BAHIR DAR UNIVERSITY

BAHIR DAR INSTITUTE OF TECHNOLOGY (BIT)

FACULTY OF CHEMICAL AND FOOD ENGINEERING

PRACTICAL ANALYTICAL CHEMISTRY MANUAL

(CHEM………..)

September, 2017

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Table of Contents
Hazard Symbols...........................................................................................................................................iii
General Laboratory Safety..........................................................................................................................iv
Basic Laboratory Equipments (Glass Wares) ..............................................................................................vi
Laboratory Report Format..........................................................................................................................ix
Solution Chemistry......................................................................................................................................xi
Preparation of Solutions ...........................................................................................................................xiii
Part 1: Titrimetric Methods of Analysis ...................................................................................................... 1
EXPERIMENT 1: standardization of sodium hydroxide solution............................................................. 1
EXPERIMENT 2: Determination of Na2CO3 and NaHCO3 in the Same Solution..................................... 4
EXPERIMENT 3: Determination of Halides Argentometrically................................................................ 6
EXPERIMENT4: Determination of Oxalate Permanganometrically ...................................................... 10
EXPERIMENT 5: Determination of Hardness of Water ......................................................................... 12
Part 2: Selected Gravimetric Method ....................................................................................................... 14
EXPERIMENT 1: Determination of Iron in Ferric Hydroxide ............................................................. 16
EXPERIMENT 2: percentage of water in a hydrate ............................................................................... 18
EXPERIMENT 3: Determination of the Empirical Formula of a Hydrate ............................................... 22
Part 3: Chromatographic Separation ........................................................................................................ 28
EXPERIMENT 1: paper chromatography of dyes .................................................................................. 29
EXPERIMENT 2: thin layer chromatography ......................................................................................... 31
EXPERIMENT 3: Column Chromatography............................................................................................ 34
Part 4: Selected Experiments on Electro Analytical Methods .................................................................. 38
EXPERIMENT 1: Molar Conductivities of Aqueous Electrolytes ........................................................... 38
EXPERIMENT 2: Determining the PH of Various Substances................................................................ 41
Part 5: Experiments on Spectroscopy....................................................................................................... 43
EXPERIMENT 1: UV/VIS Spectroscopy and Spectrophotometry: Spectrophotometric Analysis of
Potassium Permanganate Solutions. .................................................................................................... 44
Further Readings/ References .................................................................................................................... 1

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HAZARD SYMBOLS

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 GENERAL LABORATORY SAFETY
1. At the outset, learn the location of the nearest eye fountain, tire blanket, and fire
extinguisher. Lear the proper use of each, and do not hesitate to use this equipment should the
need arise.

2. Wear eye protection at all times. The potential for serious and perhaps permanent eye injury
makes it mandatory that adequate eye protection be worn at all times by students, instructors
and visitors. Eye protection should be donned before entering the laboratory and should be used
continuously until it time to leave. Serious eye injuries have occurred to people performing such
innocuous tasks as computing or writing in a laboratory notebook; such incidents usually
result from someone else’s loss of control over an experiment. Regular prescription glasses are
not adequate substitutes for eye protection approved by the office of safety and health
administration (OSHA). Contact lenses should never be used in the laboratory because
laboratory fumes may react with them and have a harmful effect on the eyes.

3. Most of the chemicals in a laboratory are toxic; some are very toxic, and some such as
concentrated solutions of acids and base are highly corrosive. Avoid contact between these
liquids and the skin .in the event of such contact. Immediately flood the affected are with
copious quantities of water. If a corrosive solution is spilled on clothing, remove the garment
immediately. Time is of the essence; do not be concerned about modesty.

4. Never perform unauthorized experiment. Unauthorized experiments are grounds for

disqualification at many institutions.

5. Never work alone in the laboratory; be certain that someone is always within earshot.

6. Never bring food or beverages in to the laboratory .do not drink from laboratory glassware.
Do not smoke in the laboratory.

7. Always use a bulb or other device to draw liquids in to a pipette: never use your mouth to
provide suction.

8. Wear adequate foot covering (no sandals).confine long hair with a net. A laboratory coat or
apron will provide some protection and may be required.

9. be extremely tentative in touching objects that have been heated; hot glass looks just like cold
glass.

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10. Always fire-polish the ends of freshly cut glass tubing. Never attempt to force glass tubing
through the hole of stopper .instead, make sure that both tubing and hole are wet with soapy
water. Protect your hands with several layers of towel while inserting glass in to stopper.

11. Use fume hoods whenever toxic or noxious gases are likely to be evolved. Be cautious in
testing for odors: use your hand to waft vapors above containers toward your nose

12. Notify your instructor immediately in the event of an injury.

13. dispose of solutions and chemicals as instructed. It is illegal to flush solutions containing
heavy metal ions or organic liquids down the drain in many localities; alternative arrangements
are required for the disposal of such liquids.

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BASIC LABORATORY EQUIPMENTS
(GLASS WARES)

vi
vii
viii
 LABORATORY REPORT FORMAT
The purpose of a scientific report is to convey information. This is true of all sub-disciplines of
chemistry, and more generally true all scientific disciplines. A laboratory report a summary of:
(a) why undertook this research; (b) what you expected to find ;(c) how you actually did it (d)
what you did find (f)how you interpreted the result and (g) the theoretical and practical
implications of the conclusions you drew from those results.

A scientific report consists of several sections. The recommended order of presentation is

indicated in this section.

Cover page of your report should include

 Name of the university, the collage and the department

 Course title: may include the course code

 Experiment title: may include experiment number

 Instructor’s name:

 Student name: may include partner’s Name and ID number

 Section: may include [Link] or group number

 Date

Content of the report body should include:

 Title: deals the topic of the experiment.

 Objective: carefully study and what you are trying to do. One or two sentences are
usually sufficient to indicate the goals of the experiment.

 Theory: this section should provide a brief explanation about the experiment and the
chemical basis of the experiment with balanced equations and any mathematical
equations needed to do the calculations. All terms used in equations should be defined
(not greater than the page).

 Materials and chemicals: this section may include chemical reagents and material
indicated in the manual or other reagents or materials used in the experiment but not in
the manual.
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 Procedure: do not make the list of steps. In paragraph form, give a brief synopsis of
the process and detail any abbreviation from the lab book [Link] should be in the
past passive tense.

 Observation and data: use tables and [Link] them carefully and clearly, make
sure that the units are included. Write every physical change you observed in the
experimental procedure.

 Calculation: you must take a set of your data and show a sample calculation in detail.
As you do the calculation, be sure to carry all units through to be sure the results have
the correct units. This also allows the instructor to identify any errors you have made.

 Result and Discussion: this section should contain statistical analysis of data in terms
of mean, relative standard deviation etc. and possible source of errors.

 Conclusion: this section summarizes your main results, and includes some comments
on possible source of errors.

 Answer for questions: include answer for questions whenever necessary.

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 SOLUTION CHEMISTRY
Solutions: are homogeneous mixtures that contain components of solutes and solvents.

Solute: .is a substance that can be dissolved by a solvent and exists less amount in a solution.

Solvent: is a substance that dissolves a solute and exists large in amount.

Types of solutions

Based on the amount of solute in a solution:

1. Unsaturated solution: before saturation point is reached it contains less solute than it has the
capacity to dissolve or contains less amount of solute than required for saturation.

2. A saturated solution: is a solution in which the dissolved and undissolved solutes are in
dynamic equilibrium or contains as much solute as that can be dissolved in a given amount of
solvent.

3. Supper saturated solution: contains more solute than is present in a saturated solution or
contains extra amounts of solute than that of required for saturation that is some solutes
remaining as undissolved.

Concentrations of solutions

Concentration is the amount of solute/solvent found in a given solution and that can be
expressed as qualitative and quantitative terms.

Qualitative terms:

Diluted solution: contains less amount of solute than other solution of the same type.
Concentrated solution: contains large amount of solute than other solution on the same type.

Quantitative terms: expressed using different concentration expressions. These are;

1. Percentage concentration: expresses the percentage of solute/solvent/mass/volume in the

solution.

a. %weight/weight= (wt of solute /wt of solution)*100

b. %weight/volume= (wt of solute (g) /volume in (ml) solution)*100

c. %volume/volume= (volume of solute /volume of solution)*100

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2. Molarity (M): the ratio of moles of solute per litre of solution.

M=moles of solute/volume (L) of solution

3. Molality (m): the ratio of moles of solute to mass of solvent in kilogram.

m=moles of solute/mass in Kg of solvent

4. Normality: expressed as number of equivalents of solute per litre solution.

N=no of eqvt of solute/volume in litre solution =mole of solute *z/ volume (L) solution=M*Z

Since, no of eqvt=given mass/eqvt weight

Eqvt weight=molar weight/z , z is positive charge in the formula

Mole=given mass/molar mass

5. Mole fraction(x): the ratio of mole of one component to the solution of all moles of each
component in the solution. E.g. if a solution composition of two compounds A&B,

XA=mole of A/mole of A +mole of B , XB=mole of B/mole of A+ mole of B

Note: the sum of XA+XB+……………+Xn=1

6. Parts per million (ppm) & parts by billion (ppb)

Ppm = (mass of analyte/mass of sample)*106 , ppb=(mass of analyte/mass of sample)*109

[Link] ethanol solution is prepared by dissolving 10ml of

ethanol,C2H5OH(density=0.789g/ml) in a sufficient volume of water to produce 100ml of
solution in a density of 0.982g/ml. what is the concentration of the solution in

a. %wt/wt b. %wt/v c. %v/v d. M e. m f. mole fraction

Question2. Calculate the mole fraction of water and ethanol in a mixture of equal masses of
water and ethanol.

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Preparation of Solutions

Preparation of solutions from solids;

Follow the following procedures

 wash & clean the apparatus carefully

 calculate the mass of solute needed
 add the solute to volumetric flask & add some solvent
 stir& dissolve the solute
 adjust the volume up to the mark by adding solvent

[Link] can you prepare 0.1MNaOH aqueous solution in 100ml?

Preparation of solutions from liquids

Acids, organic solvents,& some bases are found in liquid state in nature.

You have to notice the density /specific gravity, %composition on the container of the stock/
concentrated solution. In this case using dilution law

Dilution law: is preparation of more diluted solution from concentrated solution.

Mathematically, C1V1=C2V2, C1=the con of stock/concentrated solution

V1=the volume needed from stock solution

C2=con of the solution to be prepared

V2=volume of the solution to be prepared

C1 in terms of molarity & normality obtained by:

M=C1= (density/specific gravity*(%composition/100)*1000)/molar weight

N=C1= (density / specific gravity*(%composition/100)*1000)/eqvt weight

[Link] can you prepare 800ml of 0.4M OF NH3 from a concentrated solution that has a
specific gravity of 0.9 & is 27%wt/wt NH3?

[Link] concentration of C29H60 in summer rain water is 34 ppb. What is the molarity of
this alkenes? (Mwt C29H60=408.8g/mol).

xiii
[Link] the molar and molal concentrations of HCl in a reagent labeled 37%wt/wt HCl
and density of 1.188g/ml. the density of the substance is the mass per unit volume.

[Link] does it mean 98%wt/wt, 85wt/v, 75v/v, and how to make solutions from these
concentrations?

Answers: Q1 a.8.3% b.7.89% c.10% d.1.715M e.1.899m f. Xethanol=0.033,

Xwater=0.967)

[Link]=0.28 , Xwater=o.719

Q3. .0.4g.

Interpretation; adding 0.4g of NaOH in to volumetric flask and dissolved by water and then
adding water up to the mark of the volume.

Q4.M=C1=14.29M , V=22.39ml

Interpretation: take 22.39ml of NH3 from the stock solution to volumetric flask and dilute by
water and then adjust the volume up to the mark of 800ml.

Q5. M=8.3*10-8M

Q6.M=12.1M , m=16.1m

Q7.98%wt/wt; 98g of solute in 100g solution

85%w/v: 85g of solute in 100ml solution

75%v/v: 75 ml solute in 100ml solution

Note; adding each mass for solids / volume for liquids to volumetric flask in order to make
appropriate solution.

Errors in analytical results/ in any laboratory work

Error: is the deviation from the true value (U) to measured /mean value(X).

E=|u-x| , %E=(|u-x|/u)*100

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There is no analysis that is free from error completely, but a good analyst knows the error and
try to minimize it. There are three types of errors, these are;
[Link]/ systematic/ regular error

 an error whose source can be predicted from where

 It makes all measured values higher/ lower than the true value
 Possible to minimize & avoid completely
 Affects the accuracy of the measurement
 Recognizable by the lack of agreement b/n measured value & the true value
 Quantifiable by the difference b/n measured value & true value
 Lower or higher values far from the true value equally

2. Indeterminate/ random/ irregular error

 It is impossible to predict the source of this error

 Affects the precision of the measurement
 It oscillates around the true value
 Possible to minimize ,but not avoid completely
 Quantifiable by measuring the spread
 Recognizable by scattering around the mean
 Values are below or above/ oscillates the true value

[Link] error/occasional error

 Causes to be one of the measured values to be an outlier.

 Outlier: the deviation of one quantity from others significantly.

Sources of error

1. methodological errors; slowness of reactions, incompleteness of reactions

 Side reactions; due to presence of interferences, non specificity of reagents.
2. instrumental errors; wrong calibration of instruments
 Voltage interruption
3. personal error; from the fault of analyst
 Carelessness of the analyst

xv
 PART 1: TITRIMETRIC METHODS OF
ANALYSIS
The term titrimetric analysis refers to quantitative chemical analysis carried out by determining
the volume of the solution of accurately known concentration which is required to react
quantitatively with a measured volume of a solution of a substance or an analyte to be
determined. The term volumetric analysis was used for this form of quantitative determination
but it has now been replaced by titrimetric analysis. The solution of accurately known
concentration is called standard solution. In Titrimetric analysis, the reagents of known
concentration is called titrant and the substance being titrated is termed the titrand the process
of adding a measured volume of standard solution to a solution containing a substance to be
determined is called titration.

The standard solution (titrant) is gradually added until chemically equivalent amount is reacted
with the analyte solution (titrand). The point at which the amount of standard solution is
chemically equivalent to that of the analyte is called equivalence point. Therefore, at this point,
number of equivalent of standard solution is exactly equal to number of equivalent of the
analyte solution.

There are several kinds of chemical reactions that show changes which can be detected and thus
utilized for analysis by titration. For the sake of convenience, titrimetric analysis may be
divided in to four classes. These are; acid-base (neutralization), precipitation, complex
formation and oxidation-reduction titrations.

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EXPERIMENT 1: STANDARDIZATION OF SODIUM HYDROXIDE SOLUTION

Objective: to standardize sodium hydroxide using pre-standardized secondary standard

hydrochloric acid

Theory:

Standard solution is a reagent of exactly known concentration that is used in titrimetric

analysis .standard solutions play a vital role in all titrimetric method of analysis. The
concentration is usually expressed in normality and morality. The process by which the
concentration of a solution is determined is called standardization.

The desirable properties of an ideal standard solution for titrimetric method are:

 be sufficiently stable so that it is only necessary to determine the concentration

once;

 reacts rapidly with the analyte so that the time required between additions of
reagent is minimized ;

 reacts more or less completely with the analyte so that satisfactory end points are
realized;

 undergoes a selective reaction with the analyte that can be described by simple
balanced equation;

There are two types of standard substances; primary and secondary standards

Primary standard: is analytically pure substance and prepared by dissolving a known amount of
a primary standard in a suitable medium and diluting to a definite volume, a solution of known
concentration is readily prepared. Compound such as borax, Na2B4O7.10H2O (equiv. wt
190.72),sulphamiacid,NH2SO3([Link].97.098), sodium carbonate,Na2CO3([Link].53.00),
potassium hydrogen iodate, KH(IO3)2 ([Link].389.928),potassium hydrogen phthalate
(KHC8H4O4) ([Link].204.22),oxalic acid,H2C2O4.2H2O([Link].63.05)are common
examples of primary standards

To be a primary standard; a substance should meet the following requirements;

 It should be easily obtained in an analytically pure state (>99.99%);

1
  Stability towards air;

 It is equivalent weight ought to be high in order to reduce the effect of small

weighing error;

 It should be reasonable soluble under the given condition of analysis;

 Ready available at modest cost;

 On titration, no interfering products should be present;

 The primary standard should be colorless, before after titration.

 The prepared standard solution should be one that does not change concentration
with time.

Secondary standard: is a less pure compound that sometimes used in place of primary standard
and whose purity has been established by chemical analysis and serves as the reference material
for titrimetric method of analysis. Compound such as sodium hydroxide (NaOH) or
hydrochloric acid (HCl) cannot be considered as primary standard since their purity is changed
through time.

Chemical and Apparatus

HCl, Na2B4O7.10H2O, NaOH and methyl red, volumetric flask, iron stand, clamp, conical
flask, beaker, pipette, balance and burette.

Procedures

1. prepare an approximately 0.2N solution of hydrochloric acid by diluting 1ml of

concentrated hydrochloric acid (usually between 10N and 11N) to 100ml volumetric flask.

Caution: if any acid is split on your skin or eyes, thoroughly rinse the affected area with
water, or dilute solution of sodium hydrogen carbonate .for sever case consult a medical
doctor.

2. Prepare 0.1N of borax solution. Weight accurately 1.9072g of borax

(Na2B4O7.10H2o).transfer to a 100ml volumetric flask, add about 30ml of distilled water
and shake until dissolved. Adjust the volume to the mark and mix well.

2
3. Pipette exactly 25 ml of the borax solution into 250 ml conical flask, and 2-3 drops of
methyl red indicator and titrate it with the hydrochloric acid until the first red tint color
appears indicating the end point of titration .repeat the titration three times.

4. Calculate the concentration of hydrochloric acid using the following expression

2(NHClXVHCl)=NboraxXVborax=N1V1=N2V2, where N1 and N2 normality’s of HCl and
borax respectively and V1 and V2are volumes of HCl and borax respectively needed to
reach the end point. N.B.N1V1=N2V2

5. Using the standardized secondary standard hydrochloric acid, now standardize the sodium
hydroxide as follows.

6. Dissolve 4.5g of sodium hydroxide in previously boiled and cooled distilled water

7. Transfer it to 100 ml volumetric flask and dilute it to the mark.

8. Pipette out 25 ml of NaOH solution in to 250 ml volumetric flask.

9. Add 2-4 drops of methyl red indicator.

10. Titrate it with the standardized HCl solution to the first appearance of persistent red tint
color. Calculate the normality of NaOH solution using the above expression.

Post Lab Questions

1. How would it affect your results if you used a beaker with residual water in it to
measure out your standardized sodium hydroxide solution?
2. How would it affect your results if you used a wet Erlenmeyer flask instead of a dry one
when transferring your acid solution from the volumetric pipette?
3. How do you tell if you have exceeded the equivalence point in your titration?
4. Vinegar is a solution of acetic acid (CH3COOH) in water. For quality control
purposes, it can be titrated using sodium hydroxide to assure a specific %
composition. If 25.00 mL of acetic acid is titrated with 9.08 mL of a standardized 2.293
M sodium hydroxide solution, what is the molarity of the vinegar? What is the %
composition (wt/wt %)?
5. How will you know when your titration is finished?

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EXPERIMENT 2: DETERMINATION OF NA2CO3 AND NAHCO3 IN THE SAME
SOLUTION

Objective: To determine the amount of Na2CO3 and NaHCO3 in the same solution by the
existence of two equivalent points on titration curve.

Theory

If the mixture of Na2CO3 and NaHCO3 is titrated with standard HCl solution in the presence of
phenolphthalein indicator, the light pink color disappears when Na2CO3 is changed in to
NaHCO3 .the volume of HCl solution (say Vm ml) used up will account for half neutralization of
Na2CO3.

Na2CO3 + HCl → NaCl + NaHCO3

To the same titration mixture, if methyl orange indicator is added and the titration is continued,
the color is changed from strew yellow to pale red indicates total neutralization.

NaHCO3 + HCl → NaCl + CO2 + H2O

The volume of hydrochloric acid required for this reaction (say Vn ml) will account for the
second half neutralization of Na2CO3 and complete neutralization of NaHCO3 originally
present in the given analyte solution. From the volumes of hydrochloric acid solution (V m and
Vn ml) and reaction (1) and (2), it is possible to calculate the amount of of Na2CO3 and
NaHCO3 in the given solution.

Chemical and apparatus: borax solution, hydrochloric acid, methyl red, phenolphthalein and
methyl orange, analyte solution containing Na2CO3 and NaHCO3, iron stand, clamp, volumetric
flask, conical flask, pipette, balance and burette.

4
Procedures

i) Preparation of 0.1N borax, the same as the procedure given in experiment 1

ii) Standardization of HCl solution, the same as the procedure given in the experiment

iii) Estimation of Na2CO3 and NaHCO3

1. Pipette out 25 ml of the analyte solution containing Na2CO3 and NaHCO3 in to a clean
conical flask. Add 2-3 drops of 0.5%phenolphthalein indicator solution and titrate it the
previously standardized HCl solution taken in to burette until the pink color just
disappeared. Note down the volume of HCl required for this end point (say Vm ml).

2. Add 3 drops of 0.05% solution of methyl orange indicator to the same titration mixture
and resume the titration. The end point is indicated by the color change from strew
yellow to pale red .note down the volume of HCl required for this end point ( say V n
ml).

3. Repeat the titration three times. Calculate the normality’s of Na2CO3 and NaHCO3 and
the amount in gram per liter.

Post Lab Questions

1. Explain the differences between equivalent point and end point briefly.
2. Why does phenolphthalein change color around pH 9.0 and methyl red around pH 5.0?
3. Experimental yield is calculated by taking the value you determine in your experiment,
divide by the value you expected, and multiply by 100%. How much NaCl did you
obtain in your experiment? How much NaCl should you have gotten (based on your
mass of NaHCO3 or Na2CO3 you used)? What is your yield?
4. What are some sources of error that would cause your experimental yield to be less than
100%?

5
EXPERIMENT 3: DETERMINATION OF HALIDES ARGENTOMETRICALLY

Objective: estimation of the amount of chloride in a given solution.

Theory

Titrimetric methods in which AgNO3 used as titrant are known as argentometric methods. They
can be direct method, in which the titrant is directly added to rest a solution until the end point
is reached or indirect method, in which excess of the titrant (AgNO3) solution is added to halide
solution and the excess AgNO3 solution is back titrated with a different standard solution like
NH4SCN or KSCN

Three methods have been use to determine halides Argentometrically.

1. Mohr’s method:

This method is direct method using K2CrO4 as indicator. The method is based on the fact that
the solubility of AgCl is less than that of Ag2CrO4. As such, the brick red colored precipitate of
Ag2CrO4 is formed only after the chloride in analyte solution is completely precipitated, thus
indicating the end point. However the amount of chromate ion in the solution has significant
effect on the accuracy of the end point. Usually in this method, indicator blank titration is
performed to obtain better result. The reaction is:

Cl- + Ag+ → AgCl(s) (white)

2 Ag+ + CrO4-2 → Ag2CrO4 (brick red)

2. Fajan’s method:

This is also a direct method, using an organic adsorbing agent as indicator .in this method, an
adsorbing indicator like dichloroflouroscein gets adsorbed with excess Ag+ ions, just after
equivalent point and shows distinct color change.

3. Volhard’s method: this is indirect method (back titration method) using excess
AgNO3, which is back titrated by thiocyanate using ferric alum (Fe3+) as indicator. The
chloride in analyte solution is precipitated as AgCl and excess AgNO3 on titration with
SCN- forms AgSCN. As the solubility of AgCl is more than that of AgSCN, the AgCl
precipitate is filtered of, or is coated with an organic reagent like nitro benzene, before
titrating with thiocyanate solution .just after the end point, Fe3+ forms a blood-red
colored complex with SCN. The reaction is:
6
 Ag+ + SCN- →AgSCN (white

Fe3+ + 6SCN- → [Fe (SCN) 6]-3 (blood –red)

Chemical and apparatus

NaCl,AgNO3,K2CrO4, dichloroflouroscein ,dextrin and unknown chloride solution ,burette,

conical flask, pipette, iron, stand, clamp, volumetric flask and measuring cylinder.

Procedures

This experiment is performed in the following steps:

1. Preparation of standard 0.1N NaCl solution. Weight accurately 0.585 g pure ([Link])
dry NaCl. Transfer to a 100mL volumetric flask, add about 30 ml of distilled water and
shake until it dissolved. Adjust the volume to the mark and mix well.

2. Standardization of AgNO3 solution (0.1N).

i. Pipette out 10mL of standard 0.1N NaCl solution in to a clean conical flask
and add1.0 ml (0.01) K2CrO4 as indicator.

ii. Titrate slowly with AgNO3 solution taken in burette until brick-red color of
precipitate is formed which indicates the end point .note the volume of AgNO3
(say v1mL) required, and repeat the titration three times.

iii. Determine black indicator with 1mL of K2CrO4 solution and 50 ml of distilled
water mixture, which can be titrated with AgNO3 solution until color
matching with that observed in procedure (ii).the volume of AgNO3 is (say V2
mL) is to be subtracted from the earlier titer valve (V1), to determine the
volume consumed by the analyte only. Calculate the normality of AgNO3?

3. Standardization of KSCN Solution.

Pipette out 10mL AgNO3 solution into a clean conical flask and 1-2 drops of ferric alum
solution (0.005M) as indicator. Titrate against KSCN solution taken in burette, until blood-red
color appears which indicate the end point. Repeat the titration to obtain equal titer values and
calculate the normality of KSCN.

7
 4. Estimation of chloride in the given solution as unknown.

A. Mohr’s method
i. Pipette out 10 ml of unknown NaCl solution in to clean conical flask and add 1.0
ml K2CrO4 (0.01) as indicator.
ii. Titrate slowly with standardized AgNO3 solution taken in burette until brick- red
color of precipitate is formed which indicates the end point. Note the volume of
AgNO3 (say V1 ml) required, and repeat the titration three times.
iii. Determine indicator blank with 1 ml of K2CrO4 solution and 50 ml of distilled
water mixture, which can be titrated with AgNO3 solution until color matching
with that observed in procedure ii is observed .the volume of AgNO3 is (say V2
ml) is to be subtracted from the earlier titer value (V1), to determine the volume
consumed by the analyte only.
iv. Calculate the normality of NaCl and amount of chloride in g/L?

Where: VAgNO3= (V1-V2) ml, NAgNO3=known from (ii)

B. Volhard’s method

i. Pipette 10 ml of analyte chloride solution into a 400 ml beaker .add 3 ml of concentrated

HNO3 and standard AgNO3 solution until all the chloride is precipitated and 5 ml is in excess
(say 15 ml). Note down the exact volume of AgNO3 added. Stir well, filter through a whattman
filter paper grade 1, and wash the precipitate with (1:100) HNO3 solution.

ii. Collect the filtrate and washing into clean conical flask, and add 1-2 drops of ferric alum
solution (0.005M) as indicator and titrate against KSCN solution taken in burette until blood –
red color indicates end point.

C. indicator method (Fajan’s method)

i. Pipette out 15 ml of analyte solution in to clean conical flask .add 3-4 drops of
dichloroflouroscein indicator and 0.1 g dextrin (stabilizer for colloidal AgCl
precipitate). Titrate rapidly with AgNO3 solution from burette until permanent pink
color indicates the end point. Repeat the titration for constant titer valves and
calculate the amount of chloride solution in the given solution and compare the three
methods.

8
 Post lab questions

1. 25 ml of 0.05 N NaCl required 20 ml of AgNO3 solution for the end point in the presence of
K2CrO4 indicator. The same AgNO3 solution was used to estimate chloride in KCl solution,
following Mohr’s method, in which 25 ml of the KCl consumed 22.5 ml of AgNO3 solution.
Calculate the normality of the link AgNO3 solution and the concentration of KCl solution.

2. 25 ml of 0.08N AgNO3 required 24 m of KSCN solution to attain end point in the presence
for ferric alum indicator. In a separate experiment 20 ml NaCl solution of unknown
concentration was treated with 25 ml of the 0.08 N AgNO3 solution and the resultant
mixture was titrated with the same KSCN solution used in the previous experiment
following Volhard’s method. 10m L of KSCN solution was required for the titration.
Calculate the concentration of KSCN solution and estimate the amount of chloride in the
given solution.

9
EXPERIMENT4: DETERMINATION OF OXALATE PERMANGANOMETRICALLY

Objective: estimation of the amount of oxalate in a given solution using KMnO4

Theory:

Titration involving KMnO4 as oxidizing agent are called permanganometric titration.

Permanganate ion is one of the most powerful and widely used oxidizing agents. depending on
the pH of the medium MnO4- can undergo 5e.4e,3e and 1e reaction and can be reduced to
+2,+3,+4 and+6 oxidation states respectively. In solution that are 0.1N or greater in mineral
acids MnO4- is reduced to Mn (||).

MnO4- + 8H+ + 5e- → Mn2+ + 4H2O

Under these conditions it can oxidize oxalic acid or sodium oxalate. H2SO4 is most suitable to
provide the required H+

2MnO4- + 5H2C2O4 + 6H+ → 2Mn2++ 10CO2 + 8H2O

2MnO4- + 5C2O42- +16H+ → 2Mn2+ + 10CO2 +8H2O

HCl is not advisable as the following reaction is expected to occur.

2MnO4- + 10Cl- + 16H+ → 2Mn2+ + 5Cl2 + 8H2O

The reaction proceeds slowly at room temperature and hence the titration is initiated around
600C. Once reasonable concentration of Mn2+ is built up, the reaction becomes rapid due to
Mn2+ catalysis. As KMnO4 is purple in color, the titration do not need an external indicator for
detecting the end point .however , when dilute KMnO4 is used (0.1 *10-6N), oxidation –
reduction indicators such as sodium diphenylamine sulphate, N-phenylanthranilic acid and
ferroin may be used to enhance the intensity of color change at the end point.

Chemical and apparatus

H2C2O4.2H2O, Na2C2O4, KMnO4, H2SO4, volumetric flask, conical flask, measuring cylinder,
beaker, iron stand, clamp, burette and pipette.

10
Procedures

1. Preparation of standard oxalic acid solution

0.63g oxalic acid dehydrated (H2C2O4.2H2O) dissolve in distilled water to make 100 ml
solution in volumetric flask that will be 0.1 N solutions
2. Standardization of KMnO4 solution

Pipette out 10mL standard oxalic acid solution in to a clean conical flask and add 10 ml of 4N
[Link] the contents to 600C (appearance of first bubble) and titrate against KMnO4 solution
taken in the burette until appearance of faint pink color indicates the end point.

Perform the titration slowly with constant shaking of the titration mixture. Note the volume of
KMnO4 and repeat the titration for equal titer values. Calculate the normality of KMnO4
solution.

3. Estimation of Na2C2O4 in the given solution

Pipette out 10mL Na2C2O4 solution into a clean conical flask and add 10mL of 4N H2SO4.

Heat the contents to 60oc (appearance of first bubble) and titrate it against KMnO4 solution
taken in the burette until appearance of faint pink color indicates the end point. Perform the
titration slowly with constant shaking of the titration mixture. Note the volume of KMnO 4 and
repeat titration for equal titer values. Calculate the normality of KMnO4 solution .calculate the
normality and molarity of Na2C2O4solution.

Post lab. Questions

1. If 6 ml of 3M H2SO4 solution were diluted to 250 ml, what would be the

concentration of H2SO4 in the final solution?
2. Suppose 0.125 grams of Na2C2O4 are dissolved to make 250.0 ml of solution. How
many moles of Na2C2O4 are present in each 1.00 ml of this solution?
3. Suppose 1.00 ml of this solution is titrated with a KMnO4 solution in order to
determine the concentration of the KMnO4. The end point occurs after the addition of
145 ul. What is the molar concentration of the KMnO4 solution?
4. Suppose 0.125 grams of an unknown mixture of Na2C2O4 and KCl (an inert
additive) are dissolved in 250 ml of Solution. If a 1.00 ml sample of this solution
requires 93.0 ul of the same KMnO4 solution, what is the % sodium oxalate in the
unknown?

11
EXPERIMENT 5: DETERMINATION OF HARDNESS OF WATER

Objective: to determine the hardness of water complexometrically using EDTA

Theory

The presence of metal ions like Mg2+and Ca2+ interfere with the ability of water and cause
hardness. The process adapted to remove hardness is called softening of water .EDTA (ethylene
diaminetetraacetic acid) is versatile reagent, widely used to determine the extent of hardness in
water. This is due to the ability of EDTA to form stable 1:1 complexes with various metal ions
under suitable pH. There are six potential metal binding centers in EDTA which includes two
nitrogen and four acetate oxygen centers caused by the deprotaonation of four carboxylic acid
functions.

In view of low solubility of EDTA, it is generally used as disodium salt

Na2H2C10H12O8N2.2H2O (FW=372.24g/mol) in complexometric titrations. Metal ion indicators
like eriochrom black T, muroxide, xylonol orange, fastsulphon black,etc are used in these
titrations. The indicators also form complexes with metal ion which get displaced by ethylene
diamine tetra acetate (Y2-) during titration under suitable conditions. This is due to the
difference in the stabilities of metal-EDTA and metal indicator complexes.

The bound and unbound indicator species impart different colors to titration media, these
resulting in distinct color changes at the end point of titration.

H2Y2- + Mg-In- → Mg Y2-+ H In2-+ H+

Wine red blue

In3- and HIn2- are the forms of EBT in metal bound and metal unbound forms with wine red and
sky blue colors, respectively.

Chemical and apparatus:

MgSO4.7H2O,disodium salt of EDTA, aqueous ammonia, NH4Cl,EBT,volumetric flask,

conical flask, measuring cylinder, iron stand, clamp, beaker, burette and pipette.

12
Procedures

This experiment is conducted in three parts.

i. Preparation of standard (0.005M)Mg2+ solution

MgSO4.7H2O (FW=246.47) can be used as primary standard. 1.232 g of this salt
when dissolved in water to make 100mL solution will give 0.05M solution.

ii. Standardization of EDTA ( disodium salt dehydrate FW=372.24)

Pipette out 10mL of standard Mg2+ ion solutions into a clean conical flask ,add
3mL ammonia-ammonium chloride buffer(pH=10),five drops of EBT indicator
solution and heat the contents on water bath up to 40 oC . Titrate with EDTA
solution until the color changes from wine red to sky blue indicating the end point.
iii. Determination of total hardness of water
Pipette out 10mL of hard water into a clean conical flask and titrate against standard
EDTA solution, following the same procedure as given in (ii). Repeat the titration
for equal titer value and calculate total hardness of water in moles/litre.

Post Lab Questions

1. 10mL of Mg+2 ion solution was mixed with 6mL of 0.05M Ca+2 ion solution and the
mixture was diluted to 25 ml. This 4 ml of (NH3 + NH4Cl) buffer (pH=10) was
added and titrated with EDTA solution in the presence of EBT indicator. The
volume of EDTA solution required for titration was 20 ml. What is the
concentration of EDTA?
2. 10 ml of a solution containing Ca+2 was mixed with 2 ml of0.1N Mg+2 solution and
titrated with the same EDTA solution as in (1) under similar conditions .if it takes
14.8 ml of EDTA to reach end point, what is the concentration of Ca+2 solution?

13
 PART 2: SELECTED GRAVIMETRIC
METHOD
Gravimetric analysis is a class of analytical techniques used to determine the mass or
concentration of a substance by measuring a change in mass. The chemical we are trying to
quantify is sometimes called the analyte. Gravimetric analysis describes a set of methods
in analytical chemistry for the quantitative determination of an analyte based on the mass of a
solid.

In most cases, the analyte in solution is first converted to a solid by precipitation with an
appropriate reagent. The precipitate can then be collected by filtration, washed to remove
impurities, dried to remove traces of moisture from the solution, and weighed. The amount of
analyte in the original sample can then be calculated from the mass of the precipitate and its
chemical composition. Gravimetric analysis may be classified as: precipitation gravimetry,
volatilization gravimetry, and electro gravimetry. Note: due to the take of materials, we will do
only precipitation and volatilization gravimetry.

Volatilization gravimetry involves separating components of our mixture by heating or

chemically decomposing the sample. The heating or chemical decomposition separates out any
volatile compounds, which results in a change in mass that we can measure. Precipitation
gravimetry uses a precipitation reaction to separate one or more parts of a solution by
incorporating it into a solid. The phase change occurs since the analyte starts in the solution
phase and then reacts to form a solid precipitate. The solid can be separated from the liquid
components by filtration. The mass of the solid can be used to calculate the amount or
concentration of the ionic compounds in solution. The steps that are followed during
gravimetric analysis are: preparation of the solution, precipitation, digestion, filtration, washing,
drying, igniting and calculation.

A few additional rules of thumb for a successful quantitative precipitation are:

 The precipitate must be large enough to settle out of solution

 The precipitate must be large enough to be filtered
 The precipitation process must be complete
 The precipitate must be pure
 The precipitate must be stable when dry, or produce a stable product upon ignition

14
The results of a gravimetric analysis are generally computed from two experimental
measurements: the mass of sample and the mass of a product of known composition that is
chemically related to the sample. Relationship between mass of the precipitate and the mass of
analyte is established through gravimetric factor. Gravimetric factor is a numerical factor that
is obtained from stoichiometric relationship between the precipitate and the analyte and it used
to convert the weight of the precipitate to the corresponding weight of the analyte.

a (analyte) b(precipitate)

Where a is stoichiometric coefficient of the analyte and b is stoichiometric coefficient of a

precipitate. The gravimetric factor (GF) is given by:

GF= a x Fwt of the analyte/ b x Fwt of the precipitate

Finally, mass of analyte = mass of precipitate x GF

Example of a Gravimetric Analysis

A 0.5962 g sample of iron ore is dissolved in perchloric acid (HClO4). All iron present is
oxidized to Fe3+. The solution is filtered to remove solid matrix materials and made basic with
addition of ammonium hydroxide. The iron precipitates as the Fe(OH)3 .xH2O gel. The
precipitate is collected in a cistern crucible and ignited to produce Fe2O3. What is the wt. % of
iron in the sample if the analysis produced 0.3210 g Fe2O3?

The overall reaction is

From this we derive the gravimetric

factor relating weight of final material to the weight of iron analyte

The weight % of iron in the ore is obtained from the mass of iron and the sample weight,

15
EXPERIMENT 1: Determination of Iron in Ferric Hydroxide

Objective: to determine the mass of iron in ferric hydroxide

Theory

Iron in ferric salts is determined by precipitating it as Fe(OH)3 using NH4OH as precipitant.

Fe3+ + 3NH4OH Fe (OH) 3 + 3NH4+

The Fe (OH) 3 is ignited and finally weighed as Fe2O3

Fe (OH) 3 Fe2O3 + 3H2O

The analyte solution of ferric salt is prone to hydrolysis when heated, even in the absence of the
precipitant.

Fe3++ 6H2O Fe (OH) 3 + 3H2O

The slimy gelatinous precipitate of ferric hydroxide forms at the bottom and on the sides of the
beaker on heating the solution, making it difficult to handle during the analysis. As hydrogen or
hydronium ions are produced as a result of hydrolysis, acidification of the analyte solution can
express hydrolysis and prevent premature formation of the precipitate. During the process of
precipitation ammonium salt is formed that serves as a coagulant for the amorphous ferric
hydroxide precipitate in the colloidal solution. The most common contaminants that go with this
gelatinous precipitate are adsorption co-precipitants.

The precipitate should be washed free from chloride, because volatile ferric chloride is likely to
be formed during ignition resulting in a negative error of the estimation. During ignition, if the
precipitate is excessively heated, the Fe2O3 could be partially reduced, making the stoichiomery
of the final product unpredicted.

Chemicals and apparatuses

Analyte solution, ferric salt, HNO3, NH4OH, AgNO3, NH4NO3, measuring cylinder, beaker,
filter paper, funnel, hotplate, stirrer, crucible, desiccators, and furnance.

16
Procedures

 Pipette out 10ml of the given ferric ion solution into a beaker. Acidify with 2ml of conc.
HNO3.

 Heat the solution over a small burner flame. Don’t boil it.

 Add 10% ammonia solution to the hot solution while stirring until it smalls faintly but
distinctly ammonical. Stir the contents of the beaker vigorously with the glass rod, and
add about 50ml of hot distilled water to diminish adsorption. Stir the liquid well once
again and allow the precipitate to settle for 5 minutes.

 Then check for complete by careful addition of 1-2 drops of NH4OH and filter at once to
avoid extraction of silica from the beaker by the alkaline mother liquor.

 Decant the clear solution in to filter paper. Wash the precipitate several times by
decantation with hot 2% NH4NO3 solution. Then transfer the precipitate quantitatively
to the filter paper and remove particles adhering to the beaker and rod with pieces of
ashless paper, which are transformed on to the filter paper along with the precipitate
partially.

 Continue to wash the precipitate on the filter paper until the chloride ions have been
completely removed i.e. until a portion of the filtrate, acidified with nitric acid, no
longer give turbidity with silver nitrate. The precipitate, filtration, and washing must all
be completed during the same session.

 The filter paper containing the precipitate is carefully transferred to a dry weighed
crucible. It is carefully dried and charred at a low flame. Then the crucible and
precipitate are ignited and heated on a blue flame to a constant weight. Calculate the
amount of iron in the Fe2O3.

Post lab. Questions

1. Why must be a washing liquid generally contain an electrolyte?

2. Why is it not desirable to apply suction filtration on the ferric hydroxide precipitate?

3. In a gravimetric experiment, 0.484g sample of iron ore was decomposed in to a

solution was determined gravimetrically by precipitation it as Fe2O3. The weight on
the precipitate was found to be 0.242g. Calculate %w/w of iron in the sample.

17
EXPERIMENT 2: PERCENTAGE OF WATER IN A HYDRATE

Objectives: to determine the percentage of water in barium chloride dehydrate, to determine

the percentage of water in an unknown hydrate salt, to calculate the water of crystallization for
the unknown hydrate salt.

Theory
A hydrate salt is composed of anions (negative ions) and cations (positive ions) which are
surrounded by and weakly bonded water molecules. Each hydrate salt has a fixed number of
water molecules associated with it, called waters of hydration or water of crystallization. When
a salt holds waters of hydration, we call it a hydrated salt or a hydrate (hydrate from hydor, the
Greek word for water).

Barium chloride dihydrate, BaCl22H2O, has two waters of crystallization, or two waters of
hydration. Other hydrates have waters of hydration ranging from one to twelve. Upon heating,
a hydrate decomposes and produces an anhydrous salt and water (in the form of steam).


BaCl22H2O (s ) BaCl2(s) + 2 H2O (g)

Examples and Problems

Problem 1-Theoretical Percentage

Calculate the theoretical percentage of water in barium chloride dihydrate.

Solution: The formula mass of BaCl22H2O is

Ba 1 x 137.3 = 137.3 amu

Cl 2 x 35.5 = 71.0 amu

H2O 2 x 18.0 = 36.0 amu

244.3 amu

The theoretical percentage of water is found by dividing the water of crystallization mass, 36.0
amu, by the hydrate mass, 244.3 amu.

36.0 amu x 100 = 14.7 %

244.3 amu

Problem 2 – Experimental percentage

18
The experimental percentage of water in a hydrate is found by comparing the mass of water
driven off to the total mass of the compound, expressed as a percentage. A 1.250 g sample of
barium chloride dihydrate has a mass of 1.060 g after heating. Calculate the experimental
percentage of water.

Solution: The mass of water lost is found by difference:

1.250 g – 1.060 g = 0.190 g hydrate anhydrous water salt

The experimental percentage of water is

Mass of water x 100 = % of water

Mass of hydrate

0.190 g x 100 = 15.2% water

1.250 g

Problem 3 – Water of crystallization: Calculate the water of crystallization for an unknown

hydrate that is found to contain 30.6 % water. The formula mass of the anhydrous salt (AS) is
245 amu.

Solution: The unknown hydrate is 30.6 % water. Subtracting from 100 %, the hydrate must be
69.4 % anhydrous salt. If we make the sample 100 g, the mass of water is 30.6 g and the
anhydrous salt 69.4 g. To calculate the moles of water and the anhydrous salt (AS)

30.6 g H2O x 1 mole H2O = 1.70 moles H2O

18.0 g H2O

69.4 g AS x 1 mole AS = 0.283 mole AS

245 g AS

To find the water of crystallization, simply divide the mole ratio of water to anhydrous salt.

1.70 moles H2O = 6.01  6

0.283mol AS

The water of crystallization is always a whole number; therefore the formula for the unknown
hydrate is AS6H2O.

19
Chemicals and apparatuses

BaCl22H2O, crucible, tongs, balance, measuring cylinder, distilled water.

Procedures

In this lab, you will find the experimental percentage of water in BaCl 22H2O, you will also find
the percentage of water in an unknown hydrate and then calculate the water of crystallization
of the unknown (the formula mass will be provided)

A. Percentage of water in Barium Chloride dihydrate

1. Obtain a porcelain crucible from the stockroom, rinse with water, and heat on a clay
triangle for five minutes over a direct flame to remove any surface moisture. Use crucible
tongs when handling hot crucibles.
2. Weigh the cool, dry crucible and lid. Record the weight on the data sheet.
3. Place between 1.0 and 1.5 g of BaCl22H2O in the crucible and reweigh the crucible, lid
and contents. Record the mass on the data sheet. Calculate the weight of the hydrate.
Record the mass of the hydrate on the data sheet.
4. Place the crucible back on the clay triangle with the lid almost covering the crucible. Heat
gently at first, then with a hot flame for 10 minutes. Allow the crucible to cool until it is
cool to the touch. Weigh the crucible and lid and record the mass on the data sheet.
5. Complete the calculations to determine the percent water in the compound. Show the
calculations on the data sheet.
6. Dispose of the BaCl22H2O. Repeat the procedure as Trial 2 on the data sheet.

A. Percentage of water in an Unknown Hydrate.

1. Obtain an unknown hydrate from the stockroom and record the unknown number on the
data sheet.
2. Repeat the above procedure (Part A) and report the average percentage of water in the
unknown hydrate.
B. Water of Crystallization in an Unknown Hydrate.
1. Get the formula mass of the unknown hydrate from the stockroom, and calculate the
water of crystallization. Show calculations on the data sheet.

20
DATA SHEET

A. Percentage of Water in Barium Chloride Dihydrate

Trial 1 Trial 2

(1) Mass of crucible and lid _____________g ______________g

(2) Mass of crucible, lid and hydrate _____________g ______________g
(3) Mass of hydrate (2) – (1) _____________g _______________g
(4) Mass of crucible, lid and ____________ g _______________g
Anhydrous salt (after heating)
(5) Mass of water ____________ g _______________g
Show calculations for the experimental percentage of water for Trial 1
Percentage of water in BaCl2H2O _________________% __________________%
Average ________________%
B. Percentage of Water in an Unknown Hydrate

Unknown number _________

Trial 1 Trial
(1) Mass of crucible and lid _____________g ______________g
(2) Mass of crucible, lid and hydrate _____________g ______________g
(3) Mass of hydrate (2) – (1) _____________g _______________g
(4) Mass of crucible, lid and ____________ g _______________g

Anhydrous salt (after heating)

(5) Mass of water ____________ g _______________g

Show Calculations for the experimental percentage of water for Trial 1

Percentage of water in unknown _________________% __________________%

Average ________________%

A. Water of Crystallization in an Unknown Hydrate

B. Formula mass of anhydrous salt (from stockroom) ______________ amu
C. Percentage of water ______________ %
D. Percentage of anhydrous (AS) ______________%
Calculations for the water of crystallization

21
E. Water of crystallization __________________
Formula of unknown hydrate AS H2O

Post lab. Questions

1. When a 1.000 g sample of CuSO4 ·5 H2O(s) was heated so that the waters of hydration
were driven off, the mass of the anhydrous salt remaining was found to be 0.6390 g.
What is the experimental value of the percent water of hydration?
2. A hydrate has the formula of MgSO4 .7H20. What is the percent water in this hydrate?

3. Calculate the percent hydration for barium chloride dihydrate, BaCl2 ·2 H2O(s).
(Atomic masses: H = 1.01; O = 16.00; Cl = 35.45; Ba = 137.33)

EXPERIMENT 3: DETERMINATION OF THE EMPIRICAL FORMULA OF A

HYDRATE

Objective: to determine the empirical formula a hydrate

Theory
When aqueous solutions of inorganic salts are prepared, the ions are stabilized by the process of
hydration. Each negative ion (anion) attracts the positive end of water molecules while each
positive ion (cation) attracts the negative ends. The positive ions, which are usually smaller
than the negative ions show this effect to a greater extent. This results in hydrated cations which
may have considerable stability, especially for the di- and tri-valent transition metal ions. When
aqueous solutions of various salts are concentrated, the forces between water molecules and the
hydrated cations are of sufficient strength that the water molecules are retained in the crystal as
the solid salt separates from solution. These hydrates can be isolated, and have definite
compositions and structures. An example of a hydrate salt is CoCl 26H2O (cobalt (II) chloride
hexahydrate) the cation of which is shown below.

In this salt, there is one [Co{H2O}6]2+ ion and two Cl—ions present. The
cation is bonded to six water molecules in an octahedral array. It should be emphasized that the
lines in the figure therefore do not necessarily represent bonds, they merely indicate the shape
of the ion. Compounds such as this in which a salt and water are combined in definite

22
proportions are known as hydrates, the water associated with the salt is called water of
hydration.

The water bound in these hydrated salts can often be removed at low temperatures
(about 120C) in most cases, to produce water and the anhydrous salt.

CoCl26H2O  CoCl2 + 6 H2O

The percentage of water in a hydrate can usually be determined by heating a known amount of
hydrate until dehydration occurs. The mass of the hydrate equals the mass of the anhydrous salt
plus the mass of the water driven off. The percentage of water in the hydrate can therefore be
calculated:

% = 100
ℎ

Some hydrates lose their waters of hydration readily, simply by exposing them to the
atmosphere. This process is known as efflorescence. For example, when a sample of
MgSO47H2O (known as Epson salt) is placed on a watch glass on the laboratory bench, the
following process occurs:

MgSO47H2O (s)  MgSO4 (s) + 7H2O (g)

This change produces seven molecules of water vapor per molecule of the hydrate. If the
process is carried out in a closed container, the pressure of the water vapor (vapor pressure) can
be measured. For this salt, the vapor pressure equals 11.5 torr at 25C. Some anhydrous
inorganic salts, such as CaCl2 and MgSO4 are hygroscopic that is when exposed to the
atmosphere, they absorb water to form hydrates. Such salts are extensively used as drying
agents or desiccants. Some compounds of this type actually absorb enough water to dissolve
forming a solution. This process is termed deliquescence.

In the experiment, the percentage of water in a hydrate will be determined using the
relationship given above. The formula of the hydrate may then be determined by calculating the
moles of anhydrous salt and the moles of water of hydration. Their molar ratio is then
determined. This relates to the general formula of the hydrate. For example, suppose that the
hydrate in question was sodium sulfate decahydrate, Na2SO410H2O. The original sample of the
hydrate weighed 320.0mg. After heating the anhydrous salt weighed 141.3 mg. The mass of
water lost would therefore be 320.0 -141.3 = 178.7 mg. The number of moles of water lost
would be

23
 0.1787
= = = 9.918 10
18.016

The number of moles of sodium sulfate would be

ℎ 0.1413
= = = 9.954 10
141.96

From the ratio of the number of moles,

9.918 10
= = 9.964
9.954 10
it is readily seen that there are 10 mole of water associated with each mole of the sodium
sulfate.
Chemicals and apparatuses
Unknown hydrate, balance, aluminum foil, beaker, measuring cylinder, desiccators, tongs,
crucible, distilled water.

Procedures
Part A: Visual Inspection of a Series of Hydrates and Anhydrous Salts
Inspect the group of hydrates and anhydrous salts provided by your instructor.
Comment on the Data Sheet on their physical appearance and color. Which show deliquescent
properties? Which might find use as desiccants? Do any exhibit efflorescence?

Part B: Formula of a Hydrate by Gravimetric Analysis

The analysis should be done in triplicate. Mark three crucibles A, B, and C in pencil.
While one crucible is cooling, the next can be heated. Attach an iron ring to a ring stand. Place a
clay triangle on the ring. Put the 1st crucible in the clay triangle and heat to redness with an
intense flame using a Bunsen burner for approximately 5 minutes.

Reminder: Use forceps or tongs to handle the crucibles,

never your fingers!

Place the crucible in a small beaker and then place the beaker in the desiccators. While the first
crucible is cooling, start heating crucible 2. Allow them to cool to room temperature. Mass of

24
each crucible. Repeat heating, cooling and weighing until a constant mass for each of the
crucibles is obtained.

Add 1 gram of sample of the unknown hydrate to each of the crucibles and weigh. Be sure to
record all of the decimal places on the balance. Place a small piece of Aluminum foil (do not
cover the crucible entirely) on each crucible to serve as a cover. Place one crucible (with cover)
in the triangle. Heat the sample gently at first and then increase the heat moderately. Do not
permit the crucible to become red hot. Continue the heating for approximately 2 to 3 minutes.
Remove the crucible and place it in a beaker. Transfer the beaker and crucible to a desiccators
to cool to room temperature. Process the second sample in the same manner, followed by the
third. Weigh each crucible, without the cover and record the results on the Data sheet.

To make sure all of the water of hydration has been removed, reheat each sample, cool as
before and reweigh. Repeat this process until each sample achieves a constant mass. Note: the
masses for the samples (A, B and C) may be different from each other but should be consistent
within the sample. Record your results on the Data Sheet.

Determination of the Empirical Formula of a Hydrate

Data Sheet

Part A: Visual Inspection of a Series of Hydrates and Anhydrous Salts. Write concise
statements of your observations for each of the samples. CuSO4, CuSO45H2O, Na2SO4,
Na2SO410H2O, CaCl2, CoCl26H2O, NaOH, CoCl2.

25
Part B: Gravimetric Analysis of an Unknown Hydrate

Crucible Crucible B Crucible C

A
1. Constant mass of crucible
1st heating (g)
2nd heating (g)
3rd heating (g) if necessary

2. Mass of Crucible and hydrated salt

(g)

3. Constant mass of crucible and

anhydrous salt
1st heating (g)
2nd Heating (g)
3rd heating (g) if necessary

4. Calculations
Mass of the hydrated salt (g)
Mass of the anhydrous salt (g)
Mass of water lost (g)
Percentage of water in hydrate
Average of % water in the unknown

Formula of the anhydrous salt

Moles of water lost
Average moles of water lost
Moles of anhydrous salt obtained
Average moles of anhydrous salt
Mole ratio of water to anhydrous salt
(based on average values)

Formula of the unknown Hydrate

26
Post lab. Questions

1.) You are given a sample of an unknown hydrate of CaSO4. The average values of the data
collected and recoded in your laboratory notebook where as follows:

Mass of empty crucible 31.138 g

Mass of crucible and sample before heating 31.457 g

Mass of crucible and sample after heating 31.436 g

Calculate the formula of the CaSO4 hydrate.

2.) Sodium hydroxide pellets are very hygroscopic. Describe what happens chemically when a
cover is left off a bottle of NaOH pellets. (Hint: Carbon dioxide reacts with NaOH.)

3.) A 120.0 mg sample of a lead acetate hydrate, Pb (C2H3O2)2xH2O was heated to drive off the
waters of hydration. The cooled residue weighed 103.0 mg. Calculate the value of x in the
acetate formula.

27
 PART 3: CHROMATOGRAPHIC
SEPARATION
Chromatography is a technique for separation, identification, and determination of the chemical
the components, or solutes, of a mixture on the basis of the relative amounts of each solute
distributed between a moving fluid stream, called the mobile phase, and a contiguous stationary
phase. The mobile phase may be either a liquid or a gas, while the stationary phase is either
a solid or a liquid. It is an analytical technique commonly used for separating a mixture of
chemical substances into its individual components, so that the individual components can be
thoroughly analyzed. There are many types of chromatography such as paper chromatography,
thin layer chromatography, column chromatography, gas chromatography, HPLC
chromatography, ion-exchange chromatography, affinity chromatography, size- exclusion
chromatography but all of these employ the same basic principles i.e. The separation is based
on differential partitioning between the mobile and stationary phases. Mobile phase: it is a gas
or liquid moves through a definite direction, mean that moves through the chromatography
column or plate (stationary phase) while the sample interacts with the stationary phase and is
separated. Stationary phase: it is either liquid or solid the so called immobilized phase ,
immobilized on the support particles or on the inner wall of the column tubing.

28
EXPERIMENT 1: PAPER CHROMATOGRAPHY OF DYES

Objective: to get acquainted with different techniques of chromatography. To separate organic

dyes by paper chromatography using different mobile phases

Theory

Chromatography is used to separate mixtures of substances into their components. All forms of
chromatography work on the same principle. They all have a stationary phase (a solid or a
liquid supported on a solid) and a mobile phase (a liquid or a gas). The mobile phase flows
through the stationary phase and carries the components of the mixture with it. Different
components travel at different rates. In paper chromatography, the stationary phase is a very
uniform absorbent paper. The mobile phase is a suitable liquid solvent or mixture of solvents.
Paper chromatography is a technique that involves placing a small dot or line of sample solution
onto a strip of chromatography paper. The paper is placed in a container with a shallow layer
of solvent and sealed. As the solvent rises through the paper, it meets the sample mixture, which
starts to travel up the paper with the solvent. This paper is made of cellulose, a polar substance,
and the compounds within the mixture travel farther if they are non-polar. More polar
substances bond with the cellulose paper more quickly, and therefore do not travel as far, in
other words In paper chromatography, the sample mixture is applied to a piece of filter paper,
the edge of the paper is immersed in a solvent, and the solvent moves up the paper by capillary
action. Components of the mixture are carried along with the solvent up the paper to varying
degrees, depending on the compound’s preference to be adsorbed onto the paper versus being
carried along with the solvent. The paper is composed of cellulose to which polar water
molecules are adsorbed, while the solvent is less polar, usually consisting of a mixture of water
and an organic liquid. The paper is called the stationary phase while the solvent is referred to as
the mobile phase. In order to obtain a measure of the extent of movement of a component in a
paper chromatography experiment, we can calculate an “Rf value” for each separated
component in the developed chromatogram. An Rf value is a number that is defined as: distance
traveled by component from application point or The distance travelled relative to the solvent
expressed as:

29
 Fig.1. set up of paper chromatography

Chemicals and apparatuses

Developing chamber, measuring cylinder, capillary tube, chromatography paper, dyes

(methyl red, methyl violet, Congo red, bromocresol green ), mobile phase( acetone,
ethyl acetate, acetone: ethyl acetate (50:50).

Procedures

 Fill the tanks with the different solvents. Pour 50ml of the solvent on the bottom
the tank and replace the lid. The tanks must always be covered to avoid
evaporation and to allow the atmosphere to become saturated.
 Cut a sheet of paper of the format 15x15cm for applying 5 spots. Draw a pencil
line 2.5 cm from one edge across the paper and mark the origins for the spots
with pencil. The first is 3.5cm from the side. Then a space of 2cm between 2
successive spots.
 Spot the test solution using capillary tube. The spots should not be greater than
7mm in diameter.

30
  After the spots have dried roll the paper to form a cylinder with the starting line
at one end. Fasten the edge together and put the cylinder into the tank. Make sure
it is standing freely and not touching the glass walls.
 Close the lid and allow the chromatogram to run.
 After the rise of the mobile phase of about 12cm, remove the chromatogram,
mark the solvent front, open the cylinder and dry the paper. Mark the spots and
measure the distance of each spots.

Post lab. Questions

 Calculate the Rf values for all dyes investigated in the ascending chromatography
and compare the results obtained with different mobile phases and discuss the
results with respect to the polarity of the mobile phases.

EXPERIMENT 2: THIN LAYER CHROMATOGRAPHY

Objective: to separate organic dyes by TLC using different mobile phases and to see the
influence of solvent polarity on the Rf values of the dyes.

Theory

Thin-layer chromatography (TLC) is a chromatography technique used to separate non-volatile

mixtures. Thin-layer chromatography is performed on a sheet of glass, plastic, or aluminum
foil, which is coated with a thin layer of adsorbent material, usually silica gel, aluminum
oxide (alumina), or cellulose. This layer of adsorbent is known as the stationary phase.

After the sample has been applied on the plate, a solvent or solvent mixture (known as
the mobile phase) is drawn up the plate via capillary action. Because different analytes ascend
the TLC plate at different rates, separation is achieved. The mobile phase has different
properties from the stationary phase. For example, with silica gel, a very polar substance, non-
polar mobile phases such as heptanes are used. The mobile phase may be a mixture, allowing
chemists to fine-tune the bulk properties of the mobile phase.

After the experiment, the spots are visualized. Often this can be done simply by
projecting ultraviolet light onto the sheet; the sheets are treated with a phosphor, and dark spots
appear on the sheet where compounds absorb the light impinging on a certain area. Chemical

31
processes can also be used to visualize spots; anisaldehyde, for example, forms colored adducts
with many compounds, and sulfuric acid will char most organic compounds, leaving a dark spot
on the sheet.

To quantify the results, the distance traveled by the substance being considered is divided by the
total distance traveled by the mobile phase. (The mobile phase must not be allowed to reach the
end of the stationary phase.) This ratio is called the retention factor or Rf. In general, a

substance whose structure resembles the stationary phase will have low R f, while one that has a

similar structure to the mobile phase will have high retention factor. Retention factors are
characteristic, but will change depending on the exact condition of the mobile and stationary
phase. For this reason, chemists usually apply a sample of a known compound to the sheet
before running the experiment.

Thin-layer chromatography can be used to monitor the progress of a reaction, identify

compounds present in a given mixture, and determine the purity of a substance. Instrumental set
up and its principle is similar with paper chromatography.

Chemicals and apparatuses

Pre-coated TLC plates, developing chamber capillary tubes, acid dyes(methyl red, congo red,
bromocresol green and mixtures). Mobile phases 1. 18ml acetone and 12ml of hexane, 2.14ml
acetone and 14ml ethyl acetate, 3. 14ml acetone and 14 ml ethanol.

Procedures

 Take three developing chambers

 Prepare the three mobile phases by mixing the solvents in the given volume ratio by
means of graduated cylinders follows

Mp1. 18ml acetone and 12ml of hexane

Mp2. 14ml acetone and 14ml ethyl acetate

Mp3. 14ml acetone and 14ml ethanol

 Fill the three chromatographic tanks with each of the mobile phases to about 0.5cm high
and close the lid for vapor saturation.

 Prepare three aluminum TLC plates by marking lightly with pencil(not ink) a line 2cm
from the bottom and 1cm from the top.

 From the bottom line, mark four small pencil dots to make where the dye spots will be
applied.
32
  Above the top line write the letters corresponding to the four dots of the dyes.

 Using a separate capillary tube for each dye, apply a single drop of each dye to the
match in pencil dot on the bottom line of each plate

 Place the three plates in the three developing chambers containing the solvents and close
the lid

 When the solvent on each plate has risen to the upper pencil line , remove the plate from
its chamber and mark the exact position of the solvent front before it evaporates.

 Mark the plate with the identity of the solvent

 The sample of the plates, draw a pencil line on each plate 2cm above the edge of the
plate and apply the samples as small spots

 Place the plates in the tanks and close them. Wait until the solvent front is about 2cm
below the upper edge of the plate. Then remove the plates from from the tank, dry the
lower edge of the plate from excess solvent using filter paper, and mark the solvent front
with a pencil.

 Dry the plates, outline the spots, and measure the distance between center of the spot
and origin.

 For dye on each plate, determine the Rf value and record the value.

Post lab. Questions

1. Explain the differences and similarities between paper and thin layer chromatography.

2. What detection methods are used when spots are colorless?

3. Why it is necessary to close the tanks during the development of the chromatograms?

4. Which solvent mixture gave the most complete resolution of the three dyes? Which solvent
mixture gave the poorest resolution of the three dyes?

33
EXPERIMENT 3: COLUMN CHROMATOGRAPHY

Objective: to separate and identify plant pigments by using column chromatography

Theory

Column chromatography is one of the most useful methods for the separation and purification
of both solids and liquids. This is a solid - liquid technique in which the stationary phase is a
solid & mobile phase is a liquid. The principle of column chromatography is based on
differential adsorption of substance by the adsorbent.
The usual adsorbents employed in column chromatography are silica, alumina, calcium
carbonate, calcium phosphate, magnesia, starch, etc., selection of solvent is based on the nature
of both the solvent and the adsorbent. The rate at which the components of a mixture are
separated depends on the activity of the adsorbent and polarity of the solvent. If the activity of
the adsorbent is very high and polarity of the solvent is very low, then the separation is very
slow but gives a good separation. On the other hand, if the activity of adsorbent is low and
polarity of the solvent is high the separation is rapid but gives only a poor separation, i.e. the
components separated are not 100% pure.

The adsorbent is made into slurry with a suitable liquid and placed in a cylindrical tube that is
plugged at the bottom by a piece of glass wool or porous disc. The mixture to be separated is
dissolved in a suitable solvent and introduced at the top of the column and is allowed to pass
through the column. As the mixture moves down through the column, the components are
adsorbed at different regions depending on their ability for adsorption. The component with
greater adsorption power will be adsorbed at the top and the other will be adsorbed at the
bottom. The different components can be desorbed and collected separately by adding more
solvent at the top and this process is known as elution. That is, the process of dissolving out of
the components from the adsorbent is called elution and the solvent is called is called eluent.
The weakly adsorbed component will be eluted more rapidly than the other. The different
fractions are collected separately. Distillation or evaporation of the solvent from the different
fractions gives the pure components.
The main advantage of column chromatography is the relatively low cost and disposability of
the stationary phase used in the process. The latter prevents cross-contamination and stationary
phase degradation due to recycling. The classical preparative chromatography column is a glass

34
tube with a diameter from 5 mm to 50 mm and a height of 5 cm to 1 m with a tap and some kind
of a filter (a glass frit or glass wool plug – to prevent the loss of the stationary phase) at the
bottom. The stationary phase or adsorbent in column chromatography is a solid. The most
common stationary phase for column chromatography is silica gel, the next most common
being alumina and the mobile phase or eluent is either a pure solvent or a mixture of different
solvents.

Fig 1 Column chromatography setup

Chemicals and apparatus

Beaker 100ml, beaker 600ml, Erlenmeyer flask 100ml, pipette, pipette bulb, mortar and pestle,
glass rod, cork ring, swan- neck lamp, silica gel, petroleum ether, acetone, sodium chloride,
sodium sulfate, fresh leaves, glass chromatography column, separation funnel, reparatory
funnel, measuring cylinder.

Procedures

Extraction of the leaf pigments

Using a pestle, fresh leaves are grinded in mortar containing 22ml of acetone, 3ml of petroleum
ether, and spatula tip-full of calcium carbonate. The pigment extract is filtered. The filtrate is
poured into a separation funnel and it is mixed with 20ml of petroleum ether and 20ml of 10%
aqueous NaCl solution. The separating funnel is shaken carefully. When the layers have
separated the lower layer is allowed to drain in to a beaker. The lower phase is thrown away.
The upper layer is washed 3-4 times with 5ml of distilled water. After wards the extract is
decanted into an Erlenmeyer flask. Eluting solvent (mobile phase) : mixture of petroleum ether
and acetone(7:3) and silica gel slurry: using a beaker of an appropriate size, slurry of silica gel,
and eluting solvent is prepared.

35
 Packing a (silica gel) column:

1. Use a piece of wire to add a plug of cotton to the bottom of the column. There should be enough
cotton that the sand and silica will not fall out of the column. However, too much cotton or cotton
packed too tightly will prevent the eluent from dripping at an acceptable rate.
2. Clamp the column to a ring stand and add enough sand to fill the curved portion of the column.
3. Place a pinch clamp on the tubing, then fill the column 1/4 to 1/3 full with the initial eluent. (The
composition of eluent is often changed as the separation proceeds.)
4. Prepare slurry of silica in the initial eluent by pouring dry silica into a beaker of eluent. (Add a volume
of silica gel, such as 20 ml, to approximately double the volume of eluent, 40 ml).
CAUTION: keep the dry silica in your hood and be careful not to inhale the lightweight substance.

5. Quickly but carefully pour the slurry into the column. Stir and pour immediately to maximize the
amount of silica that goes into the column instead of remaining behind in the beaker. You may find a
clean spatula or glass rod helpful in transferring the silica.

6. Remove the pinch clamp to allow solvent to drip into a clean flask. Tap on the side of the column with
a rubber stopper or tubing to help the silica settle uniformly.
7. Use a Pasteur pipette to rinse any silica that is sticking to the sides of the column. Allow the silica to
settle while eluent continues to drip into the flask.
8. Once the silica has settled, carefully add sand to the top of the column. Sand is heavier than silica. If
the silica has not settled, the sand may sink into the silica instead of forming a layer on top of it. (You
may need to rinse down sand that sticks to the side of the column.

Loading a sample onto the column:

1. Drain eluent from the column until no solvent remains above the surface of the sand.
2. Using a long Pasteur pipette, carefully add your sample to the column.
3. Drain eluent from the column until no sample remains above the surface of the sand.
4. Use ~ 1 ml of eluent to rinse your container and pipette. Add this milliliter of sample to the sand.
Drain eluent from the column until no liquid remains above the surface of the sand.
5. Repeat step 12 two or three times to completely transfer your sample onto the silica gel. If you do
not do and repeat step 12, your sample will remain in the sand instead of on the silica. Sample
remaining in the sand will dissolve in the eluent that you add in step 14, ruining the possibility of
good separation of components.

36
 Eluting the sample:

1. Once you have rinsed your sample onto the silica, carefully add eluent to the top of the column. To
avoid disturbing the top of the column, it's a good idea to carefully pipet an inch or two of solvent
onto the column instead of pouring solvent directly onto the sand.
2. Add more eluent as necessary. The eluent collected prior to the elution of sample can be recycled.
The composition of the eluent can be changed as the column progresses. If the eluent composition is
to be changed, ALWAYS start with least polar solvent/mixture and change to the more polar
solvent/mixture.

Experimental procedures

Adding the Sample

Once the packing is complete, 20ml of the sample/leaf extract can be loaded directly to the top
of the column. Normally, a minimum amount of a polar solvent, 5-10 drops, is used to dissolve
the mixture. The solution is then carefully added to the top of the column using a pipet without
disrupting the flat top surface of the column. A thin horizontal band of sample/leaf extract is
best for an optimal separation. After the sample/leaf extract is loaded, a small layer of white
sand is added to the top of the column. This will help to keep the top of the column level when
adding solvent eluent. Once the mixture is added and the protective layer of sand is in place,
continuously add the solvent eluent while collecting small fractions at the bottom of the
column. Using a pipette to add the first bit of solvent on top of the packing, sample, and sand
will minimize disturbance of the column and diluting the sample. Collecting small fractions (1-
3 ml) is important to the success of your column separation.

Post lab. Questions

1. Discuss briefly the differences and similarities between thin layer and column
chromatographic techniques.

2. What type of experimental error could influence the outcome of the separation?

37
 PART 4: SELECTED EXPERIMENTS ON
ELECTRO ANALYTICAL METHODS
Electro analytical methods are a class of techniques in analytical chemistry which study
an analyte by measuring the potential (volts) and/or current (amperes) in an electrochemical
cell containing the analyte.[1][2][3][4] These methods can be broken down into several categories
depending on which aspects of the cell are controlled and which are measured. The main
categories are potentiometry (the difference in electrode potentials is
measured), coulometry (the cell's current is measured over time), and voltammetry (the cell's
current is measured while actively altering the cell's potential), conductivity measurement.
Among those categories we will see conductivity measurement.

EXPERIMENT 1: MOLAR CONDUCTIVITIES OF AQUEOUS ELECTROLYTES

Objective: To determine the conductivity of various acids and the dissociation constant, Ka for
acetic acid

Theory

We all know that the solutions of electrolytes conduct an electric current by the motion of the
cations towards the cathode, and anions towards the anode. Metallic conductors are known to
obey Ohm’s Law:

I = ΔΦ /R (1)

Where, I is the current in amperes, ΔΦ is the difference in the electric potential between the
terminals of the conductor in volts, and R is the resistance in ohms. R depends upon the
dimensions of the conductor:

R = ρl/A (2)

Where l is length, A is the cross-sectional area and ρ is the special resistance, i.e. resistivity, in
Ωm. Reciprocal resistance, 1/R, is known as conductance, and reciprocal resistivity, 1/ρ, as
conductivity. Conductivity is given by the symbol kappa, κ and has units Ω -1m -1 or Sm-1 (S =
Ω -1). Hence,

κ = l/ RA (3)

Conductivity depends upon the number of charge carriers (i.e. ions) present in the solution. It is
therefore, reasonable to express it in terms of molar conductivity:

Λ m = κ/ c (4)

38
where c is the molar concentration of the electrolyte. We can classify electrolytes as a) strong
electrolytes, and b) weak electrolytes. The strong electrolytes have high molar conductivities,
and their conductivities change very little if the solution is diluted down. In the case of weak
electrolytes, however, there is little conductivity in concentrated solutions. The conductivity
increases upon increasing dilution. The value of Λm extrapolated to zero concentration (i.e.
theoretical infinite dilution) is known as Λm0, the molar conductivity at infinite dilution when
ion interaction is assumed to be zero. It is also known as the limiting molar conductivity.
Kohlrausch’s Law: Λm =Λmo− Κ c1/2 (5)

Where, K is an experimental constant. One can obtain the value of Λm0 from the intercept of
1/2
the slope of the Λm versus c plot. Since each ion is assumed to be migrating independently,
the value of Λm0 can also be obtained from the addition of individual conductivities of ions:

Λmo= Λ+o + Λ-o (6)

Or Λmo = ν+ λ+ + ν- λ- (7)

where ν+ and ν- are the numbers of cations and anions per formula unit of the electrolyte, and
λ+ and λ- are the limiting ionic conductivities. For a dilute, weak electrolyte

Λm= αΛmo (8)

Where α is the degree of ionization as in the dissociation for acetic acid HAc given below:

HAc + H2O AC- + H3O + (9)

(1-α)c αc αc

where c is the molar concentration. For this acid dissociation constant Ka can be given by

Ka = [Ac- ][ H 3O+]/ HAc = α2 c/ 1- α (10)

Chemicals and apparatuses

Conduct meter, beaker, deionized (distilled) water, 0.02 M KCl solution, 0.02 M KAc, 0.02 M
HCl and 0.05 M HAc, measuring cylinder.

39
Procedures

1. Read the cell constant, x, for the cell used in your experiments from the conductometer.

x = кR (11)

Where, к is the conductivity and R is the resistance, as previously.

2. Calibrate the conductometer by immersing the conductivity cell into a beaker, filled with
deionized (distilled) water. Set the knob to red line and adjust the scale.

3. Rinse the conductivity cell with 0.02 M KCl, then fill a beaker with 100 ml of this solution
and immerse the conductivity cell.

4. Keep the conductivity cell steady in the solution and adjust the knob to “conductance”. Read
and record the value from the upper scale (in units of μmhos).

5. Take 25 ml of the 0.02 M KCl solution into a beaker and add 75 ml distilled water. Repeat
steps 2-4 with this solution.

6. Prepare 0.02 M KCl solutions at 1/16 and 1/64 dilutions in the same way explained in step 5
and repeat steps 2-4 with these solutions. Serial dilutions are strongly encouraged.

7. Repeat steps 5-6 with solutions of 0.02 M KAc, 0.02 M HCl and 0.05 M HAc.

Calculations

1. Determine the Λm values using equation (4). Make sure that you obtain the correct units.
(Λm = Sm2 /mol)

2. Tabulate the R, κ, and Λm values for all the solutions.

3. Using equation (5), plot Λm versus c 1/2 for each solution to determine the Λm0 values.

4. using the Kohlrausch Law, determine the Λm0 for acetic acid:

Λm0 (HAc) = Λm0 (HCl) + Λm0(KAc) - Λm0(KCl) (12)

5. Using equation (8), determine α for each HAc concentration used, and calculate Ka (again for
each case) using equation (10).

6. Determine the average Ka value for all the experiments.

7. Alternatively, determine Ka by extrapolation to “zero concentration” on a Ka versus (αc)1/2

plot.

8. Compare the results obtained in steps 6 and 7. Repeat steps 5-6 with solutions of 0.02 M
KAc, 0.02 M HCl and 0.05 M HAc.

40
Post lab. Questions

1. Determine which solution has a higher conductivity, 0.1 M HCl or 0.1 M HAc. Explain
what creates the higher conductivity.

2. Discuss about strong electrolytes, weak electrolytes and non electrolytes and compare in
terms of conductivity.

EXPERIMENT 2: DETERMINING THE PH OF VARIOUS SUBSTANCES

Objective: The purpose of this experiment is to measure the pH of various substances using
three unique methods.

Theory

The degree to which a substance is acidic or basic is referred to as its pH. The pH scale is
measured from a scale of 0 to 14. A solution having a pH of 7 is considered to be neutral, that is
neither acidic nor basic. A solution with a pH less than 7 is considered acidic and a pH greater
than 7 is considered basic. However there are varying degrees as to how acidic or basic a
solution may be. If a solution has a pH between 0 and 2, the solution is considered very acidic
while a solution between 2 and 5 is considered moderately acidic. A solution having a pH
between 5 and 7 is slightly acidic. If a solution has a pH between 7 and 9, then the solution is
considered slightly basic and a solution having a pH between 9 and 11 is considered moderately
basic. A solution with a pH between 11 and 14 is very basic. You will need this information
when deciding how acidic or basic a substance is after determining the pH.

There are many ways to determine the pH of a substance or solution. The methods you will use
will be using a chemical indicator, pH paper, and a pH meter. The simpler methods are the first
two mentioned previously. When you place a chemical indicator (chemical dye) into the
solution you wish to analyze, the indicator will change to a specific color, depending on the pH
of the solution. When using pH paper, you can use either pH paper that just indicates whether a
solution is acidic or basic, or you can use universal pH paper which gives you the entire pH
scale as an assortment of colors. That means you cannot only determine whether your solution
is acidic or basic, you can also determine how acidic or basic you solution is depending on the
color change of the pH paper. The methods you will use today is the last method using a pH
meter, which can measure the pH of a solution electronically. You must standardize the pH
meter before each measurement with solutions where the pH is known. Once the pH meter is
41
standardized, you can measure the pH of an unknown substance. Your instructor will teach you
how to properly use a pH meter before you begin measuring the pH of any unknown
substances.

Chemicals and apparatuses

PH meter, 0.1 M sodium hydroxide (NaOH), 0.1 M, Hydrochloric acid (HCl), pH buffers,
distilled water

Procedures

In this experiment the solutions you will analyze are 0.1 M sodium hydroxide (NaOH), 0.1 M

Hydrochloric acid (HCl), Record all of your results in the table on your results page. Procedure
(continued)

1. Obtain beakers and label each with the name of one of the substances you are going to
analyze.

2. Pour 10-20 ml of each solution in the properly labeled beaker depending on the size of your
beaker.

3. Obtain approximately 20mL of pH 7 buffer and place in a beaker labeled buffer. You will use
this solution to calibrate your pH meter. Ask your instructor for instructions on how to properly
use a pH meter.

4. When you have learned to properly use the pH meter, calibrate then rinse the electrode. Place
the electrode in one of the solutions being analyzed and wait for the digital reading on the meter
that will tell you the pH of that solution. When you have recorded this value, rinse the electrode,
calibrate the pH meter and rinse the electrode again. Place the electrode in another solution you
wish to analyze.

5. Repeat step 4 until you have analyzed all of your solutions.

6. When you are finished, properly dispose of the solutions

Record all results

42
Post Lab Questions

1. What is the advantage of calibrating pH meter with buffer solutions before measuring
the sample and why to rinse the electrodes?
2. Calculate the pH, pOH, [H+] and [OH-] of 0.02 mol/L H2SO4?

PART 5: EXPERIMENTS ON
SPECTROSCOPY
Spectroscopy is the study of the interaction between matter and electromagnetic radiation. It is a
scientific measurement technique; It measures light that is emitted, absorbed, or scattered by
materials and can be used to study, identify and quantify those materials. When a beam of
electromagnetic radiation passes through a sample, the photons interact with the sample. They
may be absorbed, reflected, refracted, etc. Absorbed radiation affects the electrons and chemical
bonds in a sample. In some cases, the absorbed radiation leads to the emission of lower energy
photons. Spectroscopy looks at how the incident radiation affects the sample. Emitted and
absorbed spectra can be used to gain information about the material. Because the interaction
depends on the wavelength of radiation, there are many different types of spectroscopy such as
uv-visible spectroscopy, infra-red spectroscopy, nuclear magnetic resonance spectroscopy,
Raman spectroscopy, mass spectroscopy. Also we can classify spectroscopic methods according
to the region of the spectroscopic spectrum involved in the measurement. The regions of the
spectrum that have been used include gamma ray, X-ray, ultraviolet, visible, infrared,
microwave, and radio frequency. But for the sake of simplicity we will focus only on
ultraviolet-visible spectroscopy.

43
EXPERIMENT 1: UV/VIS SPECTROSCOPY AND SPECTROPHOTOMETRY:
SPECTROPHOTOMETRIC ANALYSIS OF POTASSIUM PERMANGANATE
SOLUTIONS.
Objectives: After completing this experiment, the student should be able to:

1. Prepare standard solutions of potassium permanganate.

2. Construct calibration curve based on Beer’s Law.

3. Use Beer’s Law to determine molar absorptivity.

4. Explain the fundamental principal behind Spectrophotometric analysis

Theory

Most analytical methods require calibration (a process that relates the measured analytical
signal to the concentration of analyte, the substance to be analyzed). The three most
common analysis methods include the preparation and use of a calibration curve the
standard addition method, and the internal standard method. In this experiment we will
use Spectrophotometry to prepare a calibration curve for the quantitative analysis of
KMnO4.

SPECTROPHOTOMETRY
Spectrophotometry is a technique that uses the absorbance of light by an analyte (the
substance to be analyzed) at a certain wavelength to determine the analyte concentration. UV/
VIS(ultra violet/visible) Spectrophotometry uses light in UV and visible part of the
electromagnetic spectrum. Light of this wavelength is able to effect the excitation of electrons
in the atomic or molecular ground state to higher energy levels, giving rise to an absorbance at

wavelengths specific to each molecule. The complex formed between ASA and Fe3+is
intensely violet colored, and therefore can be determined by Spectrophotometry using the
visible part of the electromagnetic spectrum. The human eye is able to “see’ light the

wavelength range 400-700nm (nanometeror10−9m). To the human eye, wavelength appears

as color, as shown in the following table.
Table: Correlation between wavelength, color, and complementary color
Wavelength, nm Colour Complementary colour
400-430 Violet Yellow-green
430-490 Blue Yellow
490-550 Green Purple
550-590 Yellow Blue
590-630 Orange Green-blue
630-700 Red Blue-green

44
 When we see an object as purple, in fact it absorbs light in the “green” region of the spectrum:
only blue and red wavelengths reach the eye, which we experience, as the purple color. The

ASA-Fe3+ complex formed in the analysis of aspirin absorbs wavelengths in the blue, green
and yellow regions of the spectrum, as shown in figure1. Only violet and red light are
“transmitted”. As a result, the human eye sees the transmitted light as a mixture of violet and
red, which we experience as a deep purple.

Figure 2: The absorption spectrum of the Fe3+- salicylate

complex.
InFig.2, note that %Transmittance T is plotted on the left (linear scale), Absorbance A on the
right (logarithmic scale).The absorbance maximum is atapproximately510 nm (λmax).

The spectrophotometer takes the place of the human eye by accurately measuring the intensity
of the light transmitted at each wavelength. For Spectrophotometry in the visible region the
light source is a common tungsten light bulb emitting “white” light. The light is collimated
and focused on an entrance slit, and then falls on a monochromator, that separates the white
light in its constituent wavelengths. The monochromator can be a glass prism, but in modern
instruments it will be a grating. From the monochromator the light is sent through the sample,
and finally reaches a photocell that measures the intensity of the light at each specific
wavelength.

Spectrophotometers have many different designs. In the simplest instrument, for each
wavelength λ set by the monochromator, the user first inserts a blank solution in place of the
sample. The photo cell then records the intensity at that wavelength. This intensity is called Io
(λ).Next the blank solution is replaced by the sample solution, the photometer measures the
new intensity called I (λ). The Transmittance, T is then displayed on the screen or
45
spectrophotometer output:

T (λ) = I (λ)/IO (λ) (1)

Often the Transmittance is expressed as a percentage

%T(λ)=I(λ)/IO()x100% (2)

This procedure is then repeated for a number of

wavelengths.

More sophisticated instruments “scan” the spectrum over the required wavelength
range automatically, and record the transmittance as a function of wavelength. In a “double
beam” scanning spectrophotometer, part of the light is reflected to a separate blank cell, and
the intensities I and I0 are measured simultaneously at each wavelength, and automatically
compared to yield a direct output of T vs λ. Another modern form of the UV/VIS
spectrophotometer is the “Diode Array” spectrophotometer. In this instrument the photocell,
which measures light intensity at one wavelength at a time, is replaced by a CCD detector
similar to the detector in your digital camera, and the instrument can record the spectrum over
the full wavelength range (typically 200−700 nm) within one second!
In each fractional layer of the sample the intensity of the incoming light will decrease
proportionally to the concentration of the analyte. As a result, the Transmittance T decreases
exponentially with increasing path length l . This is expressed in the Beer Lambert law, which
states:

-log I/IO = ε .l. C (3)

ε is called the “Molar Absorptivity” of the compound. It is a function of wavelength specific

for each molecule. With the path length l normally given in cm, and C in Molarity units,
mol/L ε has the units [Link]−1cm−1. If alternatively C is in mol/ml, then ε will have the unit’s
cm2mol−1.

The Absorbance, A, is defined as

A = -log I/IO (4)

46
Thus, the Beer-Lambert law, more commonly called Beer’s law, can be written as:

A = ε .l. C (5)
Finally, the relation between A and T is given by:

A = -log T (6)

Note that A is a dimensionless quantity. Because ; A is directly proportional to the

analyte concentration, it is more often used than T or %T. Most spectrometers can record
either A, T, or %T for a given wavelength.

The calibration curve method

To use the calibration curve technique, several standards containing exactly known
concentrations of the analyte are introduced into instrument, and the instrumental response is
recorded. Ordinarily this response is corrected for instrument output obtained with a blank.
Ideally, the blank contains all of the components of the original sample except for the
analyte. The resulting data are then plotted to give a graph of corrected instrument response
versus analyte concentration, and this graph in turn can be used to find the concentration of
an unknown.

Solutions of analyte may absorb light of different wavelengths. When the analyte to our eyes
is a coloured solution, this means that out of the white light visible to the eye certain
wavelengths are absorbed by the analyte molecules, letting the other colours of the white light
pass through, giving rise to the characteristic colour of the analyte in solution. For instance,
the analyte in this experiment, potassium permanganate or KMnO4 absorbs light in the green
region of the visible spectrum, letting red light and blue light pass through. The mixture of
blue and red is perceived by the eye as the
typical purple colour of KMnO4.

A spectrum shows how much light is absorbed for a range of wavelengths. In UV/VIS
spectrophotometry we plot the Absorbance A of a solution (A is a measure of how much
light is absorbed, see below), against the wavelength of the light reaching the solution, λ.
This is called the absorption spectrum.

47
 The higher the analyte concentration, the more light at a certain wavelength will be
absorbed. The relation between Absorbance A and analyte concentration C is given in
equation 5:

In a UV/VIS spectrophotometer a light source emits light at all wavelengths of the UV and
visible spectrum. Via a system of mirrors the light is guided to a monochromator that selects
light of only a certain wavelength. After the monochromator, the light passes through the
analyte sample solution held in a cuvette in the sample holder, and finally reaches the detector
(a photocell), where the intensity of the light reaching it is measured, and recorded as the
Absorbance A or the Transmission T (or %T). The relation between A, T and %T is given in
the spectrophotometry section of Experiment 10, but in this experiment we will only use the
Absorbance at different wavelengths, A(λ).

In a single beam spectrophotometer (as used in this experiment), we first place a cuvette filled
with solvent only (water in this case), to measure the intensity of the light source at a certain
wavelength. Next, we place a cuvette filled with sample solution in the cell holder. The
instrument will record light intensity and absorbance relative to the light intensity passes
through solvent (water) alone. This must be repeated at different wavelengths to obtain the
spectrum of the dissolved solute (A (λ) vs. λ).

From the spectrum we find the wavelength with the highest absorbance, the wavelength of the
absorption peak, λmax. At this wavelength the spectrophotometric method is most sensitive
for the analyte. Next, we determine the absorbance A at λmax for a number of standard
solutions of different concentration, always starting with the lowest concentration. From these
absorbance values, we plot a calibration curve for the analyte (KMnO4 in our case), which we
can then use to determine the concentration of an analyte solution of unknown concentration.

Materials and Equipment

UV/VIS spectrophotometer and polystyrene cuvettes, 2 100 mL volumetric flask, 1000 μL

micropipette, 100 μL pipet, Hot plate, solid KMnO4, 1 10 mL graduated cylinders, 6 10
mL volumetric flasks, 2 125 mL Erlenmeyer flasks or 150 mL beakers
Procedure
Preparing the stock solution and four standard solutions
1. Preparation of 100 mL of a stock standard solution of 0.008M KMnO4: Accurately weigh
126 mg solid KMnO4. Transfer quantitatively to a 100 mL volumetric flask and fill to the

48
mark with water. This is the stock solution.

2. Prepare four standards in 10.o mL volumetric flask with concentrations of 0.00008 M

(solution #1),
0.00016 M (solution #2), 0.0004 M (solution #3) and 0.0008 M (solution #4) by diluting
the stock solution prepared in Step 1 as following.

For the 0.00008, 0.00016 and 0.0004 M standards use the 100 μL micropipets (100 μL = 0.1
mL; calculate how many 100 μL samples stock solution are needed in each case) to make 10
mL standard [Link] the 0.0008 M standard use the 1 mL (1000 μL) micropipet, Mark
the four 10 mL flasks with standard solutions #1-4 (lowest concentration #1 = 0.00008 M,
highest concentration #4 = 0.0008 M).

Measuring the absorption spectrum and determining λmax

This part of the experiment may be done by all students together, with each pair of students
determining the absorbance at one wavelength. Each pair of students should record all
absorbances at each wavelength and draw the absorption spectrum.

3. Rinse one of the cuvettes with distilled water and fill it with water. Put the cuvette in the
sample compartment. This is the reference solution. Set the wavelength to 400 nm, then set
the Absorbance to zero.

4. Rinse a second cuvette once with distilled water and once with standard solution #1, then
fill it with standard solution #1 (0.0008 M KMnO4). Place the cell in the sample
compartment, measure the Absorbance at 400 nm and record in your notebook.

5. Repeat this procedure (steps 3 and 4 above) for the two cuvettes at wavelengths 420, 440,
460, …600 nm, first setting A = 0 for the cuvette with water, then measuring A for the cuvette
with 0.0008 M KMnO4, recording the absorbance at each wavelength. Record in data table.
6. Prepare a graph of absorbance A vs. wavelength λ and determine λmax (maximum
wavelength). Attach this graph to the lab report.

49
The calibration curve

This part of the experiment must be done by each pair of students separately.

7. Set the wavelength at 525 nm (λmax). Place the cuvette with distilled water in the cell
compartment and again set the Absorbance to zero.

8. Measure and record the Absorbance of each of the four standard solutions, starting with
the most dilute standard. After each measurement, rinse the cuvette with the next standard,
not with distilled water!

9. Draw a plot having X-axis as concentration (mole/L) and Y-axis as Absorbance at λmax
(525 nm).

10. Use Beer’s law to calculate ε for KMnO4, given the cell width (path length l ) to be 1 cm.

Laboratory Data Sheet

1. Stock solution

(1) Mass of KMnO4 = g

(2) Molar mass of KMnO4 = g/mol

(3) Moles KMnO4 = mass/molar mass = mol KMnO4

(4) Molarity of stock solution = mol KMnO4/0.100L= mol/L

2. Preparation of standard solutions

Standard solution 1 (dilute 1.0 mL stock to 10 mL).

KMnO4 molarity = (1.0/10)x(stock solution concentration) = mol/L

Same for concentrations of solutions “2”, “3” and “4” with 500, 200, and 100 μL stock. Remember
that 100 μL is 0.1 mL.

KMnO4 molarity for standard solution 2 = mol/L

KMnO4 molarity for standard solution 3 = mol/L

KMnO4 molarity for standard solution 4 = mol/L

50
3. The absorption spectrum of KMnO4:

S. # Wavelength Absorbance
1 400
2 420
3 440
4 460
5 480
6 500
7 520
8 540
9 560
10 580
11 600

Make a graph of A vs λ at 20 nm wavelength intervals from 400 nm to 600 nm (as measured by the
whole laboratory group). Attach the graph with this lab report.
.
4. The calibration curve at λmax = 525 nm

Standard Concentration Absorbance

1
2
3
4

Make a graph of A (y-axis) vs M (x-axis). Attach the graph to this report.

5. Calculation of the molar extinction coefficient ε at 525 nm.

Use the A-value of solution 1:

ε = A/(C. l), with l = 1 cm ε =

Example: how to make a graph in Excel.

Standard Concentration Absorbance

1 0.00008 0.124
2 0.00016 0.239
3 0.00040 0.614

51
 4 0.00080 1.160

In Excel, enter the concentration values e.g. in cells b1-b4, and enter the absorbance values in
cells c1-c4. Now continue as follows
1. Insert choose scatter then from the types of graphs choose the one with only your data
points. Your graph will appear.
2. Now you want to show the best straight line through the points:
3. On “layout” select “trendline”, choose “linear”. The line will appear but it will not extend
to the y-axis (where x = 0)
4. Now right click on the line. Choose “formal trendline”
5. Under “forecast” click backward and 0.00008. Also click the box “display equation on
chart”

Your chart is now complete, but still in Layout you have to insert the chart title and
horizontal and vertical axis titles (include units, e.g. for the x-axis title write Concentration
(mol/L). The final graph is shown on the next page. Notice the many ways in which Excel
allows you to edit the data points, the trendline, titles, sub- and superscripts, etc.

On the graph you can see that the trend line does not pass exactly through zero. This is as
expected: it may be due to the statistics of the data point, which are not exactly on the straight
line due to random errors in the concentration and/or the absorbance reading, or to the fact
that there is a remaining solution absorbance (relative to the blank) for the standards. For
these reasons, you should not choose the “set intercept” box when formatting the trendline.

52
Finally, note that the trend line slope is the Molar Extinction Coefficient (Molar
Absorptivity)! The equation y = 1442.7x + 0.0149 for this application should be read as
A = ε.b.C + constant, with the constant = intercept.

53
.

FURTHER READINGS/ REFERENCES

1. Barrow,G.M. (1996).Physical Chemistry, [Link]-Hill.

2. Moore,W.J. (1972).Physical Chemistry, [Link].

3. Levine,I.N. (2002). Physical Chemistry, [Link]-Hill

4. Douglas A. Skoog, Donald. West, F. James Holler, Stanley R. Crouch. Fundamentals of

Analytical chemistry, stanfored university, San Jose state university, university of Kentucky and
Michigan state university respectively, 8th Edition, 2004.

5. Christian, G. D. Analytical chemistry, John Wiley and Sons, International Edition: Hoboken,
NJ, 6th Edition., 2004.

6. Harris, D. C. Quantitative Chemical Analysis, 4thed., WH Freeman: New York, 1995.

7. NigussieReta (Ph.D). Quantitative Chemical Analysis, Experiments For University Students,

Addis Ababa University, Revised and Second Edition, 2000.