Challenging experiments lab manual

S.No Experiment Page

1 Comprehensive Water Analysis – Environmental analyses 2
2 Determination of the Calorific Value of any energy source (fuels or food 8
products) by Bomb Calorimeter – Energy (Chemical and bio-) source
evaluation
3 Synthesis and characterisation of hydroxyapatite nanoparticles from 11
eggshell biowaste by chemical precipitation and study its application for
Pb (II) removal from aqueous wastewater. – Waste to wealth
4 Determination of specific rotation by polarimetry- Kinetics of inversion of 15
sucrose in normal sugar and refined sugar samples – Food quality
Experiment 1: Comprehensive Water Analysis
Aim:
Collect a locally available water sample (suspected for contamination) and analyze its
hardness, chloride content, Total Dissolved Solids (TDS), pH, and sulfate content in it. After
analyzing compare the water quality as specified by Central Pollution Control Board, Ministry
of Environment, Forest and Climate Change, Government of India
(https://cpcb.nic.in/wqstandards/)

Principles:
a) Hardness Estimation: 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. (reference present
Manual)

b) Chloride Estimation: the methods considered for chloride analysis in this study are
based on the well-known reaction between chloride ions and AgNO3, when a white
precipitate of AgCl is formed, according with the following equation:

Cl- + AgNO3 = AgCl+ NO3- (2)

The obtained precipitate has a low solubility in water. In addition, because only halide and
sulfide ions precipitate in presence of AgNO 3, such methods have a good selectivity in
quantitative analysis of chloride ions. The representative titration graph is given below.
c) Total Dissolved Solids (TDS) estimation: 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)=k e × EC (μS/cm) (3)

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. (Reference: https://pure.psu.edu/en/publications/relationship-between-total-
dissolved-solids-and-electrical-conduc or https://doi.org/10.2166/wst.2018.092)

d) pH Estimation: 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. (Reference: https://nitsri.ac.in/Department/Civil
%20Engineering/CWE-301_WATER_QUALITY_AND_ENVIRONMENT_pH_Meters.pdf)

e) Sulfate Estimation: Sulfate ion is converted to a barium sulfate suspension under

controlled conditions. The resulting turbidity is determined by a nephelometer, filter
photometer, or spectrophotometer and compared with a curve prepared from standard
sulfate solution. (Reference:
https://www.epa.gov/sites/default/files/2015-12/documents/9038.pdf)
−¿¿
2−¿=BaS O4 ↓+2 Cl ¿
BaCl 2+ SO4 (4)

Requirements:
a) Hardness estimation: 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.

b) Chloride Estimation: 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

c) Total Dissolved Solids (TDS) estimation: Reagents and solutions: None. Apparatus:
Commercial TDS meter, Pt electrode, and a digital conductometer.

d) pH Estimation: Reagents and solutions: pH buffer solution for calibrating pH, and pH
paper. Apparatus: Digital pH Meter along with electrodes. Commercial TDS meter, Pt
electrode, and a digital conductometer.

e) Sulfate Estimation: Reagents and solutions: conc. HCl, Isopropanol, NaCl, Propanol,
glycerol, Na2SO4, and BaCl2.

Apparatus: Magnetic stirrer, Nephelometer, Stopwatch, Measuring spoon, 250 mL conical

flask, and 1L volumetric flask.

Procedures:

a) Hardness estimation:
Pipette out 20 mL of the given sample of 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. No. Volume of sample Burette reading (ml) Volume of EDTA

hard water (ml) Initial Final (V2, ml)
1
2
3
Concordant titre value (ml)

Calculations:

If EDTA is standardized as 1 mL of EDTA requires = w mg CaCO3 for complexation

From the above titration 20 mL of sample water consumes = V2 mL of EDTA = V2 x w mg of

CaCO3 eq.

\1000 mL of hard water sample consumes = V2 x w × 1000/20 = V2 x w × 50 ppm

\Hardness of the water sample = “V2 x w × 50” ppm

b) Chloride Estimation:

Make up the unknown water solution given in a 100 mL standard flask up to the mark using
distilled water. 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 fill the burette with ~0.01 N AgNO 3 solution (from
Bottle B). Record the conductivity of the water sample without adding any AgNO 3 solution
from the burette (0th reading). Add 1 mL AgNO 3 solution of known concentration into the
beaker with continuous stirring by magnetic stirrer. Continue the addition of AgNO3 solution (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 AgCl, and then starts increasing on continuing the
addition of AgNO3. A graph is now drawn by plotting conductance vs volume of AgNO 3 added.
Intersection point from the plot gives the volume of AgNO3 required for precipitating the
chloride present in the water sample.

Estimation of unknown chloride:

(N x V) of water sample = (N x V) of AgNO3 solution

N of irrigation water sample = N of AgNO3 x Volume measured from Plot (Ve)

20 mL= N of water sample

Amount of chloride present in 1L = Normality of irrigation water sample x Eq. wt. of Cl -

(35.453)
Amount of chloride present in given sample solution =
Strength of water sample x 35.453 x 100
1000

= grams in 100 mL

Result: Amount of chloride in given water sample =

Stre ngth of water sample x 35.453 x 1000 ppm

c) Total Dissolved Solids (TDS) estimation: Recorded the conductivity of the water sample
without adding any AgNO3 solution from the burette (0th reading). The recorded conductivity
when multiplied by 0.7 (ke) as per the equation

TDS (mg /L)=k e × EC (μS/cm)

TDS is estimated in terms of ppm.

d) pH Estimation: The pH electrode to be dipped in a beaker of sample water and pH will be

displayed and to be recorded. Avoid any kind of contamination and use a clean and dry
beaker before pouring the sample water.

e) Sulfate Estimation:

i.) Preparation of conditioning reagent: Slowly add 30 mL concentrated HCl to 300 mL of DI

water, 100 mL 95% ethanol or isopropanol, and 75 g NaCl in solution in a container. Add 50
mL glycerol and mix.

ii.) Formation of barium sulfate turbidity: Place a 100-mL sample, or a suitable portion diluted
to 100 mL, into a 250-mL Erlenmeyer flask (conical flask). Add exactly 5.0 mL conditioning
reagent and mix in the stirring apparatus. While the solution is being stirred, add a measured
spoonful of BaCl2 crystals and begin timing immediately using stop watch. Stir exactly 1.0
min at constant speed.

iii.) Measurement of barium sulfate turbidity: Immediately after the stirring period has ended,
pour solution into absorbance cell. Measure turbidity at 30-sec intervals for 4 min. Record the
maximum reading obtained in the 4-min period.

iv.) Preparation of calibration curve: Dissolve 147.9 mg anhydrous Na 2SO4 in DI water in a 1-

liter volumetric flask and dilute to the mark for standard sulfate solution. Space standards at
5-mg/L increments in the 0-40 mg/L sulfate range. Prepare a standard curve by plotting
turbidity (NTU units) vs the sulfate range. From the calibration plot, measure the sulfate
concentration in the given unknown sample.
 Report of Water Sample Analysed
Sample Collected from
Date of Collection

Recommended Values as
Recorded Deviation from the
Parameters per BIS (Bureau of Indian
Values standard (%)
Standards)
Hardness (ppm)
Chloride content (ppm)
TDS (ppm)
Sulfate content (ppm)
pH
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
 ( W 1+ W 2 ) ×(t 2−t 1)× 4.184
GCV = kJ /kg …….. (1)
m

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 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 ×m B
W 2= −W 1 g
(t 2 −t 1 )× 4.184

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)

( W 1+ W 2 ) ×(t 4 −t 3)×4.184
GCV fuel = kJ /kg
m fuel

=……………………………. kJ/kg

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)

( W 1+W 2 ) ×(t 5−t 6 )× 4.184

GCV food = kJ /kg
m food

=……………………………. kJ/kg
Experiment 3: Removal of heavy metals from waste
water using nanoparticles prepared from bio-waste
Aim:

To synthesize and characterize hydroxyapatite nanoparticles from eggshell bio-waste by

chemical precipitation and study its application for Pb (II) removal from aqueous 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. HAP is a bioactive ceramic that has gained wide attention in recent years
owing to its potential applications in several fields, such as bio-ceramics, medicine, the
pharmaceutical industry, and environmental pollution control [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 amounts 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 [3]. 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, 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) [4].

Requirements:
 Cleaned the eggshell
 Concentrated HNO3
 NaOH
 Ca(NO3)2
 (NH4)2HPO4

Procedures:
Natural bio-waste precursors for HAP synthesis- HAP nanoparticle synthesis from eggshell
bio-waste:
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 HNO 3 slowly. Stir the mixture
continuously until the eggshell powder dissolves completely in nitric acid. To this, add 15 ml
of deionized water followed by 20 ml of 4N NaOH solution to get yellow coloured colloidal
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 0.1 M of (NH 4)2HPO4 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:

Characterization:
The powder XRD pattern of the collected powder sample was recorded on a Rigaku
Smartlab 3 KW X-ray diffractometer with Cu Kα radiation source (λ=1.54060 Å)) in the scan
range 10-80 (2). The standard XRD pattern of HAP is given in Fig. 1 for reference.
 Fig.1. XRD pattern of HAP nanoparticles

Analysis:
The powder XRD peak positions for the prepared HAP nanoparticles by chemical
precipitation method are identified based on standard JCPDS file #__09-0432.

Particle size calculation using Scherrer’s equation:

Scherrer equation
Grain size = k* / cos(θ)*(FWHM)
k = 0.9,  = 1.0506 Å, and “” and FWHM will be obtained from powder XRD data.

Removal of Pb2+ ions from wastewater:

The lead (Pb2+) ions from wastewater can be removed by batch adsorption study using HAP.
First, 0.1 g of HAP as an adsorbent is added to 100 mL of wastewater containing Pb 2+ ions
and shaken (180 rpm) for 1 h at room temperature. Afterwards, the HAP is filtered using
whatmann filter paper and the residual lead content remaining in the filtrate was assayed by
using a UV visible spectrophotometer in the presence of dithiazone solution (0.001% in
chloroform and ethanol as a stabilizer) at a wavelength of 530 nm. Finally, the percentage of
adsorption Pb (II) ions on HAP nanoparticle are calculated based on initial and final
concentrations of Pb2+ ion in the wastewater and using the following equation.
(C o−C e )
% Removal = × 100
C0

Results:
(i) The powder XRD peak positions for the prepared by HAP nanoparticles are
____________________________________________.
(ii) Particle size ____________________________________________.
(iii) The percentage removal of Pb(II) ions from aqueous medium is
-------------------------------------.
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 ‘[]’.

Where l = length of column of liquid in decimeter and

d = density

Taking into account the factors affecting angle of rotation.

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 C B A
o

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 now a days, 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.
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.
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

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-αα); at time t, (a-x) = (αt- αα)

Knowing the various values at different time intervals, k 1’ is calculated which should be
constant.

Observations and Calculations

 Time Angle of 2. 303
S. No (αt - αα)
( sec ) rotation (α) k1’ = t log (α0 - αα)/(αt - αα) sec-1
1. 0 α0
2. α1
3. α2
4. α3
5. α4
6. α5
7. α6
8. α αα
Mean k1’ = --------------- sec-1

Graphical Method

Log (αt - αα) Time (sec)

Log (a-x)

Time (secs )

Inversion of sucrose

K1’ = slope x 2.303

dy
K1’ = dx x 2.303

= ___________ 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 =
 ( )
k1
k2
2. The ratio of rate constants of inversion of sucrose =

3. The ratio of strength of given acid =