Challenging experiments lab manual

S.No Experiment Page

1 Comprehensive Water Analysis – Environmental analyses 1
2 Determination of the Calorific Value of an energy source (fuels or food 10
products) by Bomb Calorimeter – Energy (Chemical and bio-)
source evaluation
3 Removal of Heavy Metals from Wastewater using HAP nanoparticles 13
prepared from Egg Shell Bio-Waste – Waste to Wealth
4 Determination of specific rotation by polarimetry- Kinetics of inversion of 19
sucrose in normal sugar and refined sugar samples – Food Quality
5 Building Voltaic Cells or a stable Battery from Waste Metal – 24
Sustainability Battery construction
6 Building Dye Sensitized Solar Cells as Renewable Resource from 27
Natural Dye – Solar cells – Energy source construction

7 Fabrication and Evaluation of a Supercapacitor from the Bio-waste 29

Derived from Sweet Lime Peels - Energy source construction
 Experiment 1: Comprehensive Water Analysis
Aim:
To analyze hardness, chloride content, Total Dissolved Solids (TDS), pH, and sulfate content
of a locally available water sample and compare the water quality as per central pollution
control board (CPCB), India.
Part I - Estimation of Total Hardness of a given water sample
Aim: To determine the total hardness of the given water sample
Principle:
Hardness of a ground water sample is due to the presence of calcium and magnesium salts
present in it. There are two types of water hardness, namely temporary and permanent
hardness. Temporary hardness causing salts, Ca(HCO3)2 and Mg(HCO3)2 (bicarbonates of
calcium and magnesium) which can be removed by simple boiling. On boiling, these salts
form precipitates of CaCO3 and Mg(OH)2 which can be further removed by filtration.
Permanent hardness is caused by the presence of chlorides and sulphates of calcium and
magnesium. Ethylenediaminetetraacetic acid (EDTA) forms complexes with many cations
including Ca2+ and Mg2+ at a basic pH (9-10). Basic pH of the water sample is maintained by
using ammonia buffer (NH4OH – NH4Cl). Hence, it is possible to determine the total hardness
of water sample using EDTA solution. As EDTA is insoluble in water, disodium salt of EDTA is
generally used for the complexation. EDTA has six binding sites (the four carboxylate groups
and the two amino groups, Fig. 1a.) providing six pairs of electrons.

(a) (b)
Structure of EDTA and EBT
To indicate the end of complexation, Eriochrome Black–T (EBT) indicator (Fig. 1b.) is used.
Ca2+ and Mg2+ ions form a wine red colored complex with EBT indicator in the presence of
ammonia buffer solution. When EDTA solution is added to the indicator complex, EDTA
replaces the indicator and a stable complex of Metal–EDTA is formed. The color of the
solution will be steel blue due to the release of EBT indicator into the solution.

Ca2+/Mg2+ + EBT → [Ca2+/Mg2+–EBT]

in water wine red color
(unstable)
2+ 2+
[Ca /Mg –EBT] + EDTA → [Ca2+/Mg2+–EDTA] + EBT
wine red color colorless steel blue color
(stable)

1
 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.
Apparatus: Burette, pipette, conical flask, standard flask, and burette stand.

Procedure:
Titration-I: Standardization of EDTA
20 mL of the standard hard water containing 1mg/mL of CaCO3 (1000 ppm) is pipetted out
into a clean conical flask followed by 3 to 5 ml of of ammonia buffer (NH4OH – NH4Cl)
solution to maintain the pH around 10. Three drops of Eriochrome Black – T (EBT) indicator
was added and titrated against the given EDTA solution taken in the burette. The end point
is change of colour from wine red to steel blue. Titrations are repeated to get concordant
titre value. (V1).
Titration-I: Standardization of EDTA
Standard Hardwater vs. EDTA

S. No. Volume of standard Burette reading Volume of

hard water (mL) (mL) EDTA (V1,
Initial Final mL)

Concordant titre value

Calculation:

20 mL of given hard water consumes V1 mL of EDTA

20 mg of standard hard water requires V1 mL of EDTA for complexation

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

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

2
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 titre value. Let ‘V2’ be the volume
of EDTA consumed.

Titration-II: Estimation of Total hardness of water sample

Standard EDTA vs. given water sample

S. No. Volume of Burette reading Volume of
sample hard Initial Final EDTA (V2,
water (mL) mL)
1

Concordant titre value

From Titration 1, we have the following relation:

Therefore, 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.

Therefore, 1000 mL of hard water sample consumes = V2 x 20/V1×1000/20

= V2/V1×1000 ppm

Therefore, the Total hardness of the water sample = ppm

Result: Total hardness of the water sample = ppm

Part II - Estimation of Chloride content

Solutions of electrolyte conduct electricity due to the presence of ions. In case of conductometric
precipitation titration between AgNO3 and Chloride ions, the conductance decreases slowly due to the
removal of chloride ions as AgCl precipitate preceded by replacement of sodium ions by silver ions up to

3
the equivalence point. After the equivalence point, where no chloride ions are present, the conductance
increases rapidly due to silver ions from the added AgNO3 solution.
Cl-(aq) + Ag+(aq) + NO3-(aq) → AgCl(s)↓ + NO3-(aq)
The obtained precipitate has a low solubility in water. In addition, because only halide and sulfide ions
precipitate in presence of AgNO3, such methods have a good selectivity in quantitative analysis of
chloride ions. The representative titration graph is given below.
Reagents and solutions: Standard AgNO3, and NaCl solutions
Apparatus: Burette, pipette, beaker, standard flask, burette stand, Pt electrode, a digital conductometer
and magnetic stir bar
Procedure:
The given unknown chloride solution is made up to the mark in the 100 mL standard measuring flask
using distilled water. 20 mL of this solution is pipetted out into a 100 mL beaker and 10 mL distilled water
is added to it. A conductivity cell of known cell constant is dipped in it and connected to the conductivity
meter. The burette is then filled with ~0.01 N AgNO3 solution (Bottle B). The conductivity of the unknown
chloride sample without adding any AgNO3 solution from the burette (0th reading) is noted followed by 1
mL AgNO3 solution of known concentration into the beaker with continuous stirring by magnetic stirrer.
Addition of AgNO3 solution (1 mL each time) is continued and the conductance is measured after each
addition. The titration is continued beyond the equivalence point for about 5 mL A graph is now drawn by
plotting conductance vs volume of AgNO3 added. Intersection point from the plot gives the volume of
AgNO3 required for precipitating the chloride present in the water sample.
Model graph

4
 Conductance measurement: Volume of AgNO3 added vs. Electrical Conductivity

Tabulation:
Volume of
S.No. Conductance (mhos)
AgNO3 (mL)

Calculations:
Volume of AgNO3 (from the graph) V1 = mL Normality of AgNO3 N1 = 0.01 N
Volume of Water sample V2 = 20 mL
Normality of given water sample N2 = 𝑉1 𝑁1/𝑉2
Amount of chloride present in 1L = Normality of given water sample x Equivalent weight of
Cl- (35.453)

𝑆𝑡𝑟𝑒𝑛𝑔𝑡ℎ 𝑜𝑓 𝑤𝑎𝑡𝑒𝑟 𝑠𝑎𝑚𝑝𝑙𝑒 𝑥 35.453 𝑥100

Amount of chloride present in 100 ml of the sample solution =
1000

= g

Result: Amount of chloride in given water sample in 100 ml = g

5
 Part III - Estimation of Total Dissolved Solids (TDS) estimation

Aim: To determine the total dissolved solids of the given water sample
Principle: The use of electrical conductivity (EC) is based on the observed linear relationship between
specific conductance and ionic strength for fluids with conductance ranging from 10– 10,000 μS/cm. A
common expression is:
𝑇𝐷𝑆 (𝑚𝑔 ⁄ 𝐿) = 𝑘𝑒 × 𝐸𝐶(𝜇𝑆 ⁄ 𝑐𝑚)
where ke is a constant of proportionality. The TDS-EC relationship is affected by the ionic composition
of the water and the concentration of dissolved species. For most natural waters, the ke value varies
from 0.55 to 0.85, and the average of these two extremes (0.7) is widely used.
Apparatus: Conductivity meter, conductivity cell
Procedure:
The conductivity of the water sample is recorded. The recorded conductivity when multiplied by 0.7
(ke) as per the equation, gives the total dissolved solids present in the given water sample.
𝑇𝐷𝑆 (𝑚𝑔 ⁄ 𝐿) = 𝑘𝑒 × 𝐸𝐶(𝜇𝑆 ⁄ 𝑐𝑚)
Result: Total Dissolved Solids of the given water sample = mg/L

Part IV - Determination of pH of the given water sample

Aim: To determine the pH of the given water sample
Principle:
The term “pH” refers to the measurement of hydrogen ion activity in the solution. Since the direct
measurement of the pH is very difficult, specific electrodes are needed for quick and accurate pH
determination. pH is measured on a scale of 0 to 14, with lower values indicating high H+ (more acidic)
and higher values indicating low H+ ion activity (less acidic). A pH of 7 is considered as neutral. Every
whole unit in pH represents a ten-fold increase in or decrease in hydrogen ion concentration. Most
natural waters possess the pH values ranging from 5.0 to 8.5. Rain water have a pH value of 5.4 to 6.0
which then reacts with the soils and minerals causing the reduction in H+ ion concentration and thus
the water may become alkaline with a pH 0f 8.0-8.5. More acid water (pH<5) and more alkaline (pH
>9) and other immediate changes in the hydrogen ion concentration (pH) suggest that the quality of
the water is adversely affected due to the introduction of some toxic contaminants in water bodies. pH
is measured using pH meter, which comprises a detecting unit consisting of a glass electrode,
reference electrode, usually a calomel electrode connected by KCl Bridge to the pH sensitive glass
electrode and an indicating unit which indicates the pH corresponding to the electromotive force is
then detected. Before measurement, pH meter should be calibrated by using at least two buffers.
Procedure:
The pH electrode is just dipped in a beaker of sample water and the displayed pH value is recorded. To
6
avoid any kind of contamination a clean and dry beaker was used before pouring the sample water.
Result: pH of the given water sample =

Part V - Estimation of Sulfate in the given water sample

Aim: To estimate the amount of sulphate present in the given water sample
Principle:
To estimate sulfate ions, it is converted to a barium sulfate suspension under controlled conditions.
The resulting turbidity is determined by a nephelometer and compared with a curve prepared from
standard sulfate solution.
(Reference: https://www.epa.gov/sites/default/files/2015-12/documents/9038.pdf)

𝐵𝑎𝐶𝑙2 + 𝑆𝑂42− = 𝐵𝑎𝑆𝑂4 ↓ +2𝐶𝑙 −

Schematic diagram of Light scattering in Nephelometry

Reagents and solutions: Conc. HCl, conditioning reagent, Na2SO4 stock solution (1420 ppm) and
BaCl2.
Conditioning reagent: It is prepared by mixing 30 mL of concentrated HCl dissolved in 300 mL of
distilled water, 95% ethanol or isopropanol, and 75 g NaCl in a container. Further, to this mixture, 50
ml of glycerol is added and used for the analysis.
Apparatus: Magnetic stirrer, Nephelometer, Stopwatch, Measuring spoon, 250 mL conical flask, and
1L volumetric flask.
Procedure:
Seven 100 ml standard measuring flasks are taken and labelled as 1, 2, 3, 4, 5, 6 and 7. Flask labeled

7
as 1 is used for the blank solution preparation and 7 is used for the given unknown sample. Standard
Na2SO4 solutions of concentrations, 50, 100, 150, 200 and 250 ppm are prepared by diluting 3.5, 7,
10.5, 14, 17.5 ml of the stock solution of Na2SO4 using distilled water and made up to 100 ml in the
standard measuring flasks and are labelled as 2,3,4,5, and 6. Different concentrations of sulphate
solutions each as prepared are transferred completely to 250-mL Erlenmeyer flasks (conical flasks),
including the blank. To the blank solution in the conical flask, 5 ml of conditioning reagent is added
and stirred in the orbital shaker exactly for 1 min and 0.6 g of BaCl2 crystals alone are added while
stirring, but without sulphate solution. The blank solution is then transferred to the sample tube and
turbidity value is adjusted to zero using the turbidity meter. After calibrating the turbidity values using
the blank solution, turbidity values are to be found for the standard solutions. The made-up standard
solutions in the standard measuring flasks are transferred to the conical flasks. To the contents in the
conical flasks, exactly 5.0 mL of conditioning reagent is added to the sulphate solutions and stirred for
one minute using the orbital shaker. While the solution is being stirred, 0.6g of BaCl₂ crystals is added.
After stirring, each sample was transferred to the sample tube. Readings on the turbidity scale
(Nephelometric turbidity unit, NTU) is noted for all standard solutions one by one. After measuring the
NTU values for all the standard solutions, given unknown solution is also analyzed using the same way.
A calibration plot (straight line) is plotted using the NTU values for the standard solutions against their
concentrations. Concentration of unknown solution was found by extrapolating the NTU value in ‘y’
axis (turbidity, NTU) to meet the straight line and extrapolated to the ‘x’ axis (shown as ‘X’ in the plot).

Model plot of sulphate concentration range vs. turbidity values

8
 Table:
S.No. Concentration (ppm) Turbidity (NTU)

1. 50

2 100

3. 150

4. 200

5. 250

6. Unknown

Comprehensive Report of Water Sample Analysis

Sample Collected from
Date of Collection
Recommended Values for
Recorded Deviation from the
Parameters drinking water as per BIS
Values standard (%)
(Acceptable limit)
Hardness (ppm) 200
Chloride content (ppm) 250
TDS (ppm) 500
pH 6.5 – 8.5
Sulfate content (ppm) 200

References (weblinks):
Water quality BIS standards - https://cpcb.nic.in/wqstandards/
TDS Estimation - https://pure.psu.edu/en/publications/relationship-between-total-dissolved-solids-and-
electrical-conduc or https://doi.org/10.2166/wst.2018.092)
pH measurement: (Reference: https://nitsri.ac.in/Department/Civil%20Engineering/CWE-
301_WATER_QUALITY_AND_ENVIRONMENT_pH_Meters.pdf)

*****

9
Experiment 2: Determination of the Calorific Value of
an energy source (fuels or food products) by Bomb
Calorimeter
Aim:
Collect a solid fuel like coal, or biomass and food product like dried popcorn, grains,
packaged food or dry fruits. A stipulated amount of fuel and food to be controlled burnt under
oxygen atmosphere to estimate Calorific value by use of Bomb Calorimeter.

Principle:
A known mass of the sample (solid) is burnt completely in excess of oxygen. Surrounding
water of the combustion chamber and calorimeter (water equivalent) absorbs the liberated
heat. Thus the heat liberated during the combustion of fuel is equal to the heat absorbed by
water and copper calorimeter. The higher calorific value (HCV) or gross calorific value (GCV)
of the fuel is calculated from the data. As the bomb calorimeter consists of a stainless steel
airtight sealed cylindrical bomb which has an inlet valve for oxygen and is provided with two
stainless steel electrodes. To one of the electrodes, a small ring attached where a nickel or
stainless steel crucible can be supported. The bomb is placed in a copper calorimeter, which
is surrounded by an airtight and water jacket to prevent heat loss due to radiation. The
calorimeter is provided with an electrically operated stirrer and Beckman’s thermometer.

Working formula:
Weight of the fuel= m g
Weight of water taken in the calorimeter = W1 g
Water equivalent of calorimeter = W2 g
Initial temperature of water = t1 °C
Final temperature of water = t2 °C
Specific heat of water = 4.187 kJ/kg°C

10
 (𝑊1 +𝑊2 )×(𝑡2 −𝑡1 )×4.187
𝐺𝐶𝑉 = 𝑚
𝑘𝐽/𝑘 ………………….. (1)

Requirements:
Reagents and solutions: Oxygen Cylinder, Benzoic Acid, Solid Fuel, and food products

Apparatus: Bomb Calorimeter, spatula and butter paper

Procedures:
Calibration of bomb calorimeter is done with respect to standard Benzoic acid which has the
GCV = 26455 kJ/g. To calibrate a known mass of the benzoic acid is taken in clean crucible
that is then supported over the ring while a fine magnesium wire touching the fuel sample is
then stretched across the electrodes. The bomb lid is tightly screwed and filled with oxygen
to 25atmosphere. The bomb is then placed into copper calorimeter, containing a known mass
of water. The stirrer is worked and initial temperature of the water is noted. The electrodes
are then connected to 6volt battery and circuit is completed. The sample burns and heat is
liberated. Stirring of water is continued and the maximum temperature attained is recorded.
After getting the required parameters equation (1) is used to calibrate the instrument by
declaring water equivalent of calorimeter (W2 g).

Table I: Calibration or the estimation of equivalent water of Bomb Calorimeter

Parameters Recorded Values Calculated Value

mB=Mass of Benzoic Acid (g)
Water Taken in the Calorimeter (g)
Initial Temperature t1 (degree
Celsius)
Final Temperature t2 (degree
Celsius)
Water equivalent of Calorimeter,
Nil
W2(g)

Calculation:
26455 × 𝑚𝐵
𝑊2 = − 𝑊1
(𝑡2 − 𝑡1 ) × 4.187

To measure the calorific value of a sample, a known mass of it is placed in clean crucible that
is then supported over the ring while a fine magnesium wire touching the fuel sample is then
stretched across the electrodes. The similar process like calibration has to be followed to
record the initial and final temperature. Then using the equation (1) GCV is estimated.

Table II: Estimation of Fuel Calorific Value

Parameters Recorded Values

mfuel=Mass of Solid Fuel (g)
Water Taken in the Calorimeter (g)
Initial Temperature t3 (degree Celsius)
Final Temperature t4 (degree Celsius)
Water equivalent of Calorimeter, W2(g)

11
 (𝑊1 + 𝑊2 ) × (𝑡4 − 𝑡3 ) × 4.187
𝐺𝐶𝑉𝑓𝑢𝑒𝑙 = 𝑘𝐽/𝑘𝑔
𝑚𝑓𝑢𝑒𝑙

=…………………kJ/kg

OR, Table III: Estimation of Food Calorific Value

Parameters Recorded Values

mfood=Mass of Solid Dry Food (g)
Water Taken in the Calorimeter (g)
Initial Temperature t5 (degree Celsius)
Final Temperature t6 (degree Celsius)
Water equivalent of Calorimeter, W2(g)

(𝑊1 + 𝑊2 ) × (𝑡6 − 𝑡5 ) × 4.187

𝐺𝐶𝑉𝑓𝑜𝑜𝑑 = 𝑘𝐽/𝑘𝑔
𝑚𝑓𝑜𝑜𝑑

=…………………kJ/kg

Result: The estimated calorific value of food or fuel is ………………. kJ/kg

12
Experiment 3: Removal of Heavy Metals from
Wastewater using HAP nanoparticles prepared from
Egg Shell Bio-Waste
Aim:

The objective of this experiment is to produce hydroxyapatite nanoparticles from eggshell

bio-waste by chemical precipitation method and to investigate their potential use in the
removal of Ni (II) from wastewater.

Principles:
Hydroxyapatite (HAP) nanoparticles find application in various fields such as bio-ceramics,
medicine, the pharmaceutical industry, and environmental pollution control. Hence, it is of
great importance to synthesize HAP nanoparticles. HAP nanoparticles from eggshell bio-
waste were prepared by chemical precipitation method and characterized by powder X-ray
diffraction (powder XRD). Hydroxyapatite (HAP), is chemically represented as
Ca10(PO4)6(OH)2. The Ca/P ratio of stoichiometric hydroxyapatite is usually 1.67, whereas
the Ca/P ratio of non-stoichiometric calcium deficient hydroxyapatite is below 1.67.
Compared to synthetic HAP, natural HAP is non-stoichiometric because it contains trace
amounts3 of Na+, Zn2+, Mg2+, K+, Sr2+, Ba2+, F−, and CO32−. Calcium-deficient non-
stoichiometric HAPs derived from natural sources have been shown to be better adsorbents
for the purification of aqueous solutions containing metal cations, anions, and organic
pollutants because of the presence of exchangeable cations, hydroxyl ions, and phosphate
chelating groups. HAP can be synthesized from both chemical and natural sources. Currently,
studies have focused on the use of bio-waste sources, such as coral, egg shells, fish bones,
and seashells (Fig.1), for the preparation of HAP. Among the above sources, eggshell waste
is considered a potential material because of its large availability and rich calcium content (>
95wt% CaCO3).

Fig.1: Biowaste sources available for HAP synthesis

13
 Part A
(i) Synthesis of HAP nanoparticles from Eggshell Biowaste
Requirements:
Cleaned the eggshell
Concentrated HNO3
NaOH
(NH4)2HPO4

Procedures:
Hydroxyapatite nanoparticles are prepared from eggshell bio-waste by simple chemical
precipitation method. First, clean the eggshell biowaste with tap water several times followed
by distilled water to remove dirt and organic residues from the eggshell and then dry it in hot
air oven at 100◦C. Crush the cleaned and dried eggshell bio-waste using agate mortar to get
a fine powder of eggshell. Weigh around 1g of powdered eggshell bio-waste and transfer
into a 100 ml beaker and then add 5 mL of concentrated HNO3 slowly. Stir the mixture
continuously until the eggshell powder dissolves completely in nitric acid. To this, add 15 ml
of deionized water followed by 15 ml of 4N NaOH solution to get yellow coloured turbid
solution. The resultant yellow turbid solution is passed through filter paper to get clear
aqueous solution of Ca(NO3)2. A white milky precipitate of amorphous hydroxyapatite are
obtained by adding slowly 10 ml of (NH4)2HPO4 (0.1 M) to Ca(NO3)2 solution (pH of the
solution maintained at 9). The precipitate is then filtered through Whatman filter paper and
washed several times with deionized water to remove any excess untreated precursors and
byproducts (NH4+ and NO3− ions). Finally, the precipitate is allowed to dry for 1 h at 100 °C
in a hot air oven followed by annealing in an air atmosphere at 900 °C for 3 h using an
electrical furnace to get crystalline hydroxyapatite [Ca10(PO4)6(OH)2] (Scheme 1).

Scheme:

Scheme 1: Synthesis of HAP nanoparticle from egg-shell bio-waste

Characterization:

The powder XRD pattern of the collected powder sample is recorded on a Rigaku Smartlab 3
KW X-ray diffractometer with Cu K radiation source (λ=1.54060 Å) in the scan range 10-
80° (2) and results are compared with standard XRD pattern of HAP is given in Fig.2.

14
 JCPDS 00-009-0432

(211)
(112)
(300)
(002)

(123)
(222)
(202)

(004)
(310)
(210)

(312)

(304)
20 25 30 35 (311)
40 45 50 55 60 65 70
2
Fig.2. Standard XRD pattern (JCPDS file #09-0432) of HAP nanoparticles

Calculating the crystallite size of synthesized HAP nanoparticle using Scherrer’s equation:

Scherrer equation

Grain size = k*  / cos(θ)*(FWHM)

k = 0.9,  = 1.0506 Å, and “” and FWHM to be obtained from powder XRD data.

Fig.3: Calculation of FWHM from XRD data of HAP

Use the following powder XRD data of HAP nanoparticles for calculating the crystallite size

2 Intensity
30.5 13.5
30.6 17.5
30.6 24.3
30.7 22.3
15
 30.7 24.7
30.8 24.1
30.8 24.0
30.9 27.5
30.9 25.3
31.0 27.6
31.0 32.4
31.1 30.4
31.1 27.4
31.2 30.5
31.2 30.1
31.3 43.9
31.3 78.8
31.4 165.7
31.4 262.2
31.5 393.1
31.5 359.3
31.6 237.0
31.6 121.3
31.7 60.1
31.7 49.5
31.8 34.7
31.8 27.0
31.9 26.8
31.9 32.0

Part B

(i) Procedure for estimation of Ni2+ using dimethylglyoxime (DMG) reagent by colorimetry

Principle: Dimethylglyoxime (DMG) reacts with nickel ions and forms a pink-colored Ni(DMG)2 complex
in alkaline medium. It gets oxidized by potassium ferricyanide (K3[Fe(CN)6]) in alkaline medium to form
a brown-red, water soluble oxidized Ni(DMG)2 complex as shown in the Scheme 2. Absorption
spectrum of the oxidized complex shows absorption maxima at a wavelength of 440 nm (Fig.4).

Scheme 2: Formation of brown-red, water soluble oxidized Ni(DMG)2 complex

16
 Requirements:
Reagents and solutions: NiSO4 (1000 ppm), NaOH (1 N) solution, DMG (2 % in 2 N NaOH),
K3[Fe(CN)6] (1.5 %).
Apparatus: Colorimetry, Volumetric flasks, Beakers.
Procedure: Take 6 standard 50 mL volumetric flasks (including the unknown). To every flask add 0.5
mL of DMG solution followed by 0.5 mL of K3[Fe(CN)6] solution using a burette. Further, add 0.5, 1,
1.5, 2 and 2.5 mL of the working Ni stock solution (1000 ppm) from a burette to get 10, 20, 30, 40
and 50 ppm of steel containing nickel (II) solutions. All the flasks are shaken well and made up to 50
mL mark with 1N NaOH solution. Allow the flasks at least 15 minutes after adding all the reagents for
the complete complex formation. Then the absorbance of the formed brown-red solution is measured
at 440 nm against distilled water (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 (Fig.5) 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.

(ii) Removal of Ni2+ ions from wastewater using HAP nanoparticles

HAP can be employed to effectively remove Nickel (Ni2+) ions from wastewater in batch adsorption
studies. First, HAP (0.1 g) was dispersed in 30 ml of deionized water and added to 10 mL of
wastewater containing Ni2+ ions. The mixture was shaken at 180 rpm for 1 h at room temperature in
an orbital shaker. Subsequently, the HAP was filtered using Whatman filter paper, and the residual
Ni2+ content in the filtrate was estimated using a dimethylglyoxime (DMG) colorimetric reagent at a
wavelength of 440 nm detailed above. Finally, the percentage of Ni2+ ions adsorbed on HAP
nanoparticles was calculated based on the initial and final concentrations of Ni2+ ions in the wastewater
using the following equation:
(𝐶0 − 𝐶𝑒 ) × 100
% 𝑅𝑒𝑚𝑜𝑣𝑎𝑙 =
𝐶0

Fig.4: Absorption spectrum of oxidized Ni(II)-DMG

Fig.5: Model calibration curve for Ni(II)
complex showing λmax at 440 nm
determination

Table 1: Experimental Data:

S. No. Concentration (ppm) (X- Absorbance (Y-axis)

axis)
1.
2.
3.

17
 4.
5.
6. Unknown

Results:
(i) The powder XRD peak positions for the prepared HAP nanoparticles are
_______________________________________

(ii) Crystallite size of HAP is _________

(iii) The percentage removal of Ni2+ ions from wastewater is ________

18
Experiment 4: Kinetics of breakdown of sugar from
food samples
Aim:
To follow the kinetics of breakdown of sugar in food samples by following the acid catalysed
hydrolysis reaction.

Principle:
When an ordinary light passes through a nicol prism which is made up of iceland spas, the
emergent light is plane polarised. That is, vibration takes place in one plane. When a
second Nicol prism is placed at right angles to the first, no light can pass through and the
field view appears dark. On rotating the second prism, the field view appears alternately
bright and dark. The maximum darkness follows the minimum as the prism is rotated
through an angle of 90. The first prism is polarizer and the second prism is analyser.

Suppose the axes of the two prisms are at right angles to each other so that the field of view
appears dark. At this position, if a tube containing a solution of cane sugar is placed
between the two prisms, the field lights up. The analyser must be turned through a certain
angle and to make the field again dark. Therefore the solution of cane sugar has the power
of rotating the plane of polarised light through a certain angle and is called optically active
substance.

With optically active substances, if the analyzer needs to be turned clockwise to obtain
darkness, it is ‘dextro rotatory’ of the turn is anti-clockwise, it is ‘laevorotatory’.

The angle of rotation depends upon

a) Nature of substance

b) Length of tube containing solution through which light passes

c) Nature of solvent

d) Temperature

e) Wavelength of light used

If the solution is produced by a liquid which contain 1g of active substance per one ml, when
the light passes through 10cm, it is called ‘specific rotation ‘[]’.

 
l.d

Where l = length of column of liquid in decimeter and

d = density

Taking into account the factors affecting angle of rotation.

19
  100
l.c

 100d
l. p.d
Where C = number of grams of the active substance in 100ml of the solution and p is the
number of grams of the active substance in 100gram of solution.

The Polarimeter

F E D
o C B A

A B (b) C D

Arrangement of optical system in polarimeter:

The instrument used for measuring the optical rotation is known as polarimeter. The Lippich
polarimeter is generally adopted nowadays, the arrangement of various optical parts is
diagrammatically shown in the figure.

L is the source of monochromatic light. The light from L passage through A, which makes the
ray of light parallel.

It then passes through the polarizing prism B, through the observation tube O and then
through the analyzer D, E, F is the telescope from which the field of view is observed. C is a
small Nicol prism which covers half of the opening at the end of polarizer tube.

On passing through this prism the light is altered a phase by half a wave length, but still
remain plane polarized. Thus two beams of polarized light are obtained. If the polarizer is
rotated so that the plane of polarization forms an angle  with the axis of the Nicol prism, the
plane of polarization will also be inclined to an angle 20. This is known as half shadow angle.

When the analyzer is rotated, a position is found at which one beam will be completely, the
other only partially, extinguished. Therefore the one half of the field of view will appear dark
and other half will still remain in light. On rotating the analyzer further through the angle 20,
the position B is first is no longer so. At this position, the half of the field which was formerly
bright will now be dark and that formerly dark will now be light. At the intermediate position
C, the field of view will be of uniform brightness. The analyzer must be set in this position
while working. In certain polarimeters the triple field D is observed.

The scale is generally graduated directly into quarters of degrees and with the help of
verniers and magnifying glass, the angle may be read to 0.01. By diminishing the angle 
the sensitiveness of the instrument is increased.

For most of the laboratory work, monochromatic yellow light of incandescent sodium vapor is
used as the source of light. The source of light should be placed at the focal distance from
the end polarimeter at about 20cm. The observation tube is generally 10cm long.

20
Determination of zero points

Clean the polarimeter tube with chromic acid and finally with water. Place the light source at
the suitable distance. Focus the telescope eye piece and rotate the analyzer until the
position C is obtained. Note down each side of the scale. It gives the right and left zero
points. These zero readings should be taken at the beginning of the experiment and must be
subtracted from the corresponding readings of solution of the optically active substance.

Determination of angle of rotation

Prepare a dilute solution (about10%) of the substance. Rinse the observation tube with the
solution and fill it with the solution with the help of glass plate. The tube should be
completely free from any air bubble. Determine the position C by rotating the analyzer as
discussed in the introduction.

The difference between the reading on the right and the zero on the right gives the angle of
rotation ().

Since the circular scale may not be exactly centered, one should take the average of
readings on the right and the readings on the left side.

Find out the specific rotation at the temperature of the experiment using equation. The
difference between this reading and zero reading is the angle of rotation . Similarly
determine the angle of rotation of 15%, 20%, 30% solutions of cane sugar.

Observed Data

Length of polarimeter tube = cm

Temperature of experiment = C

Zero correction = Degrees

S. No. Conc. of Observed Corrected Specific

solution rotation rotation () rotation []t

1 10%
2 15%
3 20%
4 30%
Mean

Apparatus & Chemicals required:

Polarimeter, sodium vapour lamp, sugar solution, measuring flasks etc

Procedure:
Set up the sodium vapor lamp at a distance equal to the focal length of the lens (about 10cm)
so as to get a uniform illumination of the polarizer.

21
Clean the observation tube with chromic acid and finally with distilled water. Fill it with
distilled water without any air bubble and place it in the polarimeter. Now by the help of
rotating disc obtain the uniform illumination with a barely visible central line on right and left
of the scale. This is zero reading, which must be subtracted from the final reading of the
solution.

Now prepare 10% of the cane sugar solution in water. Rinse the polarimeter tube with the
sugar solution and then fill it with the same. Now rotate the analyzer to get the position of
uniform illumination again. If the field is dark on the right, the substance is dextro rotatory
otherwise leavo rotatory on the left. Note this reading.

Precautions

a) Clean the polarimeter tube and glass plate every time and dry it.

b) The solution should be transparent; if necessary it should be filtered.

c) There should be no air bubble while filling the polarimeter tube.

d) When the experiment is over, the tube should be well washed and the caps are
un-screwed.

Calculation and results:

Specific rotation of different concentrations of food sugar has been estimated following
polarimetric method.

The value of []t  may be calculated from

t  100
l.c.

Where l is the length of polarimeter tube in decimeters and c is the concentration of the sugar
in 100ml of solution.

Kinetics of hydrolysis:
A 20% of sucrose is prepared and a drop of thymol is added to preserve it. Exactly 20 ml of
sugar solution and exactly 20 ml of 1N HCl acid solution are mixed after noting the initial time
of mixing. The reaction mixture is poured in a polarimeter tube. The angle of rotation is noted
at regular intervals, say, 10 minutes. The reaction is allowed to go to completion and the final
angle of rotation is noted.

Let the initial angle of rotation at the start of the reaction is α0 , the rotation at any instant

is αt ,and the infinite reading be αα Thus we have initial concentration of sugar,

α0=(α0-α∞); at time t, (a-x) = (αt- α∞)

Knowing the various values at different time intervals, k1’ is calculated which should be
constant.

Observations and Calculations

22
 Time Angle of 2.303
S. No (αt - αα)
( sec ) rotation (α) k1’ = t log (α0 - α∞)/(αt - α∞) sec-1
1. 0 α0
2. α1
3. α2
4. α3
5. α4
6. α5
7. α6
8. ∞ α∞
Mean k1’ = -----------------sec-1

Graphical Method

Log (αt - α∞) Time (sec)

Log (a-x)

Time (secs )

Inversion of sucrose

K1’ = slope x 2.303

𝑑𝑦
𝐾′1 = × 2.303
𝑑𝑥

= ____________ sec-1

Ratio of rate constants = k2/k1 which is equivalent to the ratio of the acids.

Results:
1. The specific rotation of given sugar sample =

𝐾
2. The ratio of rate constants of inversion of sucrose (𝐾1 ) =
2

3. The ratio of strength of given acid =

23
Experiment 5: Building Voltaic Cells or a stable
Battery from Waste Metal
Aim:
Many objects and tools made of metals which are discarded as waste. The metals are to be
utilized to build a voltaic cell or battery using various household objects with the help
chemistry laboratory.

Principle:
A galvanic cell uses the energy released during a spontaneous redox reaction (ΔG < 0) to
generate electricity. Overall representation of Galvanic cell is presented as M1 | M1 n+ (C1) ║
M2m+ (C2) | M2 where M1 metal is oxidized and M2m+ is getting reduced. During the process
the produced electricity can be presented as
𝐸𝑐𝑒𝑙𝑙 = 𝐸𝑀2/𝑀2𝑚+ − 𝐸𝑀1/𝑀1𝑛+
𝑜 𝑜 0.059 0.059
𝐸𝑐𝑒𝑙𝑙 = 𝐸𝑀2/𝑀2 𝑚+ − 𝐸𝑀1⁄𝑀1𝑛+ − 𝑙𝑜𝑔10 [𝑀1𝑛+ ] + 𝑙𝑜𝑔10 [𝑀2𝑚+ ]……….. (1)
𝑛 𝑚

The simplified equation is meant for the approximated EMF of the constructed battery. The
equation may not always stand for M2 metal considering the oxidation of Fe in presence of
H2O and O2. For that reaction the equation (1) can be written as
0.059 0.059
𝐸𝑐𝑒𝑙𝑙 = 𝐸𝑂𝑜2−⁄𝑂 − 𝐸𝐹𝑒/𝐹𝑒
𝑜
2+ − 𝑙𝑜𝑔10 [𝐹𝑒 2+ ] + 𝑙𝑜𝑔10 [𝑂]……….. (2)
2 2

From the electrochemical series where, 𝐸𝑂𝑜2−⁄𝑂 = −0.83 & 𝐸𝐹𝑒/𝐹𝑒

𝑜 𝑜
2+ = −0.45, 𝐸𝑐𝑒𝑙𝑙 = −0.38𝑉.

From the relationship ∆𝐺 = −𝑛𝐹𝐸𝑐𝑒𝑙𝑙 the reaction is non spontaneous. We can also find that
𝐸𝑂𝑜2−⁄𝑂 = 1.83𝑉 when the pH of the electrolytic medium becomes less than 7 due to protic
𝑜
acid. Hence for acidic medium 𝐸𝑐𝑒𝑙𝑙 = 2.25𝑉 make the redox reaction spontaneous to make
a galvanic cell to produce electricity.
Requirements:
Reagents and solutions: Metal from waste object, Pencil lead for Carbon electrode, NaCl,
Acetic Acid etc.
Apparatus: Potentiometer, Standard Calomel Electrode (SCE), 100 mL beakers, salt bridge.
5mm multicolor LEDs, 70 Points Mini Breadboard, Solderless Mini Breadboard for
Prototyping, solid core wire for breadboard, Alligator (Crocodile) Clip and Vernier scale

Procedures:
The metallic object is washed well with distilled water. 50 mL of the electrolyte solution is
taken in a beaker and its corresponding metal electrode is introduced. As day-today metallic
objects are made of one among the common metals like Fe, Al and Zn, students must be
intuitive to choose electrolyte solution accordingly. 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 from where
an electrode potential of waste metal [EM/M+] is estimated.

24
Table 1: The estimation of waste metals’ electrode potentials [[EM/M+]

Sl Waste Ecell (V) (EMF of Chosen Electrolyte solvent & EM/M+ = Ecell +
No. Metals the cell) Concentration (N) Ecalomel
1
2
3

From the estimated electrode potentials, make a table for the of cathode and anode
combination and complete the table 2.

Table 2: The choice of metal electrode combination to build series of cell or battery

Sl Waste Chosen Electrolyte solvent & Ecell (V) from Experimental Ecell
Combinations
No. Metals Concentration (N) table 1 (V)
1 M1 M1 & M2
2 M2 M2 & M3
3 M3 M3 & M1

The pair of metals or electrodes after estimating as per the

After estimating as per Table 1, the pair of metals or electrodes can be tested by building the
galvanic cells, as represented in Figure 1. The highest obtained Ecell can be chosen to
connect in series for glowing a 3.5V LED. Once the battery is built, the output voltage is
checked with a time interval of 4 minutes for 20 minutes to realize its stability, and the
respective data will be tabulated in Table 4

Figure 1: A Representative of constructed cell

25
 Table 3: Result of adequate cells in series to generate 3.5 V and glowing LED

No. of estimated cells as per No. of required cells to glow Measured EMF of constructed battery
Table 2 LED (V)

Table 4: Result of Voltage stability of built battery

Sl No. Time (min) Measured EMF of constructed battery (V)

1 0
2 4
3 8
4 12
5 16
6. 20

26
Experiment 6: Building Dye Sensitized Solar Cells as
Renewable Resource from Natural Dye
Aim:
To harvest solar energy by using TiO2 powder and readily available natural dye from
pomegranate, beet root, carrot etc. along with ITO coated substrate.

Principle:
Many fruits and flowers are rich in natural dyes with different colours indicating different
wavelength of absorption. On varrious occasion it has been found that parts of the fruits
containing dye are discarded and the flowers after their decorative or ritual purposes become
waste products. In Dye Sensitized Solar Cell (DSSC) such natural dyes can be the worthy to
use as absorber layer in combination with TiO2 which is commonly utilized for electron
acceptor or photo anode of the cell. In DSSCs light is absorbed by a monolayer of a dye
located at the junction between an electron and hole transporting phases where the electron
transport material is a wide band gap semiconductor material typically TiO2 (anatase), while
the hole transport system is a redox shuttle. On light-irradiation the dye injects an electron
and a hole into the n- and p-type materials, respectively, generating free charge carriers,
which then travel through the electron acceptor to be collected as current and flow to the
external circuits. During the last two decades research efforts have been made on the
development of new sensitizers, photoanode engineering, redox shuttle and catalyst to
fabricate efficient devices. The DSSC consists of a mechanical support coated with a
transparent conductor, on which TiO2 powders are deposited to form a mesoporous network
on which a sensitizer is anchored, and in combination with graphite cathode separated by a
thin layer of redox shuttle.

Fig.1: Operating principles and energy level diagram of a dye-sensitized solar cell

27
 Requirements:
Reagents and solutions: ITO coated PET or Glass, natural dye, TiO2 powder, conducting
charcoal. KI, I2 solution, EtOH, and polymer adhesive, transparent PET sheet or glass slide.

Apparatus: Multimeter, Hotplate. 100 mL beakers, glass rod, 5 mm multicolor LEDs, 70

Points Mini Breadboard, Solderless Mini Breadboard for Prototyping, solid core wire for
breadboard, and Alligator (Crocodile) Clip

Procedure:

Commercially available ITO-coated PET sheets or ITO-coated glass substrates have to be

coated with TiO2 ethanol slurry mildly acidified with dilute HNO3. Then, the coated film will
be dried on a hot plate adjusted to 60 °C for 5 minutes, and after drying, it will be soaked
with natural dye (pomegranate juice or ethanol/water extract of red hibiscus flower, etc.).
The soaked TiO2 film on ITO will be dried or backed at 70 °C for 5 minutes. The conducting
graphite powder or Carbon slurry of polyols or organic gums will be coated on a PET sheet or
glass slide. After drying, the cathode made of carbon will be soaked in KI+I2 solution. The
redox solution-soaked conducting carbon film and dye-soaked TiO2 films are sandwiched so
that the cathode and anode face each other, leaving the edges to connect the wire for
measuring a voltage drop across the electrodes. The transparent side of the ITO-coated film
is illuminated under sunlight, and the voltage drop will be measured through a multimeter.
The overlapped area of the cathode and anode is to be measured in square cm, and based
on the voltage drop, how many cells are to be connected in series to reach 3.5 V is to be
calculated.

Results:

Name of Measured Voltage Minimum number of required

Number of Overlapped
the Dye, Drop by the cell made number of cells for generating
cells made Area (cm2)
used (V) 3.5 V

28
Experiment 7: Fabrication and Evaluation of a
Supercapacitor from the Biowaste Derived from
Sweet Lime Peels
Aim:
To synthesize activated carbon (AC) from sweet lime-peel bio-waste and determine the
diffusive controlled and capacitive current currents of the fabricated AC-based
supercapacitor.

Materials and reagents:

Sweet lime peels, polyvinyldene fluoride (PVDF), N-methyl pyrolldine (NMP), nickel foam,
nafion, Cu wires as connector, Hg/HgO Reference Electrode, Platinum foil.

Principle:
Batteries and capacitors do a similar job—storing electricity—but in completely different
ways. A capacitor is a device used to store electrical charge and electrical energy. Capacitors
use static electricity (electrostatics) rather than chemical substances to store energy.
Inside a capacitor, there are two conducting metal plates with an insulating material called a
dielectric in between them - it's a dielectric sandwich. Positive and negative electrical
charges build up on the plates and the separation between them, which prevents them
coming into contact, is what stores the energy. The dielectric allows a capacitor of a certain
size to store more charge at the same voltage, so it makes the capacitor more efficient as a
charge-storing device.
I ∆t
Specific capacitance Cs = m ∆V

where I is the current applied, m is the mass of the active material, Δt is the discharge time
and ΔV potential between the two parallel plates of the capacitor.

A supercapacitor differs from an ordinary capacitor in two important ways: Its plates
effectively have a much bigger area and the distance between them is much smaller,
because the separator between them works in a different way to a conventional dielectric.
Although the words "supercapacitor” and "ultracapacitor” are often used interchangeably,
there is a difference: they are usually built from different materials and structured in slightly
different ways, so they store different amounts of energy.

Like an ordinary capacitor, a supercapacitor has two plates that are separated.

The plates are made from metal coated with a porous substance such as powdery,
activated charcoal, which effectively gives them a bigger area for storing much more charge.
Unlike capacitor, in a supercapacitor, there is no dielectric as such. Instead, both plates are
soaked in an electrolyte and separated by a very thin insulator (which might be made of
carbon, paper, or plastic)

29
Procedure:
Synthesis of activated carbon:
Cut the fresh sweet lime peels into small pieces and wash with tap water several time
followed by de-ionized water and then dry in a hot-air oven at 80oC overnight. It was then
crushed using an electrical grinder to obtain a fine powder. For activation, 10 grams of sweet
lime peel powders were immersed in 1M KOH for 1 hour. Later, the soaked sweet lime peel
powder was separated from the mixture, and the KOH solution was removed. The wet sweet
lime peel powder was rinsed with de-ionized water and dried for 10 hours at 80°C. For
carbonization, the KOH-treated sweet lime peel powder was maintained in a muffle furnace
at 500°C for one hour. The carbonized material was then collected and washed with 1 M HCl
to eliminate any organic contaminants. Later, it was washed many times with de-ionized
water, ethanol, and dried in a vacuum oven at 70°C for 12 hours to obtain activated carbon
(AC).

Fabrication of Capacitor:
The working electrodes for electrochemical evaluation will be prepared by mixing 90 wt. % of
activated carbon black synthesised and 10 wt. % polyvinylidene difluoride (PVDF) binder in
an agate mortar. The mixture will be made as semi-solid slurry by the addition of a few drops
of 1-methyl-2-pyrrolidinone (NMP). The paste was coated on the Ni foam (1 cm x 1 cm) as a
current collector and dried at 90 °C in an oven for 8 hours to ensure complete evaporation of
the NMP.

30
Electrochemical studies were carried out on a Palmsense potentiostat/galvanostat (model:
Emstat HR) using a conventional three-electrode system. To understand the capacitance
behavior of as-synthesized activated carbon, the CV measurement should be performed in
3.0 M KOH electrolyte. Applied potential was scanned in the 0.0 to 0.6 V range vs. Hg/HgO
reference electrode with 50 mVs-1 scan rate will be applied. The scan rate will be also varied
from 1, 4, 8, 12, 16 and 20 mV/s in order to verify the diffusion controlled process of the
hydroxide ions. Galvanostatic charge discharge (GCD) curves will also be recorded at
different current densities applied and measured the variation in specific capacitance with
applied current density.

Here, we apply the Dunn method to calculate the capacitive current and diffusion controlled
current of the CVs recorded.
According to the 𝑖
equation, 𝑣 1/2
= k1ν1/2 +k2

Where, ‘i’ is the current, ν is the scan rate, k1 and k2 constants corresponding capacitive
current and diffusion current.

Steps for Recording CVs:

1. Fill the electrochemical cell with 10 ml of 3.0 M KOH

2. Connect the three electrode system, with AC-coated Ni-foam to working electrode, Pt
foil as counter electrode and Hg/HgO reference electrode probes.
3. Record the cyclic voltammograms at different scan rates ca. 1, 4, 8, 12, 16 and 20
mV/s
4. Plot and fit the graphs between i/ν1/2 and v1/2 for different scan rates. From the y-
intercept of the plot, and determine the diffusion controlled current.

Results:

Weight of activated carbon material synthesized = ------- gm

Percentage of diffusion controlled current of the capacitor =------- %

Percentage of capacitive controlled current of the capacitor = ------ %

31