Experiment (2)

Spectrophotometric Determination of Tin(IV) with Catechol-Violet and

Cetyltrimethyalmmonium bromide

Theory:

Tin(IV) forms a colored 1:2 complex with Catechol-violet [C19H13O7SNa2] which can be
used in colorimetric analysis of tin.
Unfortunately, the spectra of both, the reagent (catechol-violet) and the complex are
similar and overlapping could occur, due to the unreacted reagent.

Fortunately, it was found that in the presence of Cetyltrimethyalmmonium bromide

[C16H33(CH3)3NBr], the maximum absorption of the complex (but not the reagent) is
shifted to a longer wavelength (red shift) and has a much larger intensity.

The absorption spectra of the complex will be plotted in the presence and in the
absence of CTMABr, in order to study the effect of this reagent on the measurements.
When the proper maximum wavelength is determined, a calibration curve obeying
Beer's law is constructed and used to determine the concentration of tin(IV) in an
unknown sample.

Chemicals:
1) Tin(IV) solution; 1x10-5 M
2) Catechol-violet solution; 0.040% in water [wt/wt].
3) Cetyltrimethyalmmonium bromide solution, 0.1% in water [wt/wt].
4) Acid solution of PH 2.2.
5) Conc.H2SO4 and cont.NH3 to adjust to PH 2.2 condition.
6) Unknown sample solution.
Apparatus:
1) Spectrophotometer
2) Cuvettes (2)
3) Burettes (2), 50 mL
4) Volumetric flasks (10), 100 mL
5) Beakers (10), 50 mL

Procedure:
Prepare different solutions' as shown in the following table:

SOLUTION CATECHOL H2O TIN CTMABR ADJUST FINAL

NUMBER VOLLUME VOLUME VOLUME VOLUME
VOLUME WITH ACID
SOLUTION
1 10 40 -- -- THE PH OF 100
2 10 40 10 -- THE 100
3 10 40 0.0 4 SOLUTION 100
4 10 40 2.5 4 TO 2.2 100
5 10 40 5.0 4 WITH 100
6 10 40 7.5 4 DIL.H2SO4 100
AND
7 10 40 10.0 4 100
DIL.NH3
8 10 40 12.5 4 100
SOLUTION
UKNOWN 10 40 5 4 (PH 2.2) 100
1)Scan* solution (7) from 400 nm to 700 nm in 20nm steps.

2) Determine the maximum absorbance (in the laboratory) from the results of solution
(7), then record the absorbance of the solutions (3 to 8) at the wavelength of
maximum absorbance you have determined.

3) Scan solution (1) form 400 nm to 700 nm to 20 nm steps.

4) Scan solution (1) form 400 nm to 700 nm to 20 nm steps.

*scanning means recording the absorbance of a solution starting at a certain

wavelength and increasing or decreasing the wavelength with a certain speed and the
absorbance after each variation.

Treatment of results:

1) Make the needed absorbance corrections using solution (3).

2) Plot A (absorbance) vs. (wavelength) for solution (1) and (2) and comment on the
findings.
3) Plot A (absorbance) vs. (wavelength) for solution (7) to determine the maximum
absorbance wavelength.
4) Using the standard calibration curve constructed form the results of the solutions
(3 to 8); determine the unknown tin concentration.
5) Discuss your results and the role of using CTMABr.
6) Use the least square method for drawing all the calibration curves to get the best
line. 

Questions:

1) Write down the structure of the colored complex formed in this experiment.
2) Account for the red shift that takes place by the addition of CTMABr.
3) What is limiting reagent in solutions (3 to 8) ?
4) If Beer's law is applicable, calculate the value of molar absorptivity for every
solution in 7.

Experiment (6.I)
Polarimetry / Optical Activity
 Theory
Introduction:
According to wave theory of light, an ordinary ray of light is considered to be vibrating
in all planes at right angles to the direction of its propagation. If this ordinary ray of
light is passed through a Nicol prism (a special prism obtained by combining tow
prisms of Iceland spar suitably cut), the emergent ray has it's vibrations only in plane.
This light having wave motion in only one plane it's known as plane polarized light
(PPL), (Fig.1).

The plane along which vibrations are taking place is known as plane of polarization.
The Nicol prism used for this purpose is called the polarizer. It is interesting to note as
to what happens when a plane polarized light is passed through and reaches the eye
as shown in (fig.2).
If however, the axis of second Nicol prism is perpendicular to the axis of the first the
polarized light is totally cut off due to total internal reflection and no light reaches the
eye. In the remaining position of the second Nicol prism, the polarized light is partially
transmitted. This second Nicol prism used is known as analyzer.

Optical activity:

When certain organic liquids, solution (like sugar) or quartz crystals are placed in the
path of plane polarized light, the plane of polarization is rotated. The property by virtue
of which the plane of polarization of light is rotated is called optically activity and the
substances possessing this property are said to be optically active. Substances which
rotate the right (clockwise) are called dextro rotatory (+) while substances which
rotate the light towards left (anticlockwise) are called levo rotatory (-). A mixture of
these two varieties in equal proportions will be optically inactive and is called racemic
mixture.
It has been found that the magnitudes of the rotation depend upon the following
factors:
1) Nature of substance.
2) Length of the cell.
3) Concentration of the solution.
4) Nature of the solvent.
5) Temperature of the solution.
6) Wavelength of light used.

The rotation power of a given solution is generally expressed as specific rotation, []Dt.
It is the number of degrees of rotation of the plane polarized light produced by one
decimeter in length filled with a solution having one gram of the substance per mL.
The measurement is carried out at a temperature (t) and using the D-line of sodium
light.
The specific rotation can be calculated by the following relation:

[α]Dt =

It should be kept in mind that for very dilute solutions in any given non-active solvent,
the specific is constant.

(Note: read about optical activity and optical isomerism and about polarimeter) 1,2,3
Experimental procedure:-

1) Make up a series of 4 standard solution of sucrose in water by weighing out

approximately 1,2,3 and 4 g of sucrose (to the nearest 0.01g) and making up to 100
mL with distilled water in a volumetric flask.

2) Open one end of the polarimeter tube by unscrewing the cap. This operation must
be done carefully over a piece of tissue paper placed on the bench. Be careful not to
allow the glass disc to fall out, otherwise it may break.

3) clean the glass disc with a piece of tissue paper. Wash out the tube with distilled
water.

4) Holding the tube vertically, fill it with distilled water so that the level of liquid is
slightly above the rim of the tube. Slide the glass disc and screw the cap into place.
You must tighten the cap sufficiently to prevent leakage, but do not overtighten
otherwise the glass disk may break.

5) After making sure that there is no liquid on the outside of the glass discs and no air
bubble inside the tube, place the tube in the polarimeter and observe the optical
rotation. If the reading is not zero then the zero error of the instrument should be
noted.

6) Repeat steps 2 to 5 by using the first sucrose solution and observe the rotation.
Correct for the zero error if necessary. Repeat using the other solutions.

7) After finishing your measurements, note the room temperature. Make sure that the
tube is left in very clean state otherwise the caps may stick and damage will be
caused.

Treatment of results:

Make a table containing the different concentrations of sucrose and the related
observed angels of rotations.

Calibration graph is obtained from plotting θ against C. The slope of this graph 

represents the specific rotation of sucrose. 

Concentration of the unknown could be obtained form the calibration graph.

The least square method should be applied here and statistical data should be 
obtained.
Question:

1) Draw the structure of sucrose and discuss its optical activity ?

2) Write brief notes on the following:

 (a) Polarized light.
(b) Racemic mixtures.
(c) Plane of polarization.

3) A substance has a specific rotation at 20 oC in water of +66.5, what would you

expect the value of [α]Dt to be (larger, smaller or equal) at the following conditions:

A) The substance under goes hydrolysis in the presence of acid getting two
fragments.

B) Measurement was done at:

a- 30oC, same concentration in water.

b- 20oC, twice concentration in water.
c- 20oC, same concentration in alcohol.

4) What are the factors affecting [α]Dt ?

5) An isomer (+) show [α]Dt = +5.2o, what would you expect its (-) isomer to has as
[α]Dt ?

Experiment (6.II)

Refractometry

Theory:

When a ray of light passes from air into a denser medium such as glass or a liquid, it
bends towards the normal as in fig.1.
 Sin i / sin r = constant = n,

where, i ≡ angle of incidence, angle of reflection.

The constant n is called "refractive index" of the medium. Refractive index of a

substance is ordinarily determined by measuring the direction (refraction) of
collimated radiation as it passes from one medium to another

n /n = v /v = sinθ
2 1 1 2 1 / sinθ2

Where v1 is the velocity of propagation in the less medium M 1 and v2 is the velocity in
medium M ; n and n are the corresponding refractive indexes and θ and θ are the
2 1 2 1 2

angles of incidence and refraction, respectively. When M is a vacuum, n 1 1 is unity because v1 becomes equal to

C (light velocity) as follows

n
2 = nvac. = C/v = sin θ / sin θ2
2 1

where navc. is the absolute refractive index of M2. Thus, nvac. Can be obtained by measuring the two
angles θ and θ2. 1

It is much more convenient to measure the refractive index with respect to some 
medium other than vacuum, air commonly serves as a standard medium for this 
purpose.
 
Refractive index is one of the common physical constants that can be used to 
describe a chemical species.

Most of complication of n for liquids and solids in literature refer to air laboratory
temperature and pressures. Fortunately, the change in the refractive index of air with
respect to temperature and pressure is small enough so that a correction from
ambient laboratory condition to standard conditions is needed for the most precise
work. A Refractive index nD measured with respect to air using the radiation of the D
line of sodium can be converted to nvac. With the equation:
nvac = 1.00027 nD

This conversion is seldom required. It is usually necessary to measure a refractive

index to an accuracy of at least 2 x 10-4.

Refractometery is based on the measurement of the so-called critical angle. The

critical angle is defined as the angle of refraction in a medium when the angle of
incident radiation approaches 90 degrees, θ2 becomes the critical angle (θc).
Thus
n2/n1 = sin 90/ sin θc = 1/ sin θc

It is important to realize the critical angle depends on wavelength. Thus, if

polychromatic radiation is employed, no single sharp boundary will be obtained, which
makes precise establishment of the critical angle impossible.

This difficulty is often overcome in refractometers by the use of monochromatic

radiation, one is sodium D-lines.

The refractive index of a liquid can be easily determined by Abbe's refractometer

(Fig.2.). It consists of two prisms A and B held in contact

with each other with a clamp. The surface of the upper prism B in contact with A is
polished white the surface of A in contact with B ‫ﺗﻼﻧﺘﻼﻧﺘﻼ‬
 .(layer sample the to rays of number infinite an introduce to )

 The .telescope the below fixed axis horizontal a about rotated be can prisms The
 to scale the over moves which ,arm an with attached is prism the leaving case metal
 water a in enclosed further is system prism The .directly index refractive the read
.temperature the control to jacket

Refractometric measurements are used for qualitative and quantitative analysis

as well as for structural investigations.

It has been found that the molar refraction increases in regular increments
with the number of carbon atoms within a homologous series. This finding
has led to the notation that the molar reflection of the compound can be
considered to be the sum of atomic increments and that within certain limits
the contribution of each atom is the same in every molecule.

The molar refraction of a few important atoms are given in the following
table:

Group r
M Atoms M
r Bonds M
r
or linkages
4.60 C
2.55 H
7.25 Cl
4.71 Br
I
5.65 O in
–OH
10.3 O in 1.6
R-O-R 4
4.65 O in 2.21
>C=O
Example:

The kekule's formula for benzene is

So, molar refraction due to 6C atoms is 6 x 2.42 = 14.52,

Molar refraction due to 6H atoms is 6 x 1.10 = 6.60,

Molar refraction due to 3 double bonds 3 x 1.73 = 5.19,

Molar refraction due to one six membered ring is 1 x 0.15 = 0.15,

_________________________________________________

Total M = 26.46
r

The experimental value , 25.95 is fairly close to the above calculated value.

Chemicals:
1) Distilled water,
2) Benzene,
3) Acetone,
4) Methanol,
5) Chloroform,
6) n-Hexane.
Apparatus:
Abble's refractometer (Fig .2).

Procedure:
1) Using a rounded end glassrod, introduce on the lower prism a large
drop of distilled water. Close the prisms carefully, clamp them and
adjust the refractometer so that two zones appears, one is shadowed
and adjust the bright. Move the prisms by rotating the scale until the
light-dark boarder is in the center of the refractive index.
2) Do the same for the other five solvents.
 3) Measure the refractive index of the unknown in the same way.
4) Before you leave the laboratory record the density and the molecular
weight for each substance and the laboratory temperature.
Treatment of result:
1) Construct a table containing the name, molecular weight density,
refractive index, experimental molar refraction, and calculated molar
refraction for each known substance. (Show sample calculation).
Specific refraction r =

Molar refraction R = r.M , where M=M.wt

d= density

2) Calculate the experimental molar refraction of the unknown and

according to the formula, suggest at least three structures and then
calculate the molar refraction for each structure.

3) Compare the values of experimental and calculated molar refraction for

each suggested structures.

4) Comment on the data obtained, and show which of the suggested

structures the most convenient one is.

Questions:

1) Discuss the variables which affect the measurement of refractive

index.
2) Suggest reasons why quantitative analysis by refractometry is
restricted to studies of binary mixture.
3) What is the wavelength of sodium D-line.
 Experiment 12
Ion exchange Chromatography

Theory:

Ion exchange chromatography is based on the exchange of ions

between a polar liquid mobile phase and stationary solid phase with
ion-exchange groups. The groups are generally chemically fixed on an
insoluble cross-linked polymer called resin.

Cations can be separated on cation exchanger resins which contain

negatively charge functional groups such as –SO-3or
-COO-.

rged functional groups such

Anions can be separated on anion exchanger resins which contain positively cha

as –CH N (CH ) , a quaternary ammonium ion.

2
+
3 3

Separation is based on ions-partitioning into the ion-exchanger phase

to different degrees. The selectivity of a resin for an ion is determined
primarily by the change of the ion and its hydrated radius. Resin
affinity increases with increasing the charge density.

Zn and Mg ions can be separated using either cation- or

2+ 2+

anion-exchenger.

In this experiment, and although both are cations, Zn and Mg are

2+ 2+

going to be separated by anion-exchanger. This is possible because

Zn with an excess of Cl- forms an anionis complex ZnCl , while Mg
2+
4
-2 2+

does not forms such an anionic complex.

Chemicals :

1) Buffer solution (PH=10)

2) 2 M HCl solution.
3) 0.25M NaOH solution.
4) 0.01M EDTA.
5) Erichorm Black T (EBT), as an indicator.
6) Unknown mixture of and .
7) Distilled water.
8) Anion exchanger resin.

Apparatus:

1) Glass column.
2) PH meter.
3) Burette.
4) Erlenmeyer flask.
5) Pipette.

Procedure:

1. Regenerate the anion-exchanger column with 50ml of 2M HCL.

2. Pipette 5.0ml of the unknown solution onto the top of the column.
3. Wash the column with 70ml of 2M HCL and collect the eluates in a
100ml Erlenmeyer flask labeled "Mg".
4. Wash the column with 70ml of 0.25M HNO3 and collect the
eluates in a 100ml Erlenmeyer flask labeled "Zn".
5. Adjust the PH of each solution to be between 9 and 10, then add
10ml of the buffer solution of the PH 10.
6. Add 5 drops of EBT-indicator and titrate vs. 0,01M EDTA.
7. Regenerate the column with 2M HCL.
Treatment of results :

1. Tabulate your results.

2. Determine the concentration of both Zn2 and Mg in the unknown
+ 2+

sample.
Question:
1. EDTA is a good complexion agent for metals. Which conditions are
necessary to perform a successful titration reaction for the
determination of a metal concentration? Give reactions!

2. What is the role of using HNO3 solution in this experiment ? Give reaction
!

3. What is the color of EBT-indicator at different pH values , especially

acidic and basic ?

4. In the mixture that contains Ca2+ , Zn2+ and H3O+ ions, describe
briefly how can you determine the concentration of each ion ?

Experiment 13

Adsorption Chromatography

Theory:

Adsorption Chromatography utilizes a solid stationary phase of a

polar nature such as silica or alumina.

The mobile phase and the solute (components of the sample ) are in
competition for the active adsorption sites on the stationary phase
particles.

Thus, strongly adsorbed components are retained longer than weakly

adsorbed component.

Because more polar compounds adsorb on a polar surface to a

greater degree than do less polar compound, retention on the column
and thus retention time is related to solute polarity.

Dichromate and permanganate ions can be separated using Alumina

due to their different polarities.
Chemicals :

1) 0.5N HNO 3

2) 2N H SO
2 4

3) 0.02N Ferrous Ammonium sulfate.

4) Alumina.
5) Unknown aqueous solution mixture of KMnO and K2Cr O . 4 2 7

6) 0.02N KMnO (standard solution).

4

Apparatus:
1) 30cm column (i.d.=1 cm )
2) Erlenmeyer flask, 100ml (2).
3) Beaker, 100ml (2).
4) Volumetric flask, 100ml(2).
5) Pipet, 10ml.

Procedure:

A. preparation of the column

1. Prepare a slurry of alumina by adding 10g of alumina and
30ml of 0.5N HNO . 3

2. Transfer the slurry to the column using 0.5N HNO . 3

B. Separation process:
1. Add the whole unknown sample solution on the top of the
column and let the unknown to penetrate the alumina
surface..
2. Wash the column with 0.5 N HNO until the purple color of
3

permanganate fraction approaches the column lower end.

3. Elute the permanganate fraction in a 100ml Erlenmeyer
flask by washing with 0.5 N HNO (total volume = 80ml).
3

4. To another Erlenmeyer flask, elute the dichromate fraction

using 2 N H SO in the same manner.
2 4

5. Dilute each eluted fraction to 100 ml using volumetric

flasks.

C. Determination of KMnO and K Cr O concentrations:

4 2 2 7

Back-titrate each fraction using the following procedure:

a. To each of three Erlenmeyer flasks pipet 10.0ml of each

fraction.
b. Using a buret (or a pipet), add an excess of 20.0ml of
 0.02N Fe(NH )SO to each flask.
4 4

c. Titrate the excess of Fe2+ in each flask vs. 0.02N KMnO . 4

Treatment of results:
Determine the concentration of the unknown for KMnO 4

and K Cr O as grams per 100ml. Show all calculations.

2 2 7

Questions:
1) What is the purpose of using 2 N H SO ?
2 4

2) According to the structure of alumina, explain the

behavior of both KMnO and K Cr O in this
4 2 2 7

experiment.
3) Write down the equation for the reactions that take
place in the back-titration process.