenolphthalein

Complexometric Water Calcium and EDTA Eriochrome black

Redox Quantification Hydrogen peroxide KMnO4 No indicator

1. To learn how uncertainty in a measurement arises

There is no absolute certain measurement. Different measurement apparatus

30.56 0.01 kg, 57.8 0.1

A measurement always has

Different people estimate

What are the uncertainties on weights?

What are the considerations for the Lab Glassware?

Graduated 10.00 ml pipitte: