Engineering Chemistry Laboratory

Fall Semester 2024-2025

Course Objective
To apply theoretical knowledge gained in the theory course and get hands-on experience of the
topics
Course Outcome (CO)
Understand the importance and hands-on experience on analysis of metal ions, synthesis of
organic molecule and nanomaterials and invoking the concepts of thermodynamics and kinetics
Table of Contents (Experiments)

Sl. No. Indicative Experiments Page No.

1 Thermodynamic functions from EMF measurements : Zn – Cu system 3
2 Determination of reaction rate, order and molecularity of ethylacetate 9
hydrolysis
3 Colorimetric estimation of Ni2+ using conventional and smart phone 13
digital-imaging method
4 Laboratory scale preparation of important drug intermediate – Synthesis 17
of 2,3-Diphenylquinoxaline
5 Estimation of sulfate ion in drinking water by conductivity method 21
6 Analysis of iron in an alloy sample by potentiometry 25
7 Preparation of nanosilica by sol-gel method and its characterization 31
8 Determination of hardness of water sample by complexometric titration 35
before and after ion-exchange process
Demo experiments
9 Size dependent colour variation of Cu2O nanoparticles by 40
spectrophotometer
10 Computational Optimization of molecular geometry using Avogadro 44
software

Important Instructions:

 Students are advised to take printout of this manual along with graph sheets and use
them as the observation note.
 After entering the experimental data/plotting of graphs/calculation of final result in
the observation note, students should take the picture/scan of the same and upload
it in the course page within a week of each completed lab slot.
 Total = 60 M (based on 8 regular experiments x 7 M+ Quiz on 8 regular experiments
4M) = 60 M) QUIZ: 20 Questions x 0.5M=10M (minimum 2Q from each experiments)
 FAT: Marks distribution = 40 M (Experiment) + 10 M (Short Procedure) = 50 M
(Weightage = 40 M).

Page | 1
 Index Table

Sl. No. Experiments Obtained Marks Signature of Faculty

1 Thermodynamic functions from EMF
measurements : Zn – Cu system
2 Determination of reaction rate, order
and molecularity of ethylacetate
hydrolysis
3 Colorimetric estimation of Ni2+ using
conventional and smart phone digital-
imaging methods
4 Laboratory scale preparation of
important drug intermediate –
Synthesis of 2,3-Diphenylquinoxaline
5 Estimation of sulfate ion in drinking
water by conductivity method
6 Analysis of iron in an alloy sample by
potentiometry
7 Preparation of nanosilica by sol- gel
method and its characterization
8 Determination of hardness of water
sample by complexometric titration
before and after ion-exchange process

Page | 2
Expt. No.: Date:

Experiment Thermodynamic functions from EMF measurements: Zinc-Copper system

To construct and to measure electromotive force (EMF) of a Daniell cell at varied

Aim concentrations of metal ions and temperatures
To determine the thermodynamic functions (∆G, ∆H and ∆S)
Problem
Construction of a Daniell Cell, measurement of EMF and determination of ∆G, ∆H & ∆S
definition

The measurement of EMF will be carried out by a Potentiometer and followed by the
Solution
calculation of different thermodynamic functions through the EMF values

Students will be able to understand the construction of a Daniell cell (Zn-Cu system)
Student
learning  To observe the influence of metal-ion concentration and temperature on the EMF
outcomes on Zn-Cu system
 To calculate the thermodynamic functions (∆G, ∆H and ∆S)

Principle:
The electromotive force (EMF) of an electrochemical cell is measured by means of a
potentiometer. An electrochemical cell (Ecell) is considered as a combination of two individual
single cell-electrodes. The potential difference between the two single electrode potentials is
a measure of EMF of the cell (Ecell; Cathode-Anode/V). In order to measure the potential
difference between electrodes in contact with electrolyte containing the same cation, it is
necessary to have another electrode in contact with electrolyte of same cation, both the half-
cells connected through a salt bridge. Saturated calomel electrode (SCE; ECalomel, Eo = 0.244 V)
whose potential is known, is used as a reference electrode (Anode; +ve terminal, black-
colored wire) and it is coupled with the metal electrode (Cathode, -ve terminal; red-colored
wire) for which the potential is to be determined. The substance that loses electrons is said
to be oxidized, while the one gaining electrons is reduced. Thus if a piece of zinc metal were
immersed into a solution containing copper (II) ions, zinc would be oxidized by copper (II) ions.
Zinc loses electrons and is oxidized, and the copper (II) ions gain electrons and are reduced.
We can conveniently express these processes together as a Daniel cell by the following
reactions (Figure 1):
In the Daniell cell, copper and zinc electrodes are immersed in the equimolar solution of
copper(II) sulfate and zinc(II) sulfate respectively. At the anode, zinc is oxidized as per the
following half-reaction:
Zn(s) → Zn2+(aq) + 2e− (Eo = -0.760 V)
At the cathode, copper is reduced as per the following reaction:
Cu2+(aq) + 2e− → Cu(s) (Eo = +0.340 V)

Page | 3
The overall reaction:
Zn(s) + Cu2+(aq) → Zn2+(aq) + Cu(s) Ecell = 0.340 – (-0.760) = 1.10 V

Figure 1: Functioning of a Daniell cell (Zn-Cu) using a salt bridge

Importance of Thermodynamic functions:

∆G: Gibbs free energy determines whether a given chemical change is thermodynamically
possible or not. The change in Gibbs free energy (∆G) is the symbol for spontaneity, and there
are two factors which can affect it: enthalpy and entropy. Enthalpy - the heat content of a
system at constant pressure. A negative ∆G means that the reactants, or initial state, have
more free energy than the products, or final state.
∆H: Measuring the change in enthalpy allows us to determine whether a reaction was
endothermic (absorbed heat, positive change in enthalpy) or exothermic (released heat, a
negative change in enthalpy.) It is used to calculate the heat of reaction of a chemical process.
∆S: Entropy is the measure of a system's thermal energy per unit temperature that is
unavailable for doing useful work. Entropy has been regarded as a measure of disorder or
randomness of a system. Thus when a system goes from a more orderly to less orderly state,
there is an increase in its randomness and hence entropy of the system increases.
Part-A: Determination of single electrode potentials of M/Mn+ system (EoM/Mn+) at three
different concentrations
Hg/Hg2Cl2 (s), saturated KCl (Anode)║(N/10) electrolyte of the Metal/Metal (Cathode)
From the EMF of the cell involving saturated calomel electrode and metal electrode dipped
in its solution of 0.1 M electrolyte, electrode potential of the metal electrode is readily
calculated using the standard potential of calomel electrode as Ecell (measured from
potentiometer) = EM/M+ (Cathode, V) – Ecalomel (0.244V, Anode) or EM/M+ = Ecell + Ecalomel. Ecell is
the total EMF, i.e., net voltage of the cell measured using a potentiometer. Then, the
electrode potential of the metal electrode (EoM/M+/ V) can be calculated after incorporating
the observed EM/M+ and aMn+ parameters in Nernst equation as given below:
𝑅𝑇
EM/M+ = EoM/M+ + 𝑛𝐹 aMn+
𝑅𝑇
or, EoM/M+ = EM/M+ – 𝑛𝐹 ln aMn+

Page | 4
 0.059
EoM/M+ = EM/M+ – log aMn+
𝑛

Requirements:
Reagents and solutions: Copper sulphate stock solution (0.1 M), Zinc sulphate stock solution
(0.1 M), KCl salt.
Apparatus: Digital potentiometer, copper electrode, zinc electrode, calomel electrode, 100
mL beaker, burette and standard flask
Procedure:

Calibrate the digital potentiometer with the help of an inbuilt battery support to display 1.018
V. The metal electrode is sensitized by dipping in a small quantity of 1:1 nitric acid containing
a small quantity of sodium nitrite until effervescence occurs. Then the electrode is washed
well with distilled water. 50 mL of the given concentration of the electrolyte solution is taken
in a beaker and its corresponding metal electrode is introduced. This is connected with the
saturated calomel electrode (half-cell) by means of a salt bridge. The metal electrode is
connected to the positive terminal of and the calomel electrode is connected to the negative
terminal of the potentiometer. The EMF of the cell (Ecell) is measured and noted in Table 1.
The standard electrode potential [E°M/M2+] is computed using Nernst equation (Eq. 1).
Table 1: EMF measured for various concentrations of M/Mn+ system

Electrode/ Concentration of Ecell (V) EM/M+ EoM/M+/V (from

Nernst Eq.1)
Electrolyte Electrolyte (N) (EMF of the cell) = Ecell + Ecalomel
Zn/Zn2+ 0.05 M
0.1 M
Cu/Cu2+ 0.05 M
0.1 M

Table 2: Individual activity coefficients of Cu2+ and Zn2+ in water at room temperature

Metal ion (Cu2+/Zn2+)

0.001 0.002 0.005 0.01 0.02 0.05 0.1 0.2
system (in M)

Activity coefficient (γc) 0.905 0.870 0.809 0.749 0.675 0.570 0.485 0.405

where, E° is the standard electrode potential of the metal electrode; a Mn+ is the activity of
metal ions in the solution (aMn+ = γc[C]); γc is the activity coefficient (Table 2) and C is the
concentration of the electrolyte solution. Solution Temperature (T) = 30 °C

Page | 5
The potential of saturated calomel electrode (SCE) = 0.244 ± 0.0007 (30 °C)
𝟎.𝟎𝟓𝟗
E°M/M+ = EM/M+ – Iog [γc x C] ................Eq.1
𝒏

The EMFs can be calculated by the Nernst equation (Eq.1).

Part-B: Construction of Daniell cell and measurement of its voltage with three different
concentrations of Copper and Zinc Salt Solutions:
The construction of the Daniell cell using the following concentrations of Copper and Zinc salt
solutions will be performed and the voltage of the cells will be entered into the Table 3.
Calculation of Ecell by Nernst Equation
For example, if [Zn2+] and [Cu2+] are given as 0.05 M and 0.01 M, respectively, then
[Zn2+] R = 8.314 J· K-1·mol-1;
RT
ECell = ECello_ ln T = 303 K;
nF [Cu2+] n = 2;
F = 96500 (constant)

8.314 x 303 ln [0.05]

= 1.1 _
2 x 96500 [0.01]

[0.05]
= 1.1 _ 0.01305 ln
[0.01]

= 1.1 _ (0.01305 x 1.6094)

= 1.1 _ 0.021
ECell = 1.079 V at T = 303 K

In a similar way, Ecell values for other concentrations are to be calculated at specific
temperatures.
Calculation of ∆G1 (at T1 = 30 oC) and ∆G1a (at T1a = 50 oC) based on the ECell measurements

In a similar way, ∆G2, ∆G2a, ∆G3, and ∆G3a are to be calculated from EMFs for other metal ion
concentrations.

Page | 6
Table 3: EMF of Daniell Cell observed (by calculation and experiment) from two different
concentrations of zinc and copper solutions

Free-energy
Ecell (calculated Ecell
[Cu2+] [Zn2+] % Error change (∆G) or
by Nernst Eqn) (experimental)
Wmax (KJ/ mol)
At Room Temperature, T1 = 30 ºC/ 303 K

0.1 M 0.05 M

0.05 M 0.1 M

0.1 M 0.1 M

At T1a = 50 ºC/ 323 K

This value will
0.1 M 0.05 M be provided by
the lab faculty.
This value will
0.05 M 0.1 M be provided by
the lab faculty
This value will
0.1 M 0.1 M be provided by
the lab faculty

Part-C: Calculation of ∆H and ∆S at 40 ºC / 313 K based on the T1 and T1a parameters

∆G1 at 30 ºC = -191 KJ/ mol and ∆G1a at 50 ºC = -197 KJ/ mol, Then

∆G2 at 30 ºC = -195 KJ/ mol and ∆G2a at 50 ºC = -199 KJ/ mol, Then

∆G3 at 30 ºC = -206 KJ/ mol and ∆G3a at 50 ºC = -208 KJ/ mol, Then

Page | 7
According to the Gibbs-Helmholtz equation, the ∆H can be calculated for Daniell Cell having
[Zn2+]/[Cu2+] = [0.05 M]/ [0.01 M],

,
then

In a similar way, the ∆H and ∆S for other concentrations of [Zn2+] and [Cu2+] as given in Table-
4 are to be calculated.
Table-4: Final Results: At 313 K (40 ºC)

∆S, Marks
[Cu2+] [Zn2+] ∆G, KJ/ mol ∆H, KJ/ mol
KJ⋅K−1⋅mol−1 Awarded

0.1 M 0.05 M

0.05 M 0.1 M

0.1 M 0.1 M

Page | 8
Determination of reaction rate, order and molecularity of hydrolysis of ethyl
acetate
(KINETICS OF ACID CATALYSED HYDROLYSIS OF ETHYL ACETATE)

Introduction:
Chemical reactions and reaction rate

Chemical kinetics is the part of physical chemistry that studies reaction rates. The reaction
rate for a reactant or product in a particular reaction is intuitively defined as how fast a
reaction takes place. Through the study of chemical kinetics, one can reasonably get an idea
as to how to alter the reaction conditions and improve the reaction rate, which is necessary
to increase the production of chemical products in the industry perspective. It will also help
us to learn how to suppress or slow down unwanted side reactions.

For a generic reaction: A + B C

the simple rate equation is of the form:
The concentration is usually in mol cm-3 and k is the reaction rate coefficient or rate constant.
Although it is not really a constant, because it includes everything that affects reaction rate
outside concentration: mainly temperature, ionic strength, surface area of the adsorbent or
light irradiation (in the case of photochemical reactions).
The exponents a & b are called reaction orders and depend on the reaction mechanism. The
stoichiometric coefficients and reaction orders are very often equal, but only in one step
reactions.

Acid-catalyzed Hydrolysis of Methyl Esters

The hydrolysis of ester is catalyzed by either an acid or base. This can be achieved in a number
of ways. The most common method is to use a Lewis acid or Bronsted acid to form a positively
charged intermediate that is far more reactive and even mild nucleophiles such as water will
react.

Page | 9
 Expt. No.: Date:

Determination of Reaction Rate, Order and Molecularity of

Experiment
Hydrolysis of Ethyl Acetate
Hydrolysis of ethyl acetate in acid medium to produce acetic acid and
Problem definition
ethyl alcohol
Following chemical kinetics of the reaction through acid-base
Methodology
titrimetric analysis
The formation of acetic acid in the hydrolysis of ethyl acetate will be
Solution
followed by a titration with NaOH at different time intervals.
Student learning Students will be able to calculate the rate constant of ester hydrolysis
outcomes through (pseudo) first order kinetics.

Aim:
To determine the rate constant of the hydrolysis of ethyl acetate using an acid as a catalyst
and to understand the order and molecularity of the reaction
Principle:
The hydrolysis of an ester occurs according to the equation:

The following rate equation is applicable to the above reaction:

Rate  [Ester] [H2O]
Since [H2O] remains constant,

Rate =  dc  k '1 [ Ester ]

dt
where, ‘c’ represents concentration of the ester at any time t; k’1 is the specific velocity
constant. As the reaction progresses, the accumulation of acetic acid increases. Drawing a
known volume of the reaction mixture at known regular intervals of time and titrating it
against standard sodium hydroxide solution will indicate the increase in acetic acid presence.
The acid hydrolysis of ester is a first-order bimolecular reaction, and the reaction follows
pseudo first order kinetics. This is because the amount of water is in large excess so that its
concentration does not change significantly to alter the reaction rate. The reaction goes
practically to completion (the equilibrium shifts to the right) and the rate is first order with
respect to the ester.
Requirements:
Reagents and solutions: Ethyl acetate, 0.5N HCl, 0.2N NaOH, Phenolphthalein indicator, Ice-
cubes

Page | 10
Apparatus: Burette 50mL, Pipettes-5mL, 10mL, Conical flasks, Wash bottle Reaction bottle
250mL.
Procedure:
Exactly 100 mL of 0.5N-hydrochloric acid solution are taken in a 250 mL clean reaction bottle
and exactly 5 mL of the ester is added to it. Zero time is noted when half the volume of ester
solution in the pipette is transferred into the reaction bottle. After thorough mixing,
immediately 10 mL of the solution is pipetted out into a clean conical flask containing ice-
cubes. It is then titrated against 0.2 N sodium hydroxide solution from the burette using
phenolphthalein indicator. The end-point is the first appearance of a pale permanent pink
colour. The same volumes of the reaction mixture are withdrawn at regular intervals, say 10
minutes and is titrated against sodium hydroxide solution. The reaction is allowed to go to
completion by keeping the reaction mixture over a hot water bath for about 90 minutes. The
final reading is then noted.
Calculations:
Let V0 be the volume of alkali used at zero time and Vt. be the volume of alkali used after the
time ‘t’ seconds. Let V be the reading when the reaction is completed. Concentrations of the
ester at various time intervals are expressed in terms of volume of NaOH solution.
a= initial concentration of ester = (V - Vo)
(a-x) = concentration of ester at any time ‘t’ = (V - VO) – (Vt – Vo)
= (V - Vo – Vt + Vo)
= (V - Vt)
The specific rate constant of the reactions is given by

2.303  a 
k1 ' = log  
t a x

 V  Vo 
k1 ' =
2.303
log  
t  
V  Vt 

k1 ' = Slope x 2.303

The rate constant values are calculated at different time intervals which should nearly be the
same. A graph is drawn between log(V - Vt) and time ‘t’. From the slope of the plot, the rate
constant is calculated, and it is compared with the experimental value.

Page | 11
Table-1:
Volume of
S.No. Time in mL
NaOH (mL)
(min) min-1
1 0
2 10
3 20
4 30
5 40
6 ∞

Results:
The Rate Constant for the hydrolysis of an ester from
1. Calculated value =
2. Graphical value =
3. Molecularity of the reaction =
4. Order of the reaction =

Page | 12
Colorimetric estimationn of Ni2+ metal ions using conventional and smart
phone digital-imaging methods

1. Importance of the experiment:

Nickel is a transition element and commonly exists in +2 oxidation state, though +1, +3 and
+4 states are also observed in nickel complexes. Nickel plays an important role in biological
systems as a constituent of several enzymes. Nickel is also present in soils and plants, and its
concentration varies widely from trace quantities to being a major constituent. Therefore,
determination of nickel at different concentration levels in variety of samples becomes very
important.

2. Nickel Toxicity: Compared with other transition metals, Nickel is a moderately toxic
element. However, it is known that inhalation of nickel and its compounds can lead to serious
problems, including cancer in the respiratory system. Moreover, Nickel can cause a skin
disorder known as nickel-eczema (10.1016/j.kijoms.2016.08.003).

3. Nickel in Industries: A thin layer of nickel onto a metal object can be decorative, provide
corrosion resistance, wear resistance, or used to build up worn or undersized parts for salvage
purposes. Nickel alloys are used extensively because of their corrosion resistance, high
temperature strength and special magnetic and thermal expansion properties.
The major alloy types that are used are:
 Iron-Nickel-Chromium alloys
 Stainless Steels
 Copper-Nickel alloys and Nickel-Copper alloys
 Nickel-Chromium Alloys
 Nickel-Chromium-Iron alloys
 Low Expansion Alloys
 Magnetic Alloys (http://www.nickel-alloys.net/nickelalloys.html)

Page | 13
 Expt. No.: Date:
Colorimetric estimation of Ni2+ metal ions using conventional and
Experiment
smart phone digital-imaging methods
Corrosion protection in steel depends on the amount of Ni (acts as
Problem definition passivating metal) in its composition. Hence, it is important to analyze the
amount of Ni in steel for its use in industry.
Ni-DMG forms a stable colored complex. With increasing concentration of
Ni in solution, its color intensity also increases. In turn, the color intensity
Methodology
is a function of color coordinates (Red, Blue and Green, RGB) in the image
taken using mobile phone camera.
Estimation of Ni concentration in the unknown sample from the
Solution
calibration graph plotted based on different known Ni concentrations.
Student learning Students will learn to perform colorimetric method, perform RGB
outcomes response analysis and analyze Ni composition in different grades of steel

(i). Principle:
(a) Colorimetric method:
Photo-sensitive measurements are expressed in terms of absorbance, (A) as given in Eq. (1).
Further, the linear relationship between absorbance (A) and concentration of the analyte
ɛcl = A = log(I0/I) … (1)
where, I0 = the incident light power, I = the transmitted light power, ɛ = molar absorptivity, c
= concentration of analyte and l = thickness of the solution.
(b) Digital-imaging method:
The color and intensity of digital image are usually 24 bit data (8 bit R + 8 bit G + 8 bit B)
forming an additive color space, in which R, G and B lights are added together in various
combinations to reproduce a broad range of colors. By using combination of R, G and B
intensities, many colors can be displayed. The intensity of each color has 256 levels (from 0
to 255). The value of R = 0, G = 0, B = 0 refers to pure black while R = 255, G = 255, B = 255 is
pure white. With this system, unique combinations of R, G and B values are allowed, providing
for millions of different hue, saturation and lightness shades. These extensive dynamic colors
of images provide the database for quantitative analysis. The goal of this study is to employ
digital images-based colorimetry for the determination of Ni2+ concentration in aqueous
samples.
The concentration of analyte is a function of color coordinates: c = f(𝑅𝐺𝐵)… (2)
(ii) Reagents, solutions and Instrumentation: NiSO4 (100 ppm), NaOH (1 N) solution,
Dimethylglyoxime (DMG), K3[Fe(CN)6], Colorimetry and smartphone.
(iii) Reaction Scheme: DMG reacts with Ni2+ to form a pink-colored Ni(dmg)2 complex in
alkaline medium, and gets oxidized by potassium ferricyanide (K3[Fe(CN)6]) to form a brown-
red, water soluble oxidized Ni(dmg)2 complex (Scheme 1). Absorption spectrum of the
oxidized complex shows absorption maxima at a wavelength of 440 nm (Fig. 1). Concentration
of Ni2+ in the given unknown sample is determined from the calibration graph (Fig. 2).

Page | 14
 Scheme - 1

(iv) Procedure:
(a) Colorimetry method:
Take 5 standard 50 mL volumetric flasks (to prepare 4 known and 1 unknown solution).
Fill the burette with Ni stock solution (100 ppm). Add 1, 2, 3 and 4 mL of the Ni solution in
burette to the std. flasks to get 2, 4, 6 and 8 ppm of steel containing nickel(II) solutions. The
unknown sample will be furnished in another 50 ml volumetric flask. Further, add 0.5 mL of
DMG solution followed by 0.5 mL of K3[Fe(CN)6] solution using a burette to all the 5 std. flasks.
All the flasks are shaken well once and waited for 5 minutes. After that, make up the 50 mL
mark in std. flask with 1N NaOH solution. Allow the flasks at least 10 minutes for the complete
complex formation. Absorbance of the formed brown-red solution is measured at 440 nm
against NaOH solution (blank). Record these absorbance readings in Table 1.
Draw a calibration graph taking concentration of Ni2+ (in ppm) as X-axis and absorbance
readings as Y-axis. A straight line that passes through the origin (see Fig. 2) is an indication
that the measured data obeys Beer’s Law. From the calibration plot, measure the
concentration of nickel in the given unknown sample.
(b) Digital imaging method: The prepared standard solutions are lined up along with
unknown concentration sample and blank. Using a white paper as background, take a
photograph of the samples by holding the camera around 50 cm away. Calibration curve will
be constructed through the RGB values of analytical response with different conc. of Ni 2+ ions
using “RGB Tool” APP. In the plotted graph, RGB response varies linearly vs the analyte
concentration. In order to get precise analysis, follow the steps given below:

Page | 15
 RGB Method Flow Chart
RGB Method Flow Chart
RGB Method Flow Chart
Transfer prepared standard solution and unknown solution
Transfer
into prepared
different standard
colorimetric solution
tubes and
testunknown unknown solution
Transfer prepared standard solution and solution
into different colorimetric test tubes
into different colorimetric test tubes
Take image of all test tube solution
Take image of all test tube solution
Takeusing
image smart phone
of all camera
test tube solution
using smart phone camera
using smart phone camera
Open the image processing app (RGB colour detector)
Open the image processing app (RGB colour detector)
Open the image processing app (RGB colour detector)
Go to gallery, open the image stored in app and
Govalues
to gallery,each
openimage/conc.
the image stored in app and
Goextract RGB
to gallery, open the forimage stored in app and
extract RGB values for each image/conc.
extract RGB values for each image/conc.
Process the RGB values (R/G) or (R/B) or (G/B)
etc.,Process thelinear
till values
to get RGB values (R/G) or (R/B) or (G/B)
Process the RGB (R/G)response
or (R/B) or (G/B)
etc., till to get linear response
etc., till to get linear response
Plot the calibration curve using RGB linear
Plot vs
response theconcentration
calibration curve using RGB linear
Plot the calibration curve using RGB linear
response vs concentration
response vs concentration
Find the unknown conc using the
Find the unknown conc using the
Find thecalibration curve
unknown conc using the
calibration curve
calibration curve

Table 1: Experimental Data

S. No. Data collected from
Data collected from smartphone device*
Colorimetric device
Conc. Abs R G B R/G G/B R/B
(ppm, X-axis) (Y- axis)
1.
2.
3.
4.
5. Unknown
*Corresponding ratio that is linearly increasing with analyte concentration is used for plotting
Fig. 2.
Result:
(i) Concentration of Ni in steel sample (using colorimetry) = _________ ppm (mg/L)

Find(ii)
the Concentration
unknown conc.of NiUsing
in steel
thesample (using digital imaging) = _________ ppm (mg/L)
calibration Find the unknown
curve conc. Using the conc. Using the
Find the unknown
calibration curve
calibration curve

Page | 16
 Expt. No.: Date:

Lab scale preparation of important drug intermediate- Synthesis of 2,3-

Experiment
Diphenylquinoxaline

 Preparation of 2,3-Diphenylquinoxaline from benzil

Aim  Calculating the yield of the reaction and confirming the product obtained by
analytical data

Problem
Preparation of a medicinally important organic molecule by condensation reaction
definition

Synthesis of 2,3-Diphenylquinoxalin will be carried out by condensation reaction. After

completion of the reaction, monitored by thin-layer chromatography, the product will be
Solution
filtered and recrystallized. The identification of the product will be carried out by infrared
spectroscopy.

Students will be able to:

Student  monitor the reaction by using thin-layer chromatography and calculate the yield of
learning the reaction
outcomes
 learn about basic organic chemistry transformations and the usage of spectral data
for the identification of the product

Significance of the reaction.

Quinoxaline derivatives are an important class of heterocyclic compounds in pharmaceutical
industries due to their wide range of therapeutic uses and potential activities, such as
antimicrobial agents, cytotoxic agents, anti-tubercular, anxiolytic, anti-HIV, antioxidant, anti-
inflammatory, antimalarial, anticancer, antidepressant, antibacterial, antifungal. Besides, they
are also used in the agricultural field as fungicides, herbicides and insecticides, pesticides. Also,
they are used as dyes, efficient electroluminescent materials, organic semiconductors, corrosion
inhibitor etc. Owing to these plethora of applications, the synthesis of quinoxalines is very
significant in the field of organic chemistry.

Principle.
When benzil or diphenylethanedione (1,2-dicarbonyl compound) is treated with o-
phenylenediamine (1,2-diaminobenzene) in ethanol (solvent), the amine groups and carbonyl
groups get condensed via removal of two water molecules, resulting the formation of 2,3-
diphenylquinoxaline.

Page | 17
Working of thin-layer chromatography (TLC).
Thin Layer Chromatography can be defined as a method of separation or identification of a
mixture of components into individual components by using finely divided adsorbent
solid/(liquid) spread over a plate and liquid as a mobile
phase.
Thin-layer chromatography is performed on a sheet of
glass or plastic, or aluminium foil, which is coated with a
thin layer of adsorbent material, usually silica gel,
aluminium 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.
It is based on the principle of adsorption chromatography or partition chromatography or a
combination of both, depending on the adsorbent, its treatment, and the nature of solvents
employed. The components with more affinity towards the stationary phase travel slower.
Components with less affinity towards the stationary phase travel faster. Once separation
occurs, the individual components are visualized as spots at a respective level of travel on the
plate. Their nature or character is identified through suitable detection techniques.

Chromatography is an important technique that enables the separation, identification, and

purification of the components of a mixture for qualitative and quantitative analysis.
In this physical method of separation, the components to be separated are distributed
between two phases, one of which is stationary (stationary phase) while the other (the mobile
phase) moves in a definite direction. Depending upon the stationary phase and mobile phase
chosen, they can be of different types.

Page | 18
 TLC system consists of:
TLC plates - preferably ready-made with a stationary phase: These are stable and chemically
inert plates, where a thin layer of stationary phase is applied on its whole surface layer. The
stationary phase on the plates is of uniform thickness and is in fine particle size.
TLC chamber - The chamber maintains a uniform environment inside for the proper
development of spots. It also prevents the evaporation of solvents and keeps the process
dust-free.
Mobile phase- This comprises a solvent or solvent mixture. The mobile phase used should be
particulate-free and of the highest purity for the proper development of TLC spots. The
solvents recommended are chemically inert with the sample and the stationary phase.
Applications of Thin Layer Chromatography (TLC):
1. In monitoring the progress of reactions
2. Identify compounds present in a given mixture
Required reagents, apparatus, and conditions:
Reagents and solvents:
Benzil, o-Phenylenediamine, Ethanol
Apparatus:
Beaker, Buchner funnel, Measuring cylinder, Filter paper
Procedure
 To a solution of 2.1 g of benzil in 8 mL of ethanol, add 1.1 g of o-phenylenediamine in 8
mL ethanol in a beaker.
 Warm the reaction mixture in a water bath for 30 min, then add water until a slight
cloudiness persists and allow the reaction mixture to cool.
 Filter the product and dry.
 Recrystallize the product from aqueous ethanol. Filter the recrystallized product and
weigh it.
Table: Calculated amounts of reagents in the stoichiometric reaction

S. No. Compound Weight Molecular weight 𝒘𝒆𝒊𝒈𝒉𝒕 𝒕𝒂𝒌𝒆𝒏

Moles =
𝑴𝒐𝒍𝒆𝒄𝒖𝒍𝒂𝒓 𝒘𝒆𝒊𝒈𝒉𝒕

1 2.1 g

2 1.1 g

Page | 19
Melting Point: 125-126 °C
IR spectral data: Infrared (IR) spectroscopy deals with the infrared region of the electromagnetic
spectrum, that is light with a longer wavelength and lower frequency than visible light. It works
mainly on the absorption of infrared radiation by the molecules/ materials due to the change in
the dipole moment. This vibrational spectroscopy is used to study and identify chemical
substances or functional groups in solid, liquid, or gaseous forms.
IR spectrum of 2,3-diphenylquinoxaline:

Results:

Molecular formula of 2,3-diphenylquinoxaline = C20H14N2

Molecular weight of 2,3-diphenylquinoxaline = …………………

Theoretical yield of 2,3-diphenylquinoxaline = …………………. g

Weight of the product obtained =…………………. g

% Yield = (Weight of the product obtained by the experiment) x 100/(Theoretical Yield)

Page | 20
 Estimation of sulphate ion in drinking water by conductivity method

Sulphate (SO42-) is found in almost all natural water. Origin of most sulphate
compounds is the oxidation of sulphite ores, presence of shales or the industrial wastes.
Ground water moving through soil and rocks containing sulphate minerals result in higher
dissolved sulphate ions than permissible limit.

Problems due to excess sulphate ion concentration in water:

 Sulphates cause scale formation in boilers, pipes, etc.
 High sulphate concentration will leads to corrosion on copper piping.
 Sulphate has a laxative effect and creates diarrhea leading to dehydration in humans
and animals.
 High sulphate concentration leads to eutrophication of water bodies leads to
reduction of dissolved oxygen Sulphate will give bitter taste to water if the
concentration exceeds beyond 250 ppm.

Methods to estimate sulphate ion concentration in water:

1. Turbidimetry method: It involves the measurement of turbidity formed when an

aliquot of BaCl2-gelatin reagent is added to acidified sulphate solution.
2. Titrimetric method: By dissolving precipitated BaSO4 in excess of EDTA solution and
the excess EDTA is back titrated with standard Zinc solution.
3. Colorimetric Measurement: Based on the reaction of barium chloranilate with
sulphate ion at pH 4 in ethanol yield highly colored acid-chloranilate ion and is
measured at 530 nm.
4. Conductometric method: This method measures the conductivity of the solution as
the titration proceeds. Conductance tends to vary with the characteristics of the
solvent, number, size and charge of ions involved. When one ion is replaced by
another ion significantly during the titration, conductance will change in a linear
manner until the replacement is complete. After that, the line will change to different
slope due to the additional inclusion of another ion of difference conductance.

Page | 21
 Expt. No.: Date:
Experiment Estimation of sulphate ion in drinking water by conductivity method
Problem definition People using water with high levels of sulfate are vulnerable to
dehydration and diarrhea. Kids are more sensitive to sulfate than adults.
Methodology Conductivity of the soluble sulphate solution will change when it is
precipitated by BaCl2. Conductivity will reach minima when all sulphate
ions are precipitated, and from which, the total amount of sulphate ion
present in the water can be determined.
Solution Amount of BaCl2 required to remove the dissolved sulphate can be
estimated.
Student learning Students will learn to
outcomes a) perform conductometric method
b) remove sulphate ion from irrigate water

Principle:
Electrolyte solutions conduct electricity due to the presence of ions in solution. In case of
precipitation titration between BaCl2 and Na2SO4, the conductance decreases slowly due to
the replacement of Cl- ion by SO42- ion up to the equivalence point. After the equivalence
point, the conductance increases rapidly due to the excess addition of BaCl2 which remains in
solution as Ba2+ and Cl-. This makes detection of neutralization point easy from the
conductance trend plotted as a graph. This is the principle used in the estimation of SO42-
from contaminated water sample.
Requirements:
Reagents and solutions: BaCl2 (0.1 N), Na2SO4 (0.02 N), unknown sulphate solution and
distilled water.
Apparatus: Conductivity Bridge, Conductivity cell, Burette, Pipette, Volumetric flasks, Glass
rod, Beaker (100 mL).
Procedure:
Calibration of Conductivity meter: Place a freshly prepared 0.1 N KCl solution (given in bottle)
in a 100 mL beaker. Dip the conductivity cell in this solution and connect to the Conductivity
meter. Press “CAL” button and complete the internal calibration of the instrument.

Standardization of BaCl2 (Titration – 1):

Pipette out 20 mL of 0.02 N Na2SO4 solution (from Bottle A) in a 100 mL beaker and add 10
mL of distilled water to it to make the conductivity cell dip completely in the solution. Addition
of water will not affect the conductivity since the number of ions in the solution remains
unaltered. Dip the conductivity cell into the solution in the beaker and connect to the
conductivity meter. Fill the burette with ~0.1 N BaCl2 solution (from Bottle B). Record the
conductivity of the sulphate solution without adding any BaCl2 from the burette (0th reading).
Add 1 mL BaCl2 of known concentration into the beaker, stir with glass rod and note down the

Page | 22
conductance. Continue the addition of BaCl2 (1 mL each time) and note the conductance after
each addition. Continue the titration beyond the equivalence point for about 5 mL. The
conductance will either decrease slightly or remain constant until complete precipitation of
BaSO4, and then starts increasing on continuing the addition of BaCl2. A graph is now drawn
by plotting conductance vs volume of BaCl2 added. Intersection point from the plot gives the
volume of BaCl2 required for precipitating the sulphate present in the known sample.
Estimation of unknown sulphate in the given solution (Titration – 2):
Make up the unknown sulphate solution given in a 100 mL standard flask upto the mark using
distilled water resulting in a solution containing 0.96 mg/mL of sulphate ions (Eq. wt. of SO42-
= 48.03). Pipette out 20 mL of this solution into a 100 mL beaker and add 10 mL distilled water
to it. Dip the conductivity cell and repeat the above procedure with the unknown sulphate
solution to determine the amount of BaCl2 required for precipitating the unknown sulphate
in the sample.
From the two titrations carried out, calculate the amount of sulphate present in the effluent
sample.

Table 1: Conductometric Titrations

Titration-1: Standardization of BaCl2 Titration-2: Estimation of sulphate content
Burette: BaCl2 solution (~0.1 N) Burette: Standardized BaCl2 solution
Beaker: 20 mL of Na2SO4 (0.02 N) + 10 mL Beaker: 20 mL of unknown sulphate solution +
of distilled water 10 mL of distilled water
Conductivity cell, Conductivity meter Conductivity cell, Conductivity meter
Volume of BaCl2 Conductance Volume of BaCl2 added Conductance
added (mL) ( mhos) (mL) ( mhos)
0.0 0.0
1.0 1.0
2.0 2.0
3.0 3.0
4.0 4.0
5.0 5.0
6.0 6.0
7.0 7.0
8.0 8.0
9.0 9.0
10.0 10.0
11.0 11.0
12.0 12.0

Page | 23
Fig 1: Model graphs – 1 and 2 for conductometric estimation of known and unknown sulphate
sample solutions, respectively.

Calculations:
A) Standardization of 0.1 N BaCl2:
(N x V) of BaCl2 solution = (N x V) of sodium sulphate
0.02 N x 20 mL
Strength (in normality) of BaCl2 solution = Volume measured from Plot−1 (V1)
=________N of BaCl2 solution

B).Estimation of unknown sulphate:

(N x V) of irrigation water sample = (N x V) of BaCl2 solution
N of BaCl2 x Volume measured from Plot−2 (V2)
Strength of irrigation water sample = 20 mL
=________N of irrigation water sample
Amount of sulphate present in 1L
= Normality of irrigation water sample x Eq. wt. of SO42- (48.03)
Amount of sulphate present in given sample solution
Strength of irrigation water sample x 48.03 x100
= 1000

=__________ grams in 100 mL

Result: Amount of sulphate in given irrigation water sample =_________ grams.

Page | 24
 Analysis of Iron in an alloy sample by potentiometry

1. Importance of the experiment:

Steel production is an index of national prosperity and economy, and is globally used
in variety of industrial sectors like shipbuilding, automobiles, construction, machinery and
tools. Carbon steel is one variety in which nearly 96% of iron is alloyed with nearly 2% of
carbon and other elements like manganese, chromium, nickel and copper. The composition
is varied for achieving desired strength, ductile and long-term wearing properties. Thus,
qualitative determination of iron in steel is very important.

2. Concept:
Potentiometric titration is a process of determining the quantity of a sample by adding
measured increments of a titrant until the end-point. The potential difference between
indicator and reference electrodes is measured under conditions where the current passed is
sufficiently small to maintain thermodynamic equilibrium. Potentiometric titrations provide
reliable data than conventional titrations with chemical indicators especially with coloured or
turbid solutions. In this experiment, Fe2+ is oxidised to Fe3+ by KMnO4 as a redox titration.

5Fe2+ + MnO4- + 8H+  5Fe3+ + Mn2+ + 4H2O

Change in the concentration of Fe2+ ions during the addition of KMnO4 is monitored by
measuring the solution potential which is the basis for this experiment. From Nernst equation,
a measurable quantity - voltage or potential is related to the concentration of species
(Fe3+/Fe2+) in the solution.
RT Fe3
E  E0  ln( 2  )
nF Fe
At the end point, a rapid change in the potential would be observed indicative of the
complete conversion of Fe2+ to Fe3+. A plot of observed potential vs volume of KMnO4
consumed or its first derivative graph (E/V vs average volume of KMnO4) is used to detect
the titration end point, which in turn, is used to qualitatively measure the amount of Fe 2+.

3. Applications:
Potentiometry method is an electroanalytical technique which can be used to
determine accurately the iron content in steel samples for industrial applications without
using any indicator. This method is also useful for dilute or unknown samples or compositions
for which identification of appropriate chemical indicators are challenging.

Page | 25
 Expt. No.: Date:

Experiment Analysis of Iron in an alloy sample by potentiometry

Mechanical properties of steel depend on its composition. Hence, it is
Problem definition
important to analyze the amount of Iron in steel for industrial applications.
Potentiometric method using KMnO4 (oxidizing agent) to oxidize Fe(II) in
Methodology
steel to Fe(III) facilitates the estimation of Iron in steel.
Solution Estimation of iron (%) in different steel samples.
Students will learn to
Student learning
a) perform potentiometric method
outcomes
b) analyze the composition of iron in different grades of steel

Principle:
Potassium permanganate (KMnO4) oxidizes ferrous ion to ferric ion in the presence of
acid as per the reaction:
5Fe+2 → 5Fe+3 + 5e- .......(1)
MnO4- + 8H+ + 5e- → Mn2+ + 4H2O .......(2)
Overall, 5Fe+2 + MnO4- + 8H+ → 5Fe+3 + Mn2+ + 4H2O

Electrode potential (oxidation potential) in the titration depends upon the concentration of
Fe2+, Fe3+ and H+ ions. To avoid the effect of the change in H+ ion concentration, the titration
is usually carried out in large excess of acid. Oxidation potential of this redox system is given
by
RT Fe3
E  E0  ln( )
nF Fe 2
Connecting the redox electrode (Platinum) with a saturated calomel electrode (SCE)
completes the necessary cell as indicated below:
HgHg2Cl2 (s), Saturated KCl ║ Fe3+, Fe2+Pt
When KMnO4 is added, Fe2+ is oxidized to Fe3+ whose concentration increases with
progressive addition of KMnO4. The observed EMF gradually increases. At the end point, there
will be a sharp increase due to the sudden removal of all Fe 2+ ions. Plot-1: EMF measured (E)
vs Volume of KMnO4 added and Plot-2: E/V vs Average volume of KMnO4 was drawn. End
point of the titration is measured from the Plot-2 graph.

Requirements:
Reagents and solutions: 100 mL of KMnO4 (0.05 N) solution, 100 mL of steel solution, 2 N
H2SO4.

Apparatus: Calomel electrode, Platinum electrode, Potentiometer, Volumetric flasks,

Burette, Pipette, Beakers.

Page | 26
 Fig. 1: Typical apparatus for Fig. 2: Typical Potentiometric titration curves.
potentiometric titrations. (a) Normal curve and (b) First derivative curve.

Procedure:
Calibration of Potentiometer:
Switch on the potentiometer and connect the standard cell terminals to either channel
A (move channel switch to position A) or channel B (move the channel switch to position B).
The meter should read 1.018 V. In case it is not 1.018 V, adjust the std. knob to obtain
reference value.

Estimation of Fe(II) in steel:

Transfer the given unknown steel [containing Fe(II)] solution into a clean 100 mL
standard flask and make the solution up to the mark with distilled water and mix well. Pipette
out 20 mL made up steel sample solution into a clean 100 mL beaker and add one test tube
of dil. H2SO4 (2 N). Place Pt electrode in the beaker and connect to the +ve terminal of the
potentiometer. In another beaker, place 50 mL of saturated KCl solution and dip the SCE in
the solution and connect to the -ve terminal of the potentiometer. Place a salt bridge to
complete the cell. Read the EMF of the cell and note down the value. Add 1 mL of KMnO4
solution from the burette to the beaker containing steel sample solution. Stir the solution
carefully and measure the EMF. Continue the addition of KMnO4 solution and record the EMF
for every 1 mL addition as per procedure till the potential shows a tendency to increase
rapidly. After the abrupt change in cell EMF is observed, continue the titration to take 5 more
reading by adding 1 mL burette solution every time. Plot EMF (ordinate) vs. volume of KMnO4
added (abscissa) to get S-shaped curve which indicate the volume range of the end point.

To find out the volume of end point more precisely, carry out the 2nd titration in similar
way but by adding 1 mL aliquots of KMnO4 initially and then 0.1 mL aliquots between the two
volumes where the end point is detected. Continue the titration beyond the end point as done
above. The exact end point is determined by differential method i.e. by plotting ΔE/ΔV vs
average volume of KMnO4 added. Calculate the normality strength of the Fe(II) in the given
solution.

Page | 27
OBSERVATION AND CALCULATIONS

Potentiometric Titration-I:
Burette: KMnO4 solution (0.05 N)
Beaker: 20 mL of steel solution containing Fe(II) + 20 mL (one test tube) of dil. H2SO4
Electrodes: Indicator electrode (Pt) to red terminal and SCE to black terminal

Volume of Volume of
S. No. EMF (volts) S. No. EMF (volts)
KMnO4 (mL) KMnO4 (mL)
1 11
2 12
3 13
4 14
5 15
6 16
7 17
8 18
9 19
10 20

Fig. 3: Plot of EMF vs Volume of KMnO4 added (mL)

Page | 28
Potentiometric Titration-II:
Burette: KMnO4 solution (0.05 N)
Beaker: 20 mL of steel solution containing Fe(II) + 20 mL (one test tube) of dil. H 2SO4
Electrodes: Indicator electrode (Pt) to red terminal and SCE to black terminal

Sl. Vol. of KMnO4 EMF E/V Average

E (Volt) V (mL)
No. ( mL) (Volt) (Volt/mL) Volume (mL)

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15

Fig. 4: Plot of ΔE/ΔV vs Average volume of KMnO4 added.

Page | 29
Calculation:
(N x V) of steel sample solution = (N x V) of KMnO4
0.05 N x Volume of KMnO4 from Plot−2
N of steel sample solution = 20 mL of steel sample
= _____________N

Amount of Fe present in 1 L of sample solution = Normality of steel sample x At. wt. of Fe (55.85)
Amount of Fe present in given (100 ml) sample solution
𝐍𝐨𝐫𝐦𝐚𝐥𝐢𝐭𝐲 𝐨𝐟 𝐬𝐭𝐞𝐞𝐥 𝐬𝐚𝐦𝐩𝐥𝐞 𝐱 𝟓𝟓.𝟖𝟓 𝐱 𝟏𝟎𝟎
= 𝟏𝟎𝟎𝟎
=________________grams in 100 mL

Result: The amount of Iron present in given steel sample is found to be =_________ grams.

Page | 30
 Expt. No.: Date:

Experiment Preparation of nanosilica by sol- gel method and its characterization

Problem definition Preparation and characterization of nanoparticles of SiO2.

SiO2 nanoparticles are prepared by the sol-gel method and characterized
Methodology
by absorption, diffraction and microscopic techniques
Solution Preparation of SiO2 nanoparticles
Students will learn to
Student learning
a) prepare SiO2 nanoparticles by sol-gel method
outcomes
b) analyze the nanoparticles by instrumental techniques

1. Significance of the experiment:

Nanosilica (SiO2) is a promising material to find applications in various fields such as
drug delivery, gene therapy, and detection of biomolecules, photodynamic therapy and
bioimaging. The salient features are high thermal, chemical and colloidal stability, high surface
area, and good biocompatibility. It was also demonstrated that the immobilization of drugs
on silica nanoparticles can reduce adverse side effects.
SiO2 nanoparticles can be achieved by several methods such as chemical vapour deposition,
plasma and combustion synthesis, hydrothermal and sol-gel methods. Each method has its
merits and demerits. The sol-gel method is one of the promising methods employed in the
preparation of SiO2 nanoparticles.
2. Principle of sol-gel method:
Silicon dioxide (SiO2) nanoparticles were prepared by sol-gel method starting from
tetraethylorthosilicate (Si(OEt)4) precursorReference 1. After dissolving in ethanol, the solution
undergoes hydrolysis with the addition of NaOH pellets and results in very unstable silicon
hydroxide [Si(OH)4]. This turns into a turbid colloidal solution called sol. Sols are dispersions
of colloidal particles (with diameters 1-100 nm) in a solvent. A gel is an inter-connected, rigid
network with pores of sub-micrometer dimensions and polymeric chains whose average
length is greater than a micrometer. The sol-gel method for the preparation of nanosilica
particles is represented in Fig. 1.

Reference 1
A. M. Buckley & M. Greenblatt, Journal of Chemical Education, 71, 599-602 (1994).

Page | 31
Fig. 1. Simplified chart of the sol-gel method for preparation of nanosilica particles (ScCO2 =
supercritical CO2)
Possible chemical transformations:

Further, sol is converted into gel by polycondensation of silicon hydroxide [Si(OH)4] into a
condensed polymeric network (from sol  gel) as shown below.

Page | 32
 On drying, the condensed polymeric network gets converted into a three-dimensional
network result a gel.
Colloidal solution (sol)  Condensed polymeric network (gel)  Dried form (Three-dimensional
network) (gel)

On calcination of the gel, SiO2 nanoparticles are formed. SiO2 nanoparticles are characterized
by powder XRD and Electron Microscopic techniques.
3. Requirements:

Reagents and solutions: Tetraethylorthosilicate (TEOS), ethanol, NaOH pellets and distilled
water.
Apparatus, equipment and Instruments: 50 or 100 mL measuring cylinder, two 100 mL
beakers, glass rod for stirring, funnel and filter papers.
4. Experimental procedure:
In a 100 mL beaker, 5 mL of TEOS is transferred using a measuring cylinder followed
by the addition of 5 mL of ethanol. Then 7.5 mL of distilled water is measured through the
measuring cylinder and is added to the ethanolic solution of TEOS. Later, 2 pellets (exactly) of
NaOH are added by a spatula to the 100 mL beaker containing the reaction mixture. The two
immiscible solutions in the beaker become miscible upon constant mixing them with the glass
rod. The pH of the solution should be basic. This sol-gel solution is left as such under stirring
for 30 min. The sol-gel obtained silica could be visualized in the bottom of the beaker as a
white solid. Later it was filtered using a filter paper.

5. Characterization:
The powder XRD pattern of the collected powder sample was recorded on a Bruker
D8 Advance (Germany) with Cu Kα radiation source (operating Voltage and current 30 KV,
40MA respectively) in the scan range 10-80º (2ϴ). The expected pattern is given in Fig.2 for
reference.
a) Infrared spectrum b) Powder XRD spectrum

Page | 33
c) Scanning electron microscopic image

Fig. 2. a) Infrared spectrum of nanosilica particles; b) XRD pattern of SiO 2 nanoparticles

mediated in ethanol; c) Scanning electron microscopic image of the surface of nanosilica
particles
5. Particle size calculation using Scherrer’s equation:

Scherrer equation:
𝐤∗𝛌
Grain size =
𝐜𝐨𝐬(𝛉)∗(𝐅𝐖𝐇𝐌)
where, k = 0.9, λ = 1.0506 Å, and
“ϴ” and FWHM will be obtained
from powder XRD data.

Results:

(i) Quantity of nanosilica material obtained by the sol-gel method after the filtration
= --------- g
(ii) Particle size calculated as per the “ϴ” given_______________________________.

Page | 34
Determination of hardness of water sample by complexometric titration
before and after ion exchange process

1. Introduction to Hard Water and its Classification:

Water described as “hard” contains high levels of dissolved Ca2+and Mg2+ions. Ground
and surface water dissolve the Ca2+/Mg2+ containing ores/minerals from surrounding soil and
rock and are enriched with these cations. Hardness is most commonly expressed as milligrams
of CaCO3 eq. per liter. Water containing hardness causing species at concentrations below 60
mg/l are generally considered as soft; 60–120 mg/l as moderately hard; 120–180 mg/l, as
hard; and more than 180 mg/l as very hard water. Based on the type of anions (Cl -, SO42-,
HCO3-) associated with Ca2+/Mg2+ ions, the hardness is categorized into temporary
(carbonate, HCO3-) hardness & permanent (non-carbonate, Cl-, SO42-) hardness.
2. Problems caused by Hard Water:
Hard water can cause costly breakdowns in boilers, cooling towers and plumbing.
When hard water is heated, the hardness causing salts tend to precipitate out of solution,
forming a hard scale or soft sludges in pipes and surfaces, thereby completely plugging pipes
and restricting flows. In boilers, scale formation prevents efficient heat transfer, thereby
resulting in energy loss and overheating which could lead to serious accidents. At the
domestic level, hard water lessens the effectiveness of soap by forming scums/precipitates,
which adhere to human skin. Human consumption of water containing excess of Ca and Mg
are associated with increased risks of osteoporosis, nephrolithiasis, colorectal cancer,
hypertension, stroke, coronary artery disease, insulin resistance, diarrhea and obesity.
3. Estimation of Hard Water:
Traditionally, hardness in water is estimated by complexometric titration using sodium
salt of EDTA and EBT as indicator at pH = 9-10. EBT forms an unstable wine-red colored
complex with Ca2+/Mg2+ions, which upon titrating with EDTA, results in the breaking of EBT-
Ca2+/Mg2+ unstable bond and formation of stable EDTA-Ca2+/Mg2+ bond. The endpoint
changes from wine-red (EBT-Ca2+/Mg2+) to steel blue (free EBT).
4. Modern Treatment of Hard Water:
Hard water is made soft by the use of a water softener i.e., ion-exchange resins (IER)
which are very small porous spherical polymeric beads, with specific functional groups
(sulphonic/carboxylic acid) attached to the polymeric backbone. Therefore, the IERs carrying
a negatively charged exchange site can hold a positively charged ion. When the hard water is
passed through the resin beads, Ca2+/Mg2+ions in hard water are exchanged from the solution
for hydrogen/sodium ions, which are much more soluble and does not precipitate out to form
scale or sludges. Eventually, the resin beads get saturated with hardness causing ions and the
exhausted beads are regenerated by using a mild acid or brine solution to flush out the
Ca2+/Mg2+ionsretained in the resin beads.

Page | 35
 Expt. No.: Date:

Determination of hardness of water sample by complexometric

Experiment
titration before and after ion exchange process

Hardness of water is due to the presence of dissolved calcium and

magnesium salts in water. EDTA forms stable complex with hardness
Problem definition
causing salts and is used in the removal of scale and sludge forming
impurities in industrial boilers.
EBT indicator-Metal ion complex is weaker compared to EDTA-metal
Methodology ion complex. The end point is the color change from wine red (EBT-
Metal ion complex) to steel blue (free EBT indicator).
Estimation of Calcium hardness (in ppm) in the given unknown sample.
Solution
Understanding the water softening using ion-exchange resins.
Students will learn to

Student learning a) perform complexometric titration

outcomes b) understand the efficiency of ion-exchange resins using in water
purifiers

Principle:

Ehtylenediaminetetraacetic acid (EDTA) forms complexes with a large number of

cations including Ca2+ and Mg2+ depending upon pH of solution. Hence, it is possible to
determine the total hardness of water using EDTA solution. EDTA in the form of its sodium
salt (H2Y2-) is commonly used in complexometric titration for estimation of metal ion because
pure EDTA (H4Y) is sparingly soluble in water. EDTA has six binding sites (the four carboxylate
groups and the two amino groups) providing six pairs of electrons. The resulting metal-ligand
complex, in which EDTA forms a cage-like structure around the metal ion, is very stable at
specific pH. All metal-EDTA complexes have a 1:1 stoichiometry. The H2Y2- form complexes
with metal ions as follows.

M + H2Y2-  MY2- + 2H+ ----------- (1)

where, M is Ca2+ and Mg2+ present in water. Reaction (1) can be carried out quantitatively at
pH 10 using Eriochrome Black T (EBT) as indicator. EBT forms a wine-red complex with M2+
ions which is relatively less stable than the M2+-EDTA complex. On titration, EDTA first reacts
with free M2+ ions and then with the metal-EBT indicator complex. The latter gives a colour
change from wine-red to steel blue at the equivalence point.

Page | 36
Removal of hardness using ion exchange resins (IER):

Ion exchange is a reversible process. When hard water is passed through cation ion-
exchange resins packed in a narrow column, Ca2+ and Mg+ cations in hard water are
exchanged with Na+ or H+ ions in the resins. The exhausted resins are regenerated by passing
10% dil. HCl through the column. A typical example of application is preparation of high-purity
water for power engineering, electronic and nuclear industries and in household water
purifiers.

Requirements
Reagents and solutions: Standard hard water (1mg/mL of CaCO3 equivalents), 0.01 N EDTA
solution, EBT indicator, hard water sample, NH3-NH4Cl buffer solution and ion exchange resin.
Apparatus: Burette, pipette, conical flask, standard flask burette stand and IER column.
Procedure:

Titration-I: Standardization of EDTA

Pipette out 20 mL of the standard hard water containing 1mg/mL of CaCO3 (1000 ppm)
into a clean conical flask. Add one test tube full of ammonia buffer (NH4OH – NH4Cl) solution
to maintain the pH around 10. Add three drops of Eriochrome Black – T (EBT) indicator and
titrate it against the given EDTA solution taken in the burette. The end point is change of
colour from wine red to steel blue. Repeat the titration for concordant titer values. Let ‘V 1’
be the volume of EDTA consumed.

S. Volume of standard hard water Burette reading (mL) Volume of EDTA

No. (mL)
Initial Final (V1, mL)
1
2
3
Concordant titer value

Page | 37
Calculation:

20 mL of given hard water consumes V1 mL of EDTA

20 mg of CaCO3 requires V1 mL of EDTA for complexation
1 mL of EDTA requires = 20/V1 mg CaCO3 for complexation
This relation will be used in other two titrations

Titration-II: Estimation of total hardness of hard water sample

Pipette out 20 mL of the given sample of hard water into a clean conical flask. Add one
test tube full of ammonia buffer (NH4OH – NH4Cl) solution and three drops of Eriochrome
Black–T (EBT) indicator. Titrate this mixture against standardized EDTA solution taken in the
burette. The end point is the change of color from wine red to steel blue. Repeat the titration
for concordant titer value. Let ‘V2’ be the volume of EDTA consumed.

S. Volume of sample hard water (mL) Burette reading (mL) Volume of EDTA
No.
Initial Final (V2, mL)
1
2
3
Concordant titer value

Calculation:

From Titration 1, we have the following relation:

 1 mL of EDTA requires = 20/V1 mg CaCO3 for complexation
From Titration 2, 20 mL of sample hard water consumes = V2 mL of EDTA
= V2 x 20/V1 mg of CaCO3 eq.
V2 x 20 x 1000 V2 x 1000
1000 mL of hard water sample consumes = ppm = ppm
V1 𝐱 𝟐𝟎 V1

Total hardness of the water sample = “X” ppm

Page | 38
Titration-III: Removal of hardness using ion exchange method

Arrange the ion exchange column on to a burette stand and place a clean funnel on
top of the column. Pour the hard water sample (around 40 to 50 mL) remaining after the
completion of Titration – 2 through the funnel and into the ion exchange column. Place a clean
beaker under the column and collect the water passing through the column over a period of
10minutes. Adjust the valve of the column to match the duration of outflow.

From the water collected through the column, pipette out 20 mL into a clean conical flask and
repeat the EDTA titration as carried out above. Note down the volume of EDTA consumed as
‘V3’.

S. Volume of sample hard water (mL) Burette reading (mL) Volume of EDTA
No.
Initial Final (V3, mL)

Concordant titer value

Calculation:

From Titration 1, we have the following relation:

 1 mL of EDTA requires = 20/V1 mg CaCO3 for complexation

From this relation, it can be seen that

20 mL of water sample after softening through the column consumes = V3 mL of EDTA.

= V3 x 20/V1 mg of CaCO3 eq.

 1000 mL of water sample after softening through the column consumes

V3 x 20 x 1000 V3 x 1000
= ppm= ppm
V1 𝐱 𝟐𝟎 V1

 Residual hardness of the water sample = “Y” ppm

Result:

Total hardness of the water sample= ……. (X) ppm

Residual hardness in the water sample= …… (Y) ppm

Hardness removed through the column = ……… (X–Y) ppm

Page | 39
Demo Expt:1

Size dependent color variation of Cu2O nanoparticles by a spectrophotometer

Importance of the experiment:

Nanoscience and nanotechnology play an important role in our daily life. Materials from processors
of personal computers, mobile phones, light emitting diodes to medical equipment, drugs etc. consist
of nanosized parts with unique properties. The electronics material are mainly based on
semiconductor materials such as silicon and gallium arsenide whose properties depend on their
composition, shape and size. This experiment aims at providing an explanation of size-dependent
optical properties of semiconductor compounds.

Optical Properties of Cu2O:

Cu2O absorbs light in wavelength range from UV to orange and its most common color is red.
However, as Cu2O is a semiconductor, its bandgap depends on size, shape, and crystalline structure
of the sample. Below certain limit (less than micrometer), bandgap changes subsequently changing
the color of Cu2O.

Principle:

When a solution with suspension is exposed to light, part of the incident light energy is
dissipated by absorption, reflection and refraction, while the remaining is transmitted. When the
suspension is viewed at right angles to the direction of the incident light, the system appears
opalescent due to the reflection of light from the particles of suspension (Tyndall effect). The reflection
of light is irregular in nature and therefore the term scattered light is used for it. The measurement of
the intensity of scattered light as a function of the suspended particle concentration is done by the
technique known as Turbidimetry or Nephelometry. The intensity of scattered light is measured at
right angles to the direction of the incident light. Light scattering properties changes with respect to
the particle size.

Page | 40
Reaction scheme:

Cu2O particles can be synthesized via redox reactions between sugar and alkaline solution of copper(II)
complex with citrate anions (Benedict’s reagent). Heating the sugar solution with Benedict’s reagent
in alkali medium leads to sugar oxidation to corresponding carboxylic acid salts and reduction of blue
colored copper(II) ions to Cu2O with other color. Higher reactivity leads to formation of bigger Cu2O
particles.

Reagents & Apparatus:

a) Copper sulphate (CuSO4·5H2O) (d) Sodium hydroxide (NaOH)

(b) Sodium carbonate (Na2CO3) (e) Glucose solution
(c) Trisodium citrate dihydrate (Na3C6H5O7·2H2O) (f) Nephelometer spectrophotometer
[Na3Cit]

Procedure:

(a) Preparation of Benedict’s reagent: Dissolve 6.0 g of CuSO4·5H2O in 20 mL of hot water. Prepare
alkaline citric solution by mixing 4.0 g of Na2CO3 and 7.0 g of Na3Cit in 40 mL of water. Add this citrate
solution in hot CuSO4 solution. After that the mixture was diluted with 40 mL of water. [Conc. of stock
solution = 240 mmol w.r.t. CuSO4.]

(b) Preparation of glucose solution: Prepare glucose solution by dissolving 30.0 g glucose in 750 mL
of water. [Conc. of stock solution = 222 mmol]

(c) Preparation of NaOH solution: Prepare 100 mL of 1.0 M NaOH solution.

(d) Preparation of conditioning agent (Glycerol-alcohol solution): Mix 400 mL absolute ethanol to
200mL of glycerol.

(e) Preparation of Cu2O solution:

Page | 41
 1. Take 50 mL volumetric flask and label them 1, 2, 3, 4, 5 (for Table 1) & A, B, C, D (for Table
2)
Table 1:

Sample No. Vol. of Benedict’s Vol. of glucose Conc. of NaOH Observation/Color

reagent (mL) solution (mL) (M)
1 0.5 4.5 0.0 Yellow


2 0.5 4.5 0.0001
3 0.5 4.5 0.001
4 0.5 4.5 0.01
Red
5 0.5 4.5 0.1

Table 2:

Sample No. Vol. of Benedict’s Vol. of glucose Vol. of NaOH 0.01 M

reagent (mL) solution (mL) (mL)
A 0.5 4.5 45
B 1.0 9.0 40
C 1.5 13.5 35
D 2.0 18.0 30

2. Mix certain amounts of Benedict’s reagent in the volumetric flask and dilute the solution with
45 mL of NaOH solution of certain concentration.
3. To each flask add glucose solution and make the volume to 50 mL (with respective NaOH
solution) and mix well.
4. From the flask, take around 10 mL solution in test tubes and place the test tubes with reaction
mixtures into a beaker with boiling water and keep it there for 5 min.
5. After that, transfer the test tubes to the beaker with room temperature water in order to stop
reaction.
6. The fresh samples can be photographed (Table 1) and use for analysis purpose (Table 2).
7. Next, mix equal volume of conditioning agent (glycerol:ethanol – 1:2) to the sample solution
(10mL of analyte + 10 mL of conditioning agent). Afterwards, transfer the samples (A – D) in
cuvette (the sample tube for turbidity analysis that should be very clean inside and outside
and should not contain any scratches). Note the reading on turbidity scale [Turbidity is
measured in NTU (Nephelometric Turbidity Units)].
8. Prepare a standard curve by plotting turbidity (NTU units) vs the volume of NaOH (Refer the
Appendix in presentation file, .ppt). Then calculate the unknown concentration of copper(II)
by back calculation.
Observations:

Page | 42
 Sample Vol. of NaOH (mL) Turbidity (NTU)

Result:

1) The volume of NaOH for the unknown solution _______ mL.

2) The concentration Cu(II) solution is _____ mmol.

Reference:

N. E. Markina, M. V. Pozharov, A. V. Markin. J. Chem. Educ. 2016, 93, 4, 704–707.

Page | 43
Demo Expt:2
Computational Chemistry: Geometry optimization using Avogadro software
Aim
To create models and optimize the geometry of H2O and H2S dimers, and 2-nitrophenol as
examples of inter- and intra-molecular hydrogen bonding respectively using Avogadro
software.
Course outcome
Using the Avogadro software students will learn about modelling 3D structures of any
molecule of interest. Also, they will learn the concept of geometry optimization and
associated stabilization parameters like hydrogen bonding interaction.
Geometry optimization
The geometry optimization can be performed employing commonly used methods like
steepest descent and conjugate gradient methods that are available in Avogadro software to
calculate the energy of given molecular geometry along with the application of force field or
molecular mechanics theory.
The force field is represented as a collection of equations and associated constants that are
designed with standard parameters to reproduce geometry and interpret the behaviour of
atoms and molecules [Ref: https://doi.org/10.1016/j.colsurfb.2014.06.050].

Modelling procedure
Avogadro is open source software which is available to model chemical structures and predict
their approximate energy values. Avogadro software can be downloaded from the
https://avogadro.cc/

Representative examples of molecular systems considered in this demonstration are:-

1. Comparison and modelling of water (H2O) dimer and H2S dimer to investigate the
influence of intermolecular hydrogen bonding.

Page | 44
2. Modelling of 2-nitrophenol to study the influence of intramolecular hydrogen bonding.

These examples will provide clear understanding on the concept of hydrogen bonding that
stabilizes the molecular systems.
Step 1: Make sure that in “Settings” menu, the options “Display types”, “Draw settings” are
enabled as shown below. In the “Display types” enable “Hydrogen bond” to visualize it in the
optimized structure.

Step 2: In the “Draw settings”, choose oxygen atom to model two water molecules placed
close to each other as shown below.

Page | 45
Step 3: In the “Extensions” main menu, choose “Molecular Mechanics” followed by “Setup
force field” menu as shown below.

Once it is chosen, the following screen appears with the option to choose the type of method.

It is possible to perform geometry optimization after choosing the force field and algorithm,
Step 4: Click “Optimize Geometry” in the “Extension” menu to perform geometry
optimization.

Page | 46
Step 5: Now the geometry optimization is performed for water dimer. The optimized
geometry is displayed in the screen. One can visualize the dotted line which indicates the
hydrogen bond between the oxygen atom of one water molecule to the hydrogen atom of
another water molecule. This is an example of intermolecular hydrogen bonding as it involves
two different molecules. Note the energy of the water dimer.

Step 6: Now in the build option, choose “sulphur” to convert water dimer to H 2S dimer.
Remove “Adjust Hydrogens” option in the build menu and click oxygen atoms. Now the
structure becomes H2S dimer as shown below.

Page | 47
Step 7: Now perform geometry optimization as given in Step 4

Note the difference in structure of H2S dimer and the energy.

Question: What brings the change in bond length and bond angle between the two dimers,
H2O and H2S? Explain why hydrogen bonding is missing in H2S dimer?
Answer: The interactions between the H2S molecules are comparatively weaker (more like
van der Waals interaction) than the interaction between the two H2O molecules. That is the
reason the hydrogen bond is absent in H2S dimer. For the same reason, H2O is liquid and H2S
is gas at room temperature.
Note: The software is programmed in such a way that depending on the geometry criteria of
the molecules, the presence of hydrogen bonding will be highlighted.

Page | 48
Step 8: Draw 2-nitrophenol as shown below

Step 9: Perform geometry optimization of 2-nitrophenol as given in Step 4. Once geometry

optimization is performed, intramolecular hydrogen bonding between -OH and -NO2 groups
can be visualized as a dotted line as shown below.

Results:
1. Visualize the geometry of H2O and H2S dimers.
2. The energy of H2O dimer = _____________ and H2S dimer = _____________
3. Visualize the geometry of 2-nitrophenol.
4. The energy of 2-nitrophenol = _____________

Happy Learning!

Page | 49