Minor Project Report

On
Analytical Chemistry

Karan Gupta
Roll No: 2110404010306
B.Sc. Chemistry Semester-VI

Submitted to;
Dr.Niranjani Chaurasiya
Department of Chemistry
Shri Jai Narayan P.G. College(KKC)
Station Rd, Khatkeyana, Lucknow,
Uttar Pradesh
 ACKNOWLEDGMENT

I would like to extend my heartfelt appreciation to the Principal, the Head of the Chemistry
Department, and my project supervisor for their continuous support and encouragement
during every phase of my minor project in chemistry. I would like to convey my sincere
gratitude to my Principal Prof. VINOD CHANDRA and H.O.D SHRI AJAI KUMAR
MISHRA ,Department of Chemistry J.N.M.P.G College, for providing me with the essential
resources and infrastructure finishing my project.

I would like to express my gratitude to my research project supervisors, Prof. NIRANJANI

CHAURASIYA, Department of Chemistry, (KKC), for their invaluable guidance, constant
support and mentorship throughout the duration of this project,, and helping me navigate
challenges along the way. Their expertise and mentorship have not only enhanced the quality
of my project but have also broadened my understanding of the subject.

I feel privileged to have had the opportunity to work under their guidance and am thankful for
the knowledge and skills I have gained through their mentorship. Their unwavering support
has been instrumental in shaping my research, and I am truly grateful for their contributions
to my academic development.

Once again, I express my sincere gratitude to the Principal, the Head of the Chemistry
Department, and my project supervisor for their exceptional support and guidance throughout
my minor project in chemistry.

[KARAN GUPTA ]
 INDEX

Contents
 ACKNOWLEDGMENT .................................................................................................................. 0
 INDEX.............................................................................................................................................. 0
 THIN LAYER CHROMATOGRAPHY (TLC)................................................................................ 1
INTRODUCTION ..................................................................................................................... 1
DISCUSSION .......................................................................................................................... 17
REFERENCE........................................................................................................................... 18
 HARDNESS OF WATER.............................................................................................................. 19
INTRODUCTION ................................................................................................................... 19
DISCUSSION .......................................................................................................................... 30
REFERENCE........................................................................................................................... 32
 pH OF BUFFER SOLUTIONS ...................................................................................................... 33
INTRODUCTION ................................................................................................................... 33
pH............................................................................................................................................. 33
BUFFER SOLUTION ............................................................................................................. 36
DISCUSSION .......................................................................................................................... 43
REFERENCE........................................................................................................................... 45
 POLARIMETER ............................................................................................................................ 46
INTRODUCTION ................................................................................................................... 46
DISCUSSION .......................................................................................................................... 58
REFERENCE........................................................................................................................... 60
 THIN LAYER CHROMATOGRAPHY (TLC)
INTRODUCTION

TLC is one of the simplest, fastest, easiest and least expensive of several chromatographic
techniques used in qualitative and quantitative analysis to separate organic compounds and to
test the purity of compounds[1]

Thin-layer chromatography (TLC), is a solid-liquid form of chromatography where the

stationary phase is normally a polar adsorbent and the mobile phase can be a single solvent or
combination of solvents.

In thin-layer chromatography, the stationary phase is a polar absorbent, usually finely ground
alumina or silica particles. The adsorbent is coated on a glass slide or aluminium sheet or plastic
sheet creating a thin layer of the particular stationary phase. Almost all mixtures of solvents
can be used as the mobile phase. By manipulating the mobile phase, organic compounds can
be separated.

In TLC, molecules are continuously moving back and forth between the free and adsorbed
states with millions of molecules adsorbing and millions of other molecules desorbing each
second.

The equilibrium between the free and adsorbed states depends on three factors:

• The polarity and size of the molecule

• The polarity of the stationary phase
• The polarity of the solvent

The polarity of the molecules is determined by their chemical structures. By selecting different
stationary and mobile phases, we can change the equilibrium between the free and adsorbed
states. Different molecules behaves partition differently between the free and adsorbed state
that is the equilibrium between these two states is not the same.

The stationary phase used in TLC is Silica gel (SiO2 .xH₂O)n. The covalent network of these
adsorbents creates more polar nature.

TLC is a form of liquid chromatography consisting of:

1
  Analysis is performed on a flat surface under atmospheric pressure and room
temperature

Chromatography means color writing. It is a separation technique working on the principal of

differential distribution of components of a mixture between the mobile phase and the
stationary phase. Individual components of the synthesized compound move through the
packed bed of the stationary phase at different rates. The least hindered component elutes first
while highly hindered component elutes at last. All chromatographic techniques work on the
principle of adsorption or partition.

When the stationary phase is a Solid adsorbent like silica or alumina and mobile phase is a
liquid the process is called adsorption chromatography. [1]

These are of two types:

I. Thin layer chromatography

II. Paper chromatography

Comparison between Thin Layer Chromatography and Paper Chromatography

Thin Layer Chromatography Paper Chromatography

Principal of separation is adsorption Principal of separation is partition

More quantity of sample is required Minimum quantity of sample is required

Consuming less time (15-45 min.) Consuming less time (1-3 h)

Plates may be heated in oven for long Paper cannot be heated for long time
time
Very sharp separation Less sharp separation

Physical strength of the plate is high
(Ascending/Descending)
Plates are prepared as thin coat with
adsorbents
Corrosive reagent can be used
Compound can be detected under UV
Physical strength of the paper is low
(Descending/ Small paper only
Ascending)
Paper is used as such since coating
can’t be done on paper
Corrosive agent can’t be used since it
destroys it
Can’t be detected under UV

Sensitivity of detection is high Less sensitivity of detection

2
Principle
Process involves the spreading of the thin layer of adsorbent (stationary phase) on a suitable
support which may be either on glass plate, aluminium foil or a plastic sheet. But commonly
the glass plates are used in the laboratory.[9]

Sample is applied and when the solvent has evaporated off, the plate is placed vertically in a
TLC chamber containing the mobile phase. When the mobile phase reaches the necessary
distance, the plate is taken out, dried in air and developed for detection.

Position of each component on the plate is expressed by its Rf value:

Rf = Distance Travelled By the Component

Distance Travelled By the Solvent

Procedure
1. Choice Of Stationary phase (Adsorbent) – Most commonly used stationary phase in TLC
is silica gel and Alumina. All the adsorbents contain binder which is usually Plaster of Paris
which holds the adsorbent firmly on the plate

2. Preparation of thin layers on glass plates - The solid stationary phase on the glass plate
can be prepared by any one method-

(a) Pouring

(b) Dipping

(c) Spraying

(d) Spreading

3
Pouring

A measured amount of slurry is put on a given size of plate, which is kept on a level surface.
The plate is then tipped back and forth to spread the slurry uniformly over the surface.

Dipping

In the dipping method, the TLC plate is vertically immersed in the dramatization reagent for
several seconds.

Spraying

Slurry is diluted further for the operation of sprayer. But this technique is not used now a days
it is difficult to get uniform layer

Spreading

This is the most common for obtaining thin uniform layer of adsorbent on the glass plate. In
this technique, several glass plates are cleaned and dried. A slurry (in water) of silica gel is
made and poured on the glass plate and then spread over with a glass rod. The coated glass
plates are air dried and then processed to remove surplus adsorbent from the edges.

Methods of preparation:-

1. TLC plates used are purchased as 5 cm x 20 cm sheets. Each large sheet is cut
horizontally into plates which are 5 cm tall by various widths

2. Handle the plates carefully so that you do not disturb the coating of adsorbent or get
them dirty. Measure 0.5 cm from the bottom of the plate.

3. Using a pencil draw a line across the plate at the 0.5 cm mark. This is the origin: the

4
4. Under the line mark highly the samples you will spot on the plate or mark numbers for
time points leave enough space between the sample so that they do not run together about 4
samples on a 5 cm widely plate is advised.

3. Loading of sample

Prepare the sample solution by dissolving around 2 mg of compound in few drops of suitable
solvent (i.e., ethyl acetate).

Now dip the capillary tube into the sample solution and then bring the capillary near the TLC
plate and lightly touch it on the silica gel. Now remove the capillary tube and allows the solvent
to evaporate. This should be done in the middle of the TLC plate and about 0.5 – 1 cm from
the bottom of the plate. [9]

Spot the TLC plate

• Prepare 1% solution of drug dissolving in volatile solvents like hexanes, ethyl acetate,
or methylene chloride.
• Dip the microcap or micro capillary into the solution and then gently touch the end of
it onto the proper location on the TLC plate.
• Don't allow the spot to become too large - if necessary, you can touch it to the plate, lift
it off and blow on the spot. If you repeat these steps, the wet area on the plate will stay
small.
• This example plate has been spotted with three different quantities of the same solution
and is ready to develop

5
4. Choice of Mobile Phase

a) The sample should be at least moderately soluble in the mobile phase

b) The sample and the mobile phase (solvent) should not react with each other

c) The solvent used for running the TLC should make the sample move halfway up the plate

d) The mobile phase can be a single solvent or a combination of solvents

5. Detection

Detection of the separated compound is the final step of TLC.

a) Physical methods: The developed plate is observed in daylight and then under UV

b) Chemical Methods: A Suitable reagent giving color spot is sprayed on the TLC plate.
Spraying can be done with a sprayer. Some Universal spraying agents are 50% aqueous
solution of H2SO4, H2SO4 mixed electron suitable aldehyde, sodium or potassium
dichromate etc.

Iodine is also used for the detection process. Crystals of iodine are placed in the iodine chamber
and then the developed TLC plate is placed in this iodine chamber for few minutes.
Vapors of iodine form charge transfer complexes with many organic compounds.

Two Step Synthesis:-

6
Step 1 Synthesis:-

Synthesis of Phthalimide from phthalic anhydride was carried out as per the procedure and
the TLC was carried out for both the starting compound (Phthalic anhydride) and final
product (Phthalimide)

TLC was carried out in different solvent systems:-

a) 8:2 (hexane - ethyl acetate)

b) 6:3 (hexane - ethyl acetate)
c) 5:5 (hexane - ethyl acetate)
d) 6:2 (methanol - hexane)

After developing all the TLC we got the best result with 5:5 hexane – ethyl acetate solvent
system.

Step 1:

Rf value of Phthalic anhydride-

Distance travelled by the compound

Distance travelled by the solvent

7
 4.5 = 0.89

5.5

Rf value of Phthalimide-

Distance travelled by the compound

Distance travelled by the solvent

5.0

5.5

= 0.90

Step 2:

Rf value of Phthalimide-

Distance travelled by the compound

Distance travelled by the solvent

5.0

5.5

= 0.90

Rf value of 4-Nitrophthalimide-

Distance travelled by the compound

Distance travelled by the solvent

1.5

5.5

= 0.27

8
Visualization of TLC results:

Allow solvent to evaporate from surface of TLC plate. The view results under UV light (Look
the grayish spots on the fluorescent green background). Mark spots with a pencil while viewing
under UV.

APPLICATIONS OF TLC

1. Separation chemistry: mainly for isolation and separation of individual compounds of a

mixture. TLC is more advantageous because almost all chemical substances can be
separated and high speed of separation.

2. Selectivity of TLC is high: very closely related chemical compounds can also be clearly
resolved by adsorption or partition TLC

3. Purification of the sample can be done.

4. Identification of related compounds in the drugs.

5. Separation of mixtures of drugs of chemical or biological origin, plant extracts etc.

6. Separation of carbohydrates, vitamins, antibiotics, glycosides etc.

7. Purification of organic compound: small amount of sample can be purified 30-40 mg of

crude materials can be separated in two 20 cm plates.

8. Examination of reaction: By products identification or estimation can be done.

9. Limitation: TLC is limited to small scale use only.

10. TLC is a quick, inexpensive macroscale technique that can be used to:

11. Determine the number of components in a mixture, verify a substance's identity, monitor
the progress of a reaction, determine appropriate conditions for column chromatography
and analyze the fractions obtained from column chromatography .[1]

9
ADVANTAGES OF TLC
The main advantages of TLC are
1. Requires simple equipment

2. Short development time: TLC takes one hour for development but paper and column
chromatography take several hours to days.

3. Wide choice of stationary phase: Method may be employed for adsorption, partition
(including reverse phase chromatography or ion exchange chromatography).

4. Easy recovery of separated compounds: While removing the Powderly coating of the plates
by scraping with a knife or scrapper, the separated components can be recovered easily. The
spot or zone may be removed quantitatively.

5. Preparation effect: superior than paper chromatography

6. Thickness of the layer: Variable thickness is available from preparative to analytical.

7. Easy visualization of separated compounds: detection of fluorescence compounds under UV

light is easier than on paper because the inorganic background does not fluoresce

8. Sensitivity: 10 to 100 times more than paper chromatography

9. Chemically inert stationary phase.

10
Synthesis of 4- Nitro Phthalimide from Phthalic anhydride

Apparatus use: Round bottom flask, condenser, Beaker, watch glass, filter,chemical
and reagent, funnel etc.

Step I : Synthesis of Phthalimide from Phthalic anhydride

Procedure:

mixture of phthalic anhydride (15g) a urea (3.5g) was taken in a 250 ml beakerand
heated on a sand bath at the temp. of 130 - 35°c.
Froath of solid mass was form with the rise of temperature.
When the reaction subside the spongy solid is obtained.
On cooling some water was added to the spongy solid filtered
Solid was washed with water & dried.
Phthalimide was recrystallised with alcohol.[5]

Crude Phthalimide

Melting Point - 227-228°C

Molecular weight - 11.42 gm

Crystalline Phthalimide

Melting Point - 230°C

Molecular weight - 0.4066 gm

11
SYNTHESIS OF PHTHALIMIDE FROM PHTHALIC ANHYDRIDE

12
Observation:-

Yield of Phthalimide – 11.42 gm

% Yield of Phthalimide – Ex Yield /Theo. Yield ×100 = 11.42 / 15 ×100

= 76.13%

CALCULATION:-

Calculating% yield of Phthalimide

I mole of Phthalic anhydride yields - 1 mole Phthalimide148

g of Phthalic anhydride - 147g of Phthalimide

1g of Phthalic anhydride – 147 / 148

15g of Phthalic anhydride – 147 / 148 ×15 = 14.85 (Theoretical Value)Observed

wt. of Phthalic anhydride - 11.42

Yield % = observed wt. of Phthalic anhydride

Theoretical wt. of Phthalic anhydride X100

11.42
= ×100
14.85

Yield %= 76.90%

Melting Point of crude Phthalimide = 227°C - 228°C

13
Melting Point of crystalline Phthalimide = 230°C

2. Step (II) - Synthesis of 4-nitro Phthalimide from Phthalimide

Procedure:-

• 200 ml beaker containing 41 ml of nitrating mixture (6 ml Nitric acid +35 mlconc.

Sulphuric acid) was cooled. Temperature must be controlled in this reaction
• 5 gram of Phthalimide was rapidly added to the nitrating mixture with stirringwhen
the temp was maintain below 10 °C.
• Precaution was taken that the temperature does not exceed above 10°C whilethe
addition
• After complete addition the ran mixture was kept at room temperature or around1-2
hours.
• Clear pale yellow was poured slowly with vigours strring solution into a 200mlbeaker
containing crust ice .
- Crude 4-nitro Phthalimide was filtered and washed twice with cold water.

- Crude 4-nitro Phthalimide was recrystallised with alcohol.[5]

SYNTHESIS OF 4-Nitro PHTHALIMIDE FROM PHTHALIMIDE

14
 Calculating yield %of crude 4-nitro Phthalimide

1 Mole of Phthalimide gives – 1 mole of 4-nitro Phthalimide

147g of Phthalimide gives - 209 of 4-nitro Phthalimide
5g of Phthalimide gives – 209 / 147 × 5 = 7.1g
Observed wt. of 4-nitro Phthalimide - 7.1 g
% Yield of 4-nitro Phthalimide = observed wt. / Theoretical wt. ×100
6.4 / 7.1 ×100
= 90.00 %

Observation:

Weight of 4-nitro Phthalimide - 6.4 gm

% Yield of crude 4-nitro Phthalimide – 6.4 / 7.1 × 100 = 90.00%

Melting Point of crude 4-nitro Phthalimide – 195°C

Melting Point of crystalline 4-nitro Phthalimide - 197 °C

15
Result:

Step (1): Melting Point of crude Phthalimide = 227°C – 228 °C

Melting Point of crystalline Phthalimide = 230°C

Step (2): Melting Point of crude 4-nitro Phthalimide = 195 °C

Melting Point of crystalline 4-nitro Phthalimide = 197°C

16
 DISCUSSION

Thin-layer chromatography (TLC) may seem like a simple technique, but its importance resonates
across diverse scientific fields. Here's why TLC stands as a cornerstone of chemical analysis:

Simplicity and Speed: TLC's strength lies in its straightforward nature. It requires minimal equipment,
is quick to perform, and provides rapid results. This makes it an invaluable tool for initial screening,
reaction monitoring, and quick qualitative assessments of mixtures.

Versatility and Adaptability: TLC's adaptability is another key advantage. It can be applied to a wide
range of substances, from organic compounds to inorganic ions. The choice of stationary and mobile
phases can be tailored to suit the specific needs of the analysis, making it a highly versatile technique.

Visualization Power: TLC provides a visual snapshot of the components within a mixture. The
separation of compounds into distinct spots on the TLC plate allows for easy identification and
comparison. A variety of visualization techniques, from UV light to chemical reagents, further
enhance its analytical capabilities.

A Stepping Stone to Further Analysis: TLC acts as a guide for more sophisticated separation
techniques. The information obtained from TLC, such as Rf values, can inform the selection of
appropriate conditions for column chromatography, a technique used for larger-scale separation and
purification.

Impact Across Disciplines: TLC's applications extend across diverse fields. It plays a crucial role in
organic synthesis for monitoring reactions and assessing product purity. In pharmaceutical analysis, it
ensures the quality and identity of drug substances. In environmental science, it helps detect and
quantify pollutants. Its importance even reaches into fields like food chemistry and forensic science.

In essence, TLC's simplicity, versatility, and visual power make it an indispensable tool for chemists,
researchers, and analysts across a spectrum of disciplines. It serves as a fundamental technique for
understanding, separating, and analyzing the components of complex mixtures, contributing to
advancements in research, quality control, and our understanding of the world around us.

17
 REFERENCE
* Chemistry LibreTexts: Thin Layer Chromatography:
https://chem.libretexts.org/Ancillary_Materials/Demos_Techniques_and_Experiments/General_Lab_Techniques
/Thin_Layer_Chromatography

* University of Colorado Boulder: Thin Layer Chromatography:

https://orgchem.colorado.edu/Technique/Procedures/TLC/TLC.html

Visualization of Spots on the TLC Plate: https://byjus.com/chemistry/visualization-of-spots-on-the-tlc-plate/

* Sigma-Aldrich: TLC Visualization Reagents: https://www.sigmaaldrich.com/US/en/technical-

documents/technical-article/analytical-chemistry/hplc-uhplc/tlc-visualization-reagents

* Journal of Chromatography A: https://www.sciencedirect.com/journal/journal-of-chromatography-a

* Journal of Planar Chromatography - Modern TLC: https://www.springer.com/journal/10337

18
 HARDNESS OF WATER
INTRODUCTION

The hardness of water is due to the presence of soluble bicarbonates, chlorides and sulfates of calcium
and magnesium. Water which does not give lather with soap is hard water.

Water is the most important compound that is needed for the survival of life on earth. Water is present
in oceans, rivers, ponds, lakes, glaciers, etc. Rainwater is considered pure water because it does not
contain any salt dissolved in it though there are dissolved gases present.

Water can be classified as hard water and soft water.

Soft water: It lathers with soap. Water which is obtained from the rains is soft water. This water is
suitable for household purposes, for example, laundry and cleaning. It is treated water. It is left with
only cations and that is sodium. It has a salty taste.

Hard water: It is known as hard water because of the presence of calcium and magnesium salts.
Hard water does not lather with soap but instead forms a precipitate. The water with naturally present
minerals like magnesium and calcium with detectable amount is called hard water. These minerals
are beneficial for health. They add flavour to hard water.

Difference between Hard and Soft Water

Hard Water Soft Water

It is rich in minerals Contains very few elements
Soap is not so effective Soap is easily effective
No foam and lather from soaps Bubbly lather from soaps
Leaves spots on the washed dishes after Does not leave any spots on dishes after
they are dried they are dried
Contains minerals like magnesium and Contains sodium ion
calcium
Sometimes preferred drinking water Sometimes not preferred drinking water
Example: Groundwater like deep wells Example: Rainwater

19
 What is Hard Water?

Hard water has high mineral content. It is formed when water percolates through the deposits of chalk
and limestone, which are made up of magnesium and calcium carbonates. It does not lather with soap,
so it is not suitable for laundry purposes

The hardness of water is harmful to the boilers as the deposition of salts occurs, which reduces the
efficiency of the boiler. Hard water is safe to drink, but using it over a long interval of time can lead
to many problems like:

1) Strains in skin.
2) Water appliances work harder, resulting in higher water bills
3) Spots appear on clothes and linens

Types of Hardness of Water

Hardness of water can be classified into two types:

1) Temporary Hardness
2) Permanent Hardness

Temporary Hardness of Water

The presence of magnesium and calcium carbonates in water makes it temporarily hard. In this case,
the hardness in water can be removed by boiling the water.

20
When we boil water, the soluble salts of Mg(HCO3)2 are converted to Mg(OH)2, which is insoluble,
and hence gets precipitated and is removed. After filtration, the water we get is soft water

Permanent Hardness of Water

When the soluble salts of magnesium and calcium are present in the form of chlorides and sulphides
in water, we call it permanent hardness because this hardness cannot be removed by boiling.
We can remove this hardness by treating the water with washing soda. Insoluble carbonates are
formed when washing soda reacts with the sulphide and chloride salts of magnesium and calcium,
and thus, hard water is converted to soft water.

Disadvantages of Hardness

1) Wastage of soap
2) Wastage of fuel
3) Formation of scales on metallic boilers.

Remove the Hardness of Water (Temporarily)

By Boiling:

Soluble bicarbonates are converted into insoluble carbonates, which are removed by filtration.
Reactions:

21
 Ca(HCO3)2 → ΔCalo3↓ + H2O + CO2
Mg(HCO3)2 → ΔMgCO3↓ + H2O + CO2

By Clarks Method:
Calcium hydroxide is Clark’s reagent. It removes the hardness of water by converting bicarbonates
into carbonate.
Reaction:
Ca(OH)2 + Ca(HCO3)2 → 2CaCO3↓ + 2H2O
How to Remove Permanent Hardness of Water?

Gan’s Permutit Method:

In this method, sodium aluminium ortho silicate, known as permutit or zeolite, is used to remove the
permanent hardness of the water.
Na2 Al2 Si2 O8.KH2O + Ca++→ 2Na+ + Ca Al2 Si2 O8.xH2O
Calgon’s Process:

In this method, sodium-hexa-meta-phosphate (NaPO3)6, known as Calgon, is used. The hardness in

water is removed by the adsorption of Ca++ and Mg++ ions.
Ion Exchange Resin Method:

In this method, the permanent hardness of water is removed by using resins. Ca++/Mg++ ions are
exchanged with Cl–, and SO4-2 ions are exchanged with anion exchange resin (RNH2OH).
Demineralised water is formed in this process.
⇒ 2RCOOH + Ca++ → (RCOO)2Ca + 2H+
⇒ RNH2OH + Cl– → RNH2Cl + OH–
⇒ H+ + OH– → H2O
Harmful Effects of Hard Water

Some of the most common signs of hard water include the following:
1) Linens and clothes look dull and feel rough.
2) Ugly stains on white porcelain and scale build-up on faucets
3) Low water pressure from showers due to clogged pipes.
4) Chalky, white residue or spots appear on dishes.
5) Strains appear in the shower.

22
 EDTA (Ethylenediamine tetraacetic acid)

The team complexoner was proposed for the compound likes EDTA., nitrilo triacetic acid by
Schwarzenbach who was investigated large number of amino- carboxylic acids of, this type
"Complexons are nothing but complexing agents i.e., they form complexes with various metals. These
complexones co-ordinate with metal ions through several atoms donor at once,
simultaneously forming several chelate rings and giving very stable complexes.
Generally five or six chelate brings are formed which are most stable.
Iminodiacetic acid, HN (CH, COOH), is the simplest possible complex one and all others are regarded
as derivatives of the substance. The most important of these isethylene diamine tetra acetic acid
(EDTA). The use of EDTA as a titrating agent for metal ions has attracted the attention of many
investigators.
Looking to the structure of EDTA it is apparent that in its molecule. In addition to four acidic
hydrogen each nitrogen atom has an unshaped pair of electron. Therefore a molecule of EDTA has six
potential sites for bonding with a metal ion.
Out of all the complexing agents available, ethyl diamine, tetra acetic acid has been widely in the
analysis due to the following reason:
(1) It is available comparatively at a low price, highly soluble in water and acts as primary standard.
(2) It has six ligands atom. (Four acetic hydrogen and two nitrogen atoms) available for bonding with
almost all metal ion.
(3) It forms strainless five memberied rings of chelations resulting in complexes of sufficient stability
to form the basis of volumatic analysis.
(4) It reacts with all cations at least to some extent with the exception of alkalic metals.
(5) The abbrevation HAY, H₂Y, H₂Y2, HX-3 etc. are often used for EDTA and its ions.
(6) The free acid HaY is not often employed for the preparation of standard solution of EDTA
because of its limited solubility likewise the tetra sodium salt Nagy is not suitable owing to its
extensive hydrolysis in sodium ions which makes the solution highly alkaline. The sodium salt, Na₂
H₂ is most useful for analytical work because it can be obtained in high state of purity as the
dehydrate C10H14 Of N2 Na₂ · 2H₂0. Crystallization of the commercial samples gives analytical pure
sodium salt and can be conveniently used as primary standard.
(7) Almost all the complexes are soluble in water.
(8) Martly complexes are colourless. But those of the coloured ions invaribly possess deep colours.
(9) The complexes of divalent metals are stable in ammonical solution, while those of trivalent
transition metals are stable even in strong mineral acid.

23
Reaction of EDTA with metal ions

EDTA in general forms 1:1 complexes with metallic ions.

It is quite clear that the resulting complexes have similar structure but differ
from one another in the charge they carry. It is quite obvious from the above reactions that the
equilibrium is markedly affected the pH of the solution. Welcher has worked out a rough correlation
between charge on the cation, pH of the solution and the stability of the resulting complex.

Structure of a divalent metal EDTA chelate

(a) The complexes of EDTA with divalent metals are very stable in basic

or slightly acidic solutions.

(b) Complexes of the trivalent metals are stable in the pH range 1-2.

(c) Complexes of quadrivalent metals are generally stable at a pH less than one.

TITRATION METHODS USING "EDTA"

The most common procedures employed in the titration of EDTA are described below:

1. Direct Titration Method: When a metal ion is titrated directly with a standard solution of EDTA,
a suitable indicator must be available. Magnesium and number of other divalent ions can be
estimated using the metal. Indicator Eriochrome Black Tis also known as Solochrome Black T. The
titration is carried out in somewhat alkaline solution (pH = 10 by NH, OH).

Mg^2+ + H2Y MgY^2- MgY^-2 + 2H+

24
Eriochrome indicator is a complex organic chelating agent possessing the following formula

The colours at different pH values given by Eriochrome black T are Red …… Below
pH 5.5
Blue ……Between pH range 7-

11 Yellowish-orange …… Above pH 11.5

Between pH range 7-11, when metallic salts are added the color of the indicator changes from blue to
red. At the end-point in EDTA titrations the indicator will be set free and colour change will be form
red to blue.

Back Titration Method: Many metals cannot be titrated directly due to following reasons:

(a) Some metallic ions form stable complexes with EDTA.

(b) Some metallic ions may precipitate from the solution in necessary for the titrations.

(c) Some metals may react with EDTA very slowly.

(d) No suitable metal indicator is available. In such cases and excess of standard EDTA solution
is added and resulting solution if buffered to desired pH and excess of reagent, is titrated with a
standard metal ion solution. A solution of zinc chloride or sulphate, or magnesium n chloride or
sulphate is often used. "The end point is conveniently detected wt (Eriochrome black T in case of
magnesium).

Substitution Method: This method is mainly employed for the titration of calcium with EDTA.
Calcium with Euriochrome black T forms a relatively weak soluble coloured complex which is not
sufficiently stable to serve as an indicator in the titration of calcium with EDTA.

As such a small quantity of magnesium-EDTA chelate is added to the calcium ion before its titration
with EDTA. Since the calcium chelate is far more stable than the magnesium chelate, the following
reaction takes place:

Ca^2+ + MgY^2- CaY^2- + Mg2+

Hence, addition of EDTA to the solution of calcium ion containing a small quantity of magnesium
chelate first leads to the formation of calcium chelate in preference to magnesium chelate. Thus at the
end point the reaction is in between EDTA (H, Y-) and Mg+ ion for which there is a sharp change in
colour, Le, from wine red to blue.

25
 4. Alkalimetric Titration Method: When disodium salt of EDTA i.e., Na2H2 Y is added in metal
ion solution, equivalent amount of H" ions are liberated, which may be titrated with suitable base
in the presence of acid-base indicator.

5. Miscellaneous Method: Certain anions can be determined indirectly EDTA which based on
precipitation or complexation with the selected cations. For example, halides may be determined
by the precipitation with Ag. The silver halide is dissolved in the solution of [Ni(CN)4]^2-
complex ion, which readily liberates Ni* and titrated rapidly with standard solution of EDTA using
murexide indicator.

To estimate the hardness of water (permanent and temporary by complexometric method using
EDTA

Requirement: Sample of water, 0.01M EDTA solution, Erichrome black-T indicator, ammonia buffer
solution.
Theory: EDTA (Ethylenediamine Tetraacetic Acid) is a reagent which form complexes with metals. In
form of its disodium salt. Dissociation constant values indicate that EDTA behave like dicarboxylic
Salt i.e two of its carboxylic group ate strongly acidic and the other two hydrogens are released during
formation of complex.

26
The ionisation of this complexes depends on the PH of the solution. Hence, in this titrations, ptl
sensitive indicates are used. The indicators we cute an azo dye called Erichrome Black-T. This forms a
metal indicator complex, the stability of which is lower than thank of the metal EDTA complex. The
solution is initially red, as the titration proceeds, the metal ions form more stable complex with
EDTA, hence the indicator anion goes in to solution. As the indicator anion
accumulates. the colour change. Water contaminated with Ca add on is called hard water as it
gives hardly any lather with 100p These stales are in trace amounts, hardness is excludes in parts per
million (PPM)
Procedure

1) Pipette out 25ml of sample water in a titration flask add 5ml of buffer solution and 4-5 drops of
indicator-Titrate it against EDTA solution, which is present in burette

2) Al the end point colour change from red to blue. Note the reading reading of burette.

3) Some procedure is repeated with boiled water. We will filter off the water and titrate the filterate
with EDTA for permanent hardness of water.

27
 Calculation

a) Determination of total hardness of water

M1×V1 = M2V2
(KNOWN) = (UNKNOWN)
EDTA = WATER
SAMPLE
M1×B.R =
M2×25
0.0L×12.5 = M2×25

M2 = 0. 01× 1.25
25

M2 = 0.05 (Molarity of water sample)

No. of millimoles in caco3
 MV In 25 ml of water
 0.05

No. of gms of caco3 in 1 litre = 0.05×1000

25
= 0.2
100 × 0.9 = 20
No. of milligram in 100 gm of water
1000 × 20 = 20000 mgl 1000gm

For hot water

M1×V1 = M2×V2
EDTA WATER SAMPLE
0.01 × 7.5 = M2 × 25
M2 = 0.01 × 7.5
25
= 0.0014
No. of millimoles of caco3 = 0.014
No. of gms of caco3 in litre = 0.0014 × 1000
25
= 0.12
100 × 0.12 = 12
No. of miligrams in 10000 gm of water
1000 × 12 = 12000 mgl 1000 gm
Temporary hardness = Total hardness – permanent hardness 20000
-12000
8000mgl 1000gm
Result:
8000mgl1000gm. This is equivalent to total hardness in ppm.
28
Precautions:

1. The apparatus should be cleaned properly with distilled water.

2. Reaction content should be continuously shaken.
3. The PH should be maintained during titration.
4. The end point should be observed correctly.

29
 DISCUSSION

Hard water, a common household nuisance, stems from a high concentration of dissolved minerals,
primarily calcium (Ca²⁺) and magnesium (Mg²⁺) ions. While not a health risk, these minerals can
impact everything from our appliances to the effectiveness of soap. Let's explore the chemistry of hard
water, including its relationship with pH.

The Mineral Makeup of Hard Water

As water travels through soil and rock, it dissolves minerals like calcium carbonate (CaCO₃) and
magnesium sulfate (MgSO₄). These compounds dissociate into their ions:

* CaCO₃ (s) ⇌ Ca²⁺ (aq) + CO₃²⁻ (aq)

* MgSO₄ (s) ⇌ Mg²⁺ (aq) + SO₄²⁻ (aq)

It's these dissolved Ca²⁺ and Mg²⁺ ions that characterize hard water.

pH and its Role

pH, a measure of the acidity or alkalinity of a solution, plays a subtle yet significant role in water
hardness. Here's how:

* Carbonate Chemistry: The carbonate ion (CO₃²⁻) is a weak base. In water, it participates in a series
of equilibria that affect pH:

* CO₃²⁻ (aq) + H₂O (l) ⇌ HCO₃⁻ (aq) + OH⁻ (aq)

* HCO₃⁻ (aq) + H₂O (l) ⇌ H₂CO₃ (aq) + OH⁻ (aq)

* pH Influence on Solubility: Higher pH (more alkaline) favors the formation of carbonate ions
(CO₃²⁻). Since calcium carbonate (CaCO₃) is less soluble than calcium bicarbonate (Ca(HCO₃)₂), a
higher pH can lead to precipitation of CaCO₃, reducing hardness.

30
* Impact on Softening: Softening techniques like adding lime (Ca(OH)₂) raise the pH, promoting the
precipitation of calcium carbonate and reducing hardness.

Temporary vs. Permanent Hardness and pH

The two types of hardness respond differently to pH:

* Temporary Hardness: Caused by bicarbonates, is more sensitive to pH changes. Heating or raising

the pH can readily precipitate out the carbonates.

* Permanent Hardness: Due to sulfates, chlorides, and nitrates, is less affected by pH. Softening
requires methods like ion exchange or reverse osmosis.

The Interplay of Factors

While pH influences hardness, it's not the sole determinant. The concentrations of calcium and
magnesium ions, the presence of other dissolved ions, and the water's temperature all contribute to the
overall hardness.

Understanding the relationship between hardness and pH provides a more nuanced perspective on this
common water quality issue. It highlights how seemingly simple factors like pH can influence the
solubility of minerals and the effectiveness of softening methods, showcasing the intricate chemistry at
play in our everyday lives.

31
 REFERENCE

1. American Water Works Association (AWWA). (2012). Water Quality & Treatment: A Handbook on
Drinking Water (6th ed.). McGraw-Hill.
2. Water Quality Association (WQA): [https://goldbook.iupac.org/]
3. United States Geological Survey (USGS) Water Science School: [https://www.usgs.gov/special-
topics/water-science-school/science]
4. Environmental Protection Agency (EPA) Drinking Water Requirements:
[https://www.epa.gov/ground-water-and-water/national-primary-water-regulations]
5. International Union of Pure and Applied Chemistry (IUPAC) Gold Book: [https://goldbook.iupac.org/]

32
 pH OF BUFFER SOLUTIONS

INTRODUCTION

pH
The pH scale ranges from 0 to 14, with 7 denoting neutrality. Solutions with a pH below 7 are acidic,
while those above 7 are alkaline. The pH of water is intricately tied to the balance between hydrogen ions
(H+) and hydroxide ions (OH-) in the solution. A lower pH signifies a higher concentration of hydrogen
ions, indicating greater acidity, while a higher pH reflects a higher concentration of hydroxide ions,
signaling alkalinity.

The pH of water holds significant implications across various domains, including environmental science,
industrial processes, and public health. In the realm of drinking water quality, maintaining an optimal pH
level is paramount to ensure the safety and palatability of water for human consumption. Acidic water can
leach metals from plumbing systems, leading to potential contamination, while alkaline water can cause
scaling issues that compromise infrastructure integrity.

Environmental monitoring relies heavily on pH measurements to assess the health of aquatic ecosystems.
Fluctuations in pH levels can have detrimental effects on aquatic life, as many species are sensitive to
changes in acidity or alkalinity. Acidification or alkalization of water bodies can disrupt ecological balance
and biodiversity, underscoring the importance of monitoring and regulating pH in natural water sources.[1]

Industrial processes also heavily rely on precise pH control to optimize chemical reactions, prevent
corrosion of equipment, and ensure product quality. Maintaining specific pH ranges is critical for the
efficiency and effectiveness of various industrial operations, with deviations potentially leading to
production inefficiencies and product defects. Accurate monitoring and adjustment of water pH are
essential for maintaining operational efficiency and regulatory compliance in industrial settings.

A variety of methods are available for measuring and adjusting water pH, including sophisticated pH
meters that provide real-time data for precise control of treatment processes. Colorimetric indicators offer a
rapid visual estimation of pH based on color changes induced by specific chemicals. Chemical additives
such as acids or bases can be employed to adjust water pH to desired levels for specific applications,
ensuring optimal performance and product quality.

33
In conclusion, a comprehensive understanding of pH is indispensable for safeguarding water quality,
environmental sustainability, and industrial productivity. By effectively monitoring and managing water
pH levels, individuals and industries can uphold the integrity, efficiency, and sustainability of water
resources across diverse applications and sectors. pH solutions are solutions that have been adjusted to a
specific pH level using acidic or alkaline substances. The pH scale measures the acidity or alkalinity of a
solution, with values ranging from 0 to 14. A pH of 7 is considered neutral, while values below 7 indicate
acidity and values above 7 indicate alkalinity.

pH solutions are commonly used in various applications, including:[2]

1. Laboratory experiments: pH solutions are often used in scientific research and experiments to control
the acidity or alkalinity of a solution. This is important in fields such as chemistry, biology, and
environmental science.

2. Water treatment: pH solutions are used in water treatment processes to adjust the pH of water to meet
specific standards. This helps to ensure that the water is safe for consumption and other uses.

3. Industrial processes: Many industrial processes require specific pH levels for optimal performance. pH
solutions are used to adjust the pH of process fluids to meet these requirements.

4. Agriculture: pH solutions are used in agriculture to adjust the pH of soil or irrigation water. This is
important for ensuring that plants have access to the nutrients they need for healthy growth.

pH solutions can be prepared by mixing acids or bases with water to achieve the desired pH level.
Common substances used to prepare pH solutions include hydrochloric acid, sulfuric acid, sodium
hydroxide, and potassium hydroxide. It is important to handle these substances with care, as they can be
corrosive and potentially harmful if not used properly.

Overall, pH solutions play a crucial role in various industries and applications, helping to maintain the
proper pH levels required for optimal performance and safety.

pH Scales

The pH scale is a fundamental tool in chemistry and various other fields for measuring the acidity or
alkalinity (basicity) of a solution. Essentially, it tells us about the concentration of hydrogen ions (H+)
present in a solution.

Here's a breakdown of the key points:

34
Range and Values:

* The pH scale typically ranges from 0 to 14.

* A pH of 7 is considered neutral, indicating a balance between H+ and OH- ions (e.g., pure water).

* Values below 7 indicate acidity, with lower values representing stronger acids (e.g., lemon juice,
stomach acid).

* Values above 7 indicate alkalinity (basicity), with higher values representing stronger bases (e.g., baking
soda, bleach).

Logarithmic Nature:

* The pH scale is logarithmic, meaning that each unit change represents a tenfold difference in H+
concentration.

* For example, a solution with a pH of 4 is ten times more acidic than a solution with a pH of
5.Calculating pH:

* pH is calculated using the formula: pH = -log[H+], where [H+] represents the molar concentration of
hydrogen ions.

35
BUFFER SOLUTION

Buffer solutions typically consist of a weak acid and its conjugate base, or a weak base and its conjugate
acid. These components work together to resist changes in pH by neutralizing the added acid or base
through a chemical reaction. The weak acid or base component of the buffer reacts with the added acid or
base, preventing significant changes in the overall pH of the solution.

One of the key characteristics of buffer solutions is their ability to resist changes in pH within a specific
range, known as the buffer capacity. The buffer capacity is determined by the concentration of the weak
acid and its conjugate base (or weak base and its conjugate acid) in the solution. Higher concentrations of
these components result in higher buffer capacity, allowing the buffer solution to effectively resist changes
in pH.

Buffer solutions are widely used in various applications, including:[4]

1. Biological and biochemical research: Buffer solutions are commonly used in biological and biochemical
experiments to maintain a stable pH, which is essential for the proper functioning of enzymes, proteins,
and other biological molecules.

2. Pharmaceutical industry: Buffer solutions play a crucial role in pharmaceutical manufacturing processes,
where maintaining a specific pH is necessary for the stability and effectiveness of drugs.

3. Analytical chemistry: Buffer solutions are used in analytical chemistry techniques such as
chromatography and electrophoresis to control the pH of the mobile phase and ensure accurate and
reproducible results.

4. Water treatment: Buffer solutions are used in water treatment processes to stabilize the pH of water and
prevent fluctuations that could lead to corrosion or scaling in water distribution systems.

36
Preparing a buffer solution involves mixing precise amounts of a weak acid and its conjugate base (or
weak base and its conjugate acid) to achieve the desired pH. Common buffer systems include acetic
acid/sodium acetate, citric acid/sodium citrate, and phosphate buffers.

In conclusion, buffer solutions are essential tools in various industries and scientific disciplines where
maintaining a stable pH is critical. Their ability to resist changes in pH makes them invaluable for ensuring
the success and accuracy of numerous processes and experiments.

Key Points[5]

1. pH Range: Buffer solutions are most effective at maintaining a relatively constant pH within a specific
range known as the buffering range. This range typically falls within one pH unit of the pKa of the weak
acid component of the buffer system. For example, if acetic acid (CH3COOH) is used as the weak acid in
a buffer system, the buffering range would be around pH 4.76, which is the pKa of acetic acid.

2. Buffer Capacity: Buffer capacity refers to the amount of acid or base that a buffer solution can neutralize
without significant changes in pH. The buffer capacity is influenced by the concentrations of the weak acid
and its conjugate base (or weak base and its conjugate acid) in the solution. Higher concentrations of these
components result in a higher buffer capacity.

3. Types of Buffer Solutions: There are two main types of buffer solutions: acidic buffers and basic
buffers. Acidic buffers consist of a weak acid and its conjugate base, while basic buffers consist of a weak
base and its conjugate acid. The choice of buffer system depends on the desired pH range and the specific
requirements of the application.[6]

5. Applications: In addition to the applications mentioned earlier, buffer solutions are also used in various
other fields, including food and beverage industry, environmental science, and biotechnology. They are

37
essential for maintaining optimal conditions for chemical reactions, biological processes, and analytical
measurements.

6. Buffer Preparation: To prepare a buffer solution, you need to calculate the amounts of the weak acid and
its conjugate base (or weak base and its conjugate acid) required to achieve the desired pH. This involves
considering factors such as pKa values, concentrations, and volume of the final buffer solution.

Overall, buffer solutions are versatile and indispensable tools in science and industry for maintaining pH
stability and ensuring the success of a wide range of processes and experiments. Their unique ability to
resist changes in pH makes them essential for numerous applications where precise pH control is crucial.[7]

38
 EXPERIMENT
Requirements! NaOH, Oxalic acid, Acetic acid, Sodium acetate, phenolphthalein, pH meter

Preparation of NaOH and Oxalic acid

1- for 1N NaOH (Unknown)

W= NEV/ 1000 = 1x 40 x 250 /1000 -= 10 gm

2- for IN Oxalic acid

W= NEV/ 1000=1 X 63 X 250 / 1000= 15.75 gm

Standardization of NaOH

NaOH is titrated against Oxalic acid with the help of phenolphthalein.

Procedure:-

fill the burette with NaOH. Pipette out 10 ml of the Oxalic acid Solution into a Conical Hask. Add 2-3
drops of phenolphthalein as an indicator in it. Now add slowly NaOH Solution from flask Constantly
burette, shaking the till a pink Colour is just obtained. Note the burette reading and repeat the tetration
until two concordent readings the are obtained.

Observation Table :-

39
So , N1V1 (oxalic acid) =N2V2(NaOH)

1x10=N2* 8.5

1x10=N2 *8.5

N2=10/8.5=1.17

N2=1.17

So, Exact normality of NaOH is 1.17N

Now ,Dilute the 1.17N NaOH Solution to make it 0.5N NaOH solution

Dilution-

N1V1=N2V2

1.17 *V1=(1/2) x2 50

V1=(250/2*1.17) =106.83ml

Prepration of N/2 NaOH-

Use 106.83ml of 1.17N NaoH and make it upto 250ml.

Prepration of Acetic acid –

For 0.5N Acetic acid

Since given acetic acid =17N

N1V1=N2V2

17*V1=0.5*250

V1=(0.5*250) /17 =7.352ml

V1= 7.352ml

Use 7.352ml of given 17N acetic acid and make it upto 250 ml.

Standardization of Acetic acid (CH3COOH)-

Titrate the acetic acid with the help of NaOH using the ionidated phenolphthalein.

Now,

N1V1 (CH3COOH) =N2V2(NaOH)

N1*10=9.1*0.5

40
N1=9.1/20=0.455N

So, Exact normality of CH3COOH is 0.455N

Preparation of 0.2N CH3OOH -

Dilute the 0.455 N CH 3 COOH to make the 0.2 N CH3OOH Sol".

N_{1}*V_{1} = N_{2}*V_{2}

0.455 x V, 0-2 x 250 V_{1} = (0.2 * 250)/(0 * 455) = 50/(0 * 455) = 109.89ml

Use 109.89 ml of upto 250 ml. 0-455 N acetic acid S*theta * l ^ n and make it

Preparation of CH3COONa (0.2N):-

w = (NEV)/1000 = (0.2 * 82 * 250)/1000 = 4.1gm

Preparation of Acetate Buffer-

Result-

pH of I buffer – 4.99

pH of II buffer – 4.63

pH of III buffer – 4.05

Verification of pH with Henderson – Hasselbatch Equation

pH= pKa + log salt/acid

Ka= 1.8*10^5(for acetic acid)

pKa= -log Ka

pKa= -log(1.8*10^-5)

41
pKa= -log(1.8) –log 10^-5

=-0.255+5=4.545

pKa=4.545

1) For I buffer(6:14)

(6mlCH3COOH+14ml CH3COONa)

pH=pKa+log [CH3COONa/ CH3COOH)

pH=4.545+log [(14*0.2)*(6*0.2)

pH=4.545+log7-log3

pH=4.545+0.845+0.477

pH=4.913

2) For II buffer (10:10)

(10mlCH3COOH+10ml CH3COONa)

pH=pKa+log [CH3COONa/ CH3COOH)

pH=4.545+log [(10*0.2)*(10*0.2)

pH=4.545+log1

pH=4.545

3) For I buffer(6:14)

(16mlCH3COOH+4ml CH3COONa)

pH=pKa+log [CH3COONa/ CH3COOH)

pH=4.545+log [(4*0.2)*(16*0.2)

pH=4.545+log1-log4

pH=4.545+0-0.602

pH=3.94

42
 DISCUSSION

* Clear and Concise Explanation of pH: As we effectively defines pH, explains its relationship to acidity
and alkalinity, and describes the pH scale with its range and neutrality point. [8]

* Importance of pH in Various Domains: As emphasizes the significance of pH in different areas:

* Drinking Water Quality: Maintaining optimal pH for safety, preventing metal leaching and scaling.

* Environmental Science: Monitoring pH in aquatic ecosystems to assess health and protect

biodiversity.

* Industrial Processes: Controlling pH for optimizing reactions, preventing corrosion, and ensuring
product quality.

* Methods for Measuring and Adjusting pH: As we outlines various methods, including pH meters,
colorimetric indicators, and chemical additives, providing a comprehensive overview of available tools.

* pH Solutions and Their Applications: As expands on the use of pH solutions in various applications,
including:

* Laboratory Experiments: Controlling the acidity or alkalinity of solutions for research purposes.

* Water Treatment: Adjusting pH to meet safety standards for consumption and other uses.

* Industrial Processes: Maintaining specific pH levels for optimal performance and product quality.

* Agriculture: Adjusting soil and irrigation water pH for optimal plant growth.

* Comprehensive Conclusion: The conclusion effectively summarizes the importance of understanding

and managing pH for water quality, environmental sustainability, and industrial productivity.]

* Introduction to Buffers: Buffer solutions are crucial in maintaining a stable pH despite the addition of
acids or bases. They consist of a weak acid and its conjugate base (or a weak base and its conjugate acid)
working together to neutralize added H+ or OH- ions.

* Mechanism of Action: Buffers resist pH changes by shifting the equilibrium between the weak acid/base
and its conjugate to absorb the added ions. This helps maintain a relatively constant pH within a specific
range.

43
* Examples of Buffers: The bicarbonate buffer system in human blood plays a vital role in maintaining
blood pH near 7.4. Other examples include phosphate buffers in cells and acetate buffers in various
industrial applications.

Specific pH Examples:

* Common Beverages: Provide a table showcasing the pH values of common beverages such as coffee
(around 5), orange juice (around 3.5), and soda (around 2.5), highlighting the acidic nature of many
popular drinks.

* Household Products: Mention the pH of common household products like bleach (highly alkaline,
around 12.6), vinegar (acidic, around 2.4), and baking soda (slightly alkaline, around 8.3), emphasizing the
importance of proper handling and storage.

* Water Sources: Discuss the pH variation in different water sources, such as rainwater (slightly acidic
due to dissolved CO2), seawater (slightly alkaline), and freshwater bodies, which can vary depending on
factors like geology and pollution.

pH and Human Health:

* Digestion: Explain how stomach acid (pH around 1.5-3.5) aids in protein digestion and kills ingested
bacteria. Also, mention the role of pancreatic juices (pH around 8) in neutralizing stomach acid and
facilitating further digestion in the small intestine.

* Enzyme Activity: Discuss how enzymes, crucial for various biochemical reactions in the body, have
optimal pH ranges for their activity. Deviation from these ranges can affect enzyme function and lead to
health problems.

Environmental Impacts of pH Imbalance:

* Acid Rain: Expand on how acid rain, formed by the reaction of sulfur dioxide and nitrogen oxides with
water in the atmosphere, can damage ecosystems, acidify lakes and rivers, and harm aquatic life.

* Ocean Acidification: Discuss how the absorption of excess atmospheric CO2 by oceans leads to a
decrease in pH, impacting marine organisms, coral reefs, and the entire marine food web.

Advanced pH Measurement Techniques:

44
 REFERENCE

1. Harris, D. C. (2010). Quantitative Chemical Analysis (8th ed.). W. H. Freeman and Company.
2. Atkins, P. W., & de Paula, J. (2014). Atkins' Physical Chemistry (10th ed.). Oxford University Press.
3. Housecroft, C. E., & Sharpe, A. G. (2012). Inorganic Chemistry (4th ed.). Pearson Education.
4. Bates, R. G. (1973). Determination of pH: Theory and Practice. John Wiley & Sons.
5. National Institute of Standards and Technology (NIST) pH Measurement:
[https://www.nist.gov/programs-projects/ph-
metrology#:~:text=The%20suite%20of%20NIST%20pH,industry%2C%20government%2C%20and%
20academia.]
6. United States Geological Survey (USGS) Water Science School: [https://www.usgs.gov/special-
topics/water-science-school/science]
7. American Chemical Society (ACS) Chemistry for Life: [https://www.acs.org/]
8. Sigma-Aldrich Buffer Reference Center: [Insert Link Here]
9. Bio-Rad Laboratories pH and Buffers Guide: [Insert Link Here]

45
 POLARIMETER
INTRODUCTION
Polarimeters are instruments that are used to measure the optical activity of substances,
mainly chiral molecules. A polarimeter is mainly used in determining the identity of
molecules, because it is a non-destructive method of identification. It does not require any
chemical reaction to be performed during the identification process. Polarimeters have a
number of functions, from the identification of molecules to measuring the rate of chemical
reactions. In the chemical industry, polarimeters are used to measure the purity of substances
based on their optical properties and are also used in the determination of substances in
mixtures. In the food industry, polarimeters are used to distinguish between different types of
nutrients, drugs, steroids, vitamins, and carbohydrates amongst other substances. In the sugar
industry, quality control is done through the use of a polarimeter. Inthe pharmaceutical
industry, a polarimeter is used to monitor the quality and purity. As far as biological
applications are concerned, polarimeters are used in the identification and
purity of monomers of carbohydrates, lipids, nucleic acids, and proteins, based on their
optical properties. The polarimeter was invented by Étienne-Louis Malus, who also
discovered the Malus’ law, also known as Malus law of Polarimetry. Malus’ law states that
when plane polarized light is incident to an analyzer (that is the plane of polarized light is
90o to an analyzer), the intensity of light transmitted by the analyzer is directly proportional
to the square to the cosine of the angle between polarizer and the plane of light.[7]

Therefore, according to Malus,

I α cos2 θ

Therefore, I= Io Cos2 θ

According to Malus’ law, the polarizer and the analyzer are supposed to be ideal, and hence,
there would be only plane polarized light passing through the analyzer. Instrument measuresthe
rotation of polarized light as it passes through an optically active substance and the
tendency of the molecule to rotate the plane polarized light towards clockwise or anti-
clockwise direction whose extent of the rotation can be measured. In principle, a pair of
crossed polarizers (a pair with their pass axes perpendicular to each other) may be used as
polarimeter. No light will emerge from such a combination. If an optically active substance is
introduced between them, the plane of polarization of the light emerging from it may be
rotated by a certain angle (let it be) and the second polarizer will not be able to block the
light now.
The second polarizer will have to be rotated by an angle in the same sense to make the fieldof
view dark again. The angle of rotation can thus be measured by fitting a circular scale to
the second polarizer.

46
Optical Activity
The ordinary lights are composed of rays of different wavelengths vibrating in all directions
perpendicular to the path of its propagation. These vibrations can be made to occur in a
single plane by passing ordinary light through the polarizing Nicol prism. Such light whose
vibrations occur in only one plane is called plane polarized lights. A beam of unpolarized
light is transformed to plane-polarized light by passing it through a polarizing filter. This filter
is usually a Nicol prism made up of calcite. This plane polarized light now passes through the
sample tube containing the substance to be examined. The sample is optically active if it
rotates the plane of polarized light. The direction and magnitude of rotation are measured
using a second polarizing filter (the analyzer) and is given as α, the observed rotation. A
substance that does not rotate the plane of polarized light is said to be optically inactive. All
achiral substances are optically inactive. Rotation of plane polarized light in the clockwise
direction is called dextrorotatory (+), and rotation in the anticlockwise sense is called
levorotatory (-) rotation. Optical activity is given in terms of specific rotation [α].
Specific rotation is calculated from the observed rotation according to the expression.[10][

]= α/cxl where c – concentration in g/mL and l – length of the polarimeter

tube in dm.

For a compound to be optically active, it should not possess plane of symmetry, centre of
symmetry and improper axis of rotation. Presence of proper axis of symmetry is not a
required condition for optical activity. Optically active compounds may or may not contain

47
Cn. Chirality is not a necessary condition for optical activity since there are molecules that
do not possess a chiral center and yet display optical activity. Absence of symmetry or the
three symmetry elements mentioned above is a necessary condition for optical activity.
Optical rotation, also known as polarization rotation or circular birefringence, is the rotationof
the orientation of the plane of polarization about the optical axis of linearly polarized lightas it
travels through certain materials. Circular birefringence and circular dichroism are the
manifestations of optical activity. Optical activity occurs only in chiral materials, those
lacking microscopic mirror symmetry. Unlike other sources of birefringence which alter a
beam's state of polarization, optical activity can be observed in fluids. This can include gasesor
solutions of chiral molecules such as sugars, molecules with helical secondary structure such
as some proteins, and also chiral liquid crystals. It can also be observed in chiral solids such
as certain crystals with a rotation between adjacent crystal planes (such as quartz) or
metamaterials. Compounds which rotate the plane of polarized light are called optically active
compounds and this property is known as optical activity. Rotation of plane of
polarized light can be of two types :

• Dextrorotatory : If the compound rotates the plane of polarization to the

right(clockwise) it is said to be dextrorotatory (Latin: dexter-right) and is denoted by (+), or
‘d’.

• Levorotatory : If the compound rotates the plane of polarization to the

left(anticlockwise) it is said to be levorotatory (Latin: laevus-left) and is denoted by (-) or ‘l’.

The change in the angle of plane of polarization is known as optical rotation. The optical
rotation is detected and measured by an instrument called polarimeter. The measurementof
optical activity is reported in terms of specific rotation [α], which is given as , [α] λ t =
α/l×c

[α]= specific rotation t = temperature of

measurement λ=wavelength of the light used α=

observed angle of rotation l= length of sample

tube in decimeter c=concentration of the samplein

g/mL of solution

48
Types of polarimeter
• Laurent's half-shade polarimeter

• Biquartz polarimeter

• Manual.

• Semi-automatic

• Fully automatic

Laurent’s Half Shade Polarimeter:

Laurent’s half-shade polarimeter consists of a semi-circular half wave plate ABC of quartz
(cut parallel to optic axis) so that it introduces a phase change of between the extraordinary
and the ordinary rays passing through it, and a semi-circular glass plate ADC. The thickness
of the glass plate is such that it absorbs the same amount of light as the quartz

plate. Let the plane a vibration of the plane polarized light incident normally on the half
shade device be along PQ making an angle with AC. The vibrations emerge from the glass
plate part of the half shade device as such i.e., there is no change along the plane PQ. Insidethe
quartz plate, which is doubly refracting, the light is divided into Inside the quartz plate, which
is doubly refracting, the light is divided into two components as we know, one
ordinary component XX’ and the other extraordinary component parallel to the optic axis
along YY '.[4]

The two components travel along the same direction through separation but with different
velocities. The ordinary component moves with greater velocity than the extraordinary
component. On emergence, a phase difference of is introduced between them. Due to this
phase difference the direction of the ordinary component gets reversed. If the initial
position of the ordinary component is represented by OM, then the final position is
represented by ON. Now, on emergence the resultant of the extraordinary.

49
OL and ordinary component ON will be OR making an angle theta with the y axis. The
vibrations of the beam emerging out of the quartz portion of the half shade device will be
along RS. That is the change. The essential parts of this polarimeter are a monochromatic
light source, a convex lens which changes the incident light beam into a parallel one. A
polarization which makes this light plane polarized then, the Laurent’s half shade device and
then a tube containing the optically active experimental substance. The light beam emerging
from this cube passes through an analyzer. This analyzer is capable of rotation about a
common axis. The rotation of the analyzer can be read on a circular scale fitted with
verniers. The light after passing through the analyzer is viewed through a telescope which is
focused on the half shade device. If the pass direction directions of the analyzing polarizer
which is capable of being rotated and if it is fitted with the circular scale. When the past
direction is parallel to PQ then light from the glass portion will pass unobstructed while the
light from the quartz portion will be partly obstructed. Due to this, the last half will appear
brighter than the quartz half.

On the other hand, if the pass direction of the analyzer is parallel to RS, light from the quartz
portion will pass unobstructed, but the light from the glass portion will be partly obstructed.
Thus, the quartz half will appear brighter than the glass half. If however, the past direction of
the analyzer is parallel to AC, y axis it equally inclined to the two planes polarized lights.
Hence the field of view, in the two halves, will be equally bright because the half shade
device serves the purpose of dividing the field of view in 2 halves. A little change in the
direction of the pass direction of the analyzer makes one half brighter, the other half darker
focused on the half shade device and analyzer is rotated till the 2 halves are equally bright.
This position is noted on the circular scale. The tube is then filled with the optically active
solution and placed in position. The analyzer is rotated, and it is brought to a position such
that the two halves are equally bright again. This new position is noted and the difference
between the two regions gives the angle of rotation pretty accurately.

Biquartz Polarimeter :

It consists of a white light source, a convex lens as before, which renders the light into a
parallel beam, the polarizer changes the incident beam into a plane polarized beam, then a
biquartz plate, then the experimental tube containing the active substance and a polarizer
working as an analyzer as before. A telescope is fitted with a circular scale and is focused on
the biquartz plate. Biquartz plate consists of two semicircular plates of quartz. One left-
handed quartz L and the other right-handed quartz R, each of thickness about 3.75
millimeters. Both are cut perpendicular to the optic axis.[10]

50
This means that propagation here is along the optic axis now. They are joined together along
the diameter PQ. When the plane polarized white light passes through a biquartz plate
normally, along the optic axis the phenomenon of rotary dispersion occurs because the
planes of vibrations of different colors are rotated through different angles. Remember, we
have seen that the amount of rotation is proportional to 1 upon lambda square and rotation will
be in one sense for the left-handed portion and other direction for the right-handed portion.
The amount of rotation is maximum for violet, which has the minimum wavelengthand least
for red. The sense of rotation is opposite in the two halves. The amount of rotationalso
depends on the thickness. For a thickness of 3.75 millimeters, millimeters, the rotation of the
plane of polarization for yellow light is about 90 degrees. Hence YOY is a straight line.
If the pass direction of the analyzer is parallel to POQ, the yellow light will not be
transmitted through the analyzer, Malus Law and the appearance of two halves will be
similar. The two halves will have a grayish violet tint called the tint of the passage. When the
analyzer is rotated to one side from this position, one half of the field of view appears blue,
while the other half appears red. When the analyzer is rotated in the opposite direction, the
colors are interchange the first half which was bluish earlier now appears red and the second
half which was reddish earlier now appears blue. The position dependence of the tint of
passage is very sensitive and is used for accurate determination of the angle of optical
rotation.

Manual
The earliest polarimeters, which date back to the 1830s, required the user to physically
rotate one polarizing element (the analyzer) whilst viewing through another static element
(the detector). The detector was positioned at the opposite end of a tube containing the
optically active sample, and the user used his/her eye to judge the "alignment" when least
light was observed. The angle of rotation was then read from a simple fixed to the moving

51
polarizer to within a degree or so. Although most manual polarimeters produced today still
adopt this basic principle, the many developments applied to the original opto- mechanical
design over the years have significantly improved measurement performance. The
introduction of a half-wave plate increased "distinction sensitivity", whilst a precision glass
scale with vernier drum facilitated the final reading to within ca. ±0.05°. Most modern
manual polarimeters also incorporate a long-life yellow LED in place of the more costly
sodium arc lamp as a light source.

Semi-automatic
Today, semi-automatic polarimeters are available. The operator views the image via a digital
display adjusts the analyzer angle with electronic controls.

Fully automatic polarimeter Fully automatic polarimeters are now widely used and simply
require the user to press a button and wait for a digital readout. Fast automatic digital
polarimeters yield an accurate result within a few seconds, regardless of the rotation angle ofthe
sample. In addition, they provide continuous measurement, facilitating High-performanceliquid
chromatography and other kinetic investigations. Another feature of modern polarimeters is the
Faraday modulator. The Faraday modulator creates an alternating currentmagnetic field. It
oscillates the plane of polarization to enhance the detection accuracy by allowing the point of
maximal darkness to be passed through again and again and thus be determined with even more
accuracy. As the temperature of the sample has a significant influence on the optical rotation
of the sample, modern polarimeters have already included Peltier elements to actively control
the temperature. Special techniques as temperature controlled sample tubes reduce measuring
errors and ease operation. Results can directly betransferred to computers or networks for
automatic processing. Traditionally, accurate fillingof the sample cell had to be checked outside
the instrument, as an appropriate control from within the device was not possible. Nowadays a
camera system can help to monitor the sample and accurate filling conditions in the sample cell.
Furthermore, features for automaticfilling introduced by few companies are available on the
market. When working with caustic chemicals, acids, and bases it can be beneficial to not load
the polarimeter cell by hand. Bothof these options help to avoid potential errors caused by
bubbles or particles.

Sources of errors

The angle of rotation of an optically active substance can be affected by:

• Concentration of the sample

• Wavelength of light passing through the sample (generally, angle of rotation and wavelength
tend to be inversely proportional)

• Temperature of the sample (generally the two are directly proportional)

52
• Length of the sample cell (input by the user into most automatic polarimeters to ensure better
accuracy)

• Filling conditions (bubbles, temperature and concentration gradients) Most modern

polarimeters have methods for compensating or/and controlling these errors.[10]

Application:

• To determine product purity by measuring specific rotation and optical rotation of amino
acids, amino sugars, antibiotics, dextrose, steroids, cocaine, diuretics, tranquilizers,
analgesics, codeine, serums, vitamins, etc.

• To measure the ratio of enantiomers in solutions.

• Many optically active chemicals such as tartaric acid, are stereoisomers, a polarimeter can be
used to identify which isomer is present in a sample.

• With a known concentration of a sample, polarimetry may also be applied to determine the
specific rotation (a physical property) when characterizing a new substance.

• Polarimetric method is a simple and accurate means for determination of structure in micro
analysis of expensive and non-duplicable samples.

• It is employed in quality control, process control and research in the pharmaceutical,

chemical, essential oil, Flavors and food industries.

• It is so well established that the United States Pharmacopoeia and the Food & Drug
Administration include polarimetric specifications for numerous substances.[7]

53
To determine the specific rotation of cane sugar solution by polarimeter.

Requirement – Polarimeter, sodium vapour, arc lamp, sugar.

Theory-
Polarimeter

An instrument used to measure optical rotation is called a polarimeter.It

consists of the following basic parts:

1. A light source is usually a sodium vapour lamp.

2.A polarizer (Nicol prism fixed)

1. Ananalyzer (Nicol prism can be rotated on a graduated circular scales).

2. A circular scale to measure the angle of rotation.

3. Tubes to hold the sample solution.

4. An eye piece to view the emergent lights.

54
Measurement of Optical rotation
The tube is filled with distilled water analyzer is turned on a circular graduated scale to
obtain total darkness and the reading on the scale is noted. The tube containing a solutionof
an optically active material is then place polarizes and analyzes On looking through the
analyzer, the field has rotated the plane of polarizer. The analyzer is again turned to obtain
the darkness, and the angle through which it has been turned to found out from the two
scale reading to this angle is equal to the angle of rotation of the given solution. [2]

P -polarizer

L-Light

source C-

Collimating

H-Half-shed prism

T-Polarimeter tube

A- Analyzer

Ep-Eye

pieceE-Eye

The problem in the above measurement is the difficulty in finding out when the view a
exactly as dark as it was without the solution. In order to solve this problem, a small Nicol
prism is placed in between the polarizer and light source. The view is now divided into two
halves whose brightness can be more conveniently compared. On rotating the analyzer,
different position .
B= Balance point

55
The balance point is first observed with distilled water in the tube, by rotating the analyzer on
the circular graduated scale. The scale reading is noted. The tube containing the sample
solution is then placed between the polarizer and analyzer. Analyzer is again rotated to obtain
the balance point. The scale reading again recorded the difference between the two readings
gives the optical rotation of the solution. Specific rotation is given by reading : [α ]1
=100α / l×c [2]

Procedure – Prepare the solution of given substances at three different concentration and
determine the angle of rotation by the position of the analyzer. Filling polarimeter tube with
distilled water and then different solution

Observation -
S.NO % Of Reading of Reading of Angle of Specific
solutions the analyzer the analyzer rotations rotation
with with
distilled solutions [[α]=100α/l×c]
water
(a) (b) (a-b)
1 5% 276° 209° 67° 67°
2 10% 276° 156° 120° 60°
3 15% 276° 146° 130° 65°

CALCULATION –
5% of solutions specific rotation

[α] =100α /l×c

100×67/20×5

= 67 °

10% of solutions specific rotation

[α] = 100 α

/l×c

100×120/20×10

= 60°

56
15%of solutions specific rotation

[α] =100 α /l×c

100×130 / 20×15

= 65°

Result -

Specific rotation of the given substances of the three different concentration ( 5% , 10% ,15%
) 67°,60°,65° .

57
 DISCUSSION

The polarimeter, an essential tool in chemistry and beyond, unlocks the secrets hidden within polarized
light. This instrument quantifies the rotation of plane-polarized light as it interacts with chiral molecules,
providing valuable insights into their structure and concentration. Let's delve into the fascinating world of
polarimetry:

The Fundamentals of Polarized Light:

* Ordinary light: Vibrates in all directions perpendicular to its direction of travel.

* Plane-polarized light: Vibrates in a single plane, achieved by passing ordinary light through a polarizing
filter (like a Nicol prism or Polaroid filter).

The Heart of the Polarimeter:

A polarimeter typically comprises:

1. Light source: Often a monochromatic source, like a sodium lamp.

2. Polarizer: Creates plane-polarized light from the source.

3. Sample tube: Holds the optically active substance being analyzed.

4. Analyzer: A second polarizer, rotatable, used to measure the angle of rotation.

5. Detector: Measures the intensity of light passing through the analyzer.

Optically Active Substances:

* Chiral molecules: Possess a non-superimposable mirror image, like our left and right hands.

* Enantiomers: Pairs of chiral molecules that are mirror images of each other. They rotate plane-polarized
light in opposite directions.

The Magic of Rotation:

* When plane-polarized light passes through a solution of a chiral molecule, the molecule interacts with
the light, causing its plane of polarization to rotate.

* The degree of rotation is specific to the molecule and its concentration.

* Dextrorotatory (+): Rotates light clockwise.

* Levorotatory (-): Rotates light counterclockwise.

Quantifying the Rotation:

58
* Observed rotation (α): The angle in degrees by which the analyzer must be rotated to achieve maximum
light transmission.

* Specific rotation ([α]): A standardized value, independent of concentration and path length:

* [α] = α / (c * l)

where:

* α = observed rotation

* c = concentration (g/mL)

* l = path length (dm)

Applications of Polarimetry:

* Structure determination: Distinguishing between enantiomers and determining their purity.

* Quantitative analysis: Measuring the concentration of optically active substances.

* Reaction kinetics: Monitoring the progress of reactions involving chiral molecules.

* Food industry: Assessing sugar concentrations and detecting adulteration.

* Pharmaceuticals: Ensuring drug purity and identifying counterfeit medications.

Beyond Chemistry:

Polarimetry extends beyond chemistry:

* Mineralogy: Identifying and characterizing minerals.

* Astronomy: Studying the polarization of light from celestial objects.

* Optics: Designing and characterizing optical devices.

The polarimeter's ability to unveil the subtle interactions between light and chiral molecules has
revolutionized our understanding of molecular structure and opened doors to diverse applications in
science and technology. It serves as a powerful reminder that even seemingly invisible proper

59
 REFERENCE

* Chemistry LibreTexts: Polarimetry:

https://chem.libretexts.org/Bookshelves/Organic_Chemistry/Book%3A_Organic_Chemistry_
Lab_Techniques_(Nichols)/07%3A_Polarimetry

* Khan Academy: Polarization of Light: https://www.khanacademy.org/science/physics/light-

waves/introduction-to-light-waves/v/polarization-of-light

* University of California, Davis: Polarimetry:

https://chemwiki.ucdavis.edu/Physical_Chemistry/Spectroscopy/Optical_Spectroscopy/Polari
metry

* Master Organic Chemistry: Specific Rotation:

https://www.masterorganicchemistry.com/2012/05/04/specific-rotation/

* Rudolph Research Analytical: Polarimeters:

https://www.rudolphresearch.com/products/polarimeters/

* JASCO: Polarimeters: https://jascoinc.com/products/polarimeters/

* Encyclopedia Britannica: Polarimetry in Astronomy:

https://www.britannica.com/science/polarimetry

60