Unit-2: Instrumental Methods of Analysis

Contents:

Introduction: Types of reference electrode (calomel electrode), indicator electrode (glass electrode), ion
selective electrode(solid membrane electrode).

[A] Conductometry: Introduction, conductivity cell, conductometric titrations of acid versus base with
titration curve. (Strong acid verses-Strong base). Application of conductometry.

[B] pH metry: Introduction, standardization of pH meter, pH metric titration of strong acid versus strong
base with titration curve and its application.

[C] UV-Visible Spectroscopy: Introduction, Statements of Beer’s Law and Lambert’s Law, Electronics
transitions in organic molecules, terms involved in UV-Visible Spectroscopy. Instrumentation(double beam)
and its application. Numerical: Based on absorption Laws i.e. molecular absorptivity and concentration.

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Unit-2: Instrumental Methods of Analysis
❑ Unit Objectives:
▪ On completion of this unit the students will be able to :
1. Understand the basic concepts of electrode potential generation.
2. Applications of reference electrodes.
3. Understand working of ion selective membrane.
4. Understand & apply the analytical methods in industries.

❑ Course outcome:
▪ On completion of this unit course the students will be able to Select appropriate electro-analytical
technique and method of material analysis.

❑ Text Books Used:

• T1: Engineering Chemistry by O .G. Palanna, Tata Macgraw Hill Education Pvt. Ltd.
❑ Reference Books Used
• R1: Engineering Chemistry, Wiley India Pvt. Ltd.

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Introduction:
▪ Instrumental methods of analysis involves the study of analysis of chemical solutions by using electrical
instruments to find out the concentration of unknown solution.

▪ This is the method which measures the interaction between an electrode and the solution in contact with the
analytic solution.

▪ There are a wide variety of techniques used for analysis, for examples

• 1. Gravimetric analysis (simple weighing)

• 2. Volumetric analysis (titrations)

• 3. Electro analytical methods:

• I. Potentiometry,

• II. pH metry

• III. Conductometry

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Important Applications of this Technique:

▪ Industries:

• This technique is useful for Identification of substances , to trace element detection, identification and

• characterization of final product of synthesis.

▪ Agricultural Science:

• Analysis of soil and water to find out the fertility status of the soil.

• Soil reclamation in control of soil pollution.

▪ Environmental Science:

• Applicable in atmospheric chemistry, soil chemistry, water treatment for purification, hazardous-waste

• management, industrial hygiene.

▪ In Research institutes or Laboratories.

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▪ Electrochemical cell: It is a system in which two electrodes are fitted in the same electrolyte or in two
different electrolytes which are joined by a salt bridge.
▪ Electrochemical cells are of two types.
1. Electrolytic Cell:
▪ It is a device in which chemical reaction involving oxidation & reduction is carried out by using electricity
i.e. electrical energy converts into chemical energy.
▪ Chemical reaction absorb free energy from an electrical source to drive a nonspontaneous reaction.
• Electrical E. Chemical E.
▪ For e.g. Electrolysis is used to extract many metals like Na, K, Ca, Mg, Al, etc. and manufacture of
chemicals such as NaOH, Cl2, F2, H2 etc.
2. Galvanic cell/Voltaic cell:
▪ It is a device in which a redox reaction is used to convert chemical energy into electrical energy.
▪ For example all types of batteries, fuel cells etc.
• A spontaneous reaction generates electrical energy.
• Chemical E. Electrical. E.

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REPRESENTATION OF A CELL

▪ Basic rules are used to represent electrochemical Cell.

1. The anode (-ve electrode) is written on the left side and cathode (+ve electrode) on the right hand side.
2. A vertical line or semicolon(;) indicates a contact between two phases. The anode of the cell is represented
by writing metal first & then metal ion present in the electrolyte solution. For example Zn | Zn2+
3. The molar concentration of the solution is written in brackets after the formula of ion.
For example: Zn | Zn2+ (1M)
4. The cathode of the cell is represented by writing the cation of electrolyte first & then metal with a vertical
line For Example: Cu2+ (1M) | Cu
5. Two half cells are separated by double vertical lines. Double vertical lines indicate salt bridge or any type of
porous partition between two electrolytes in galvanic cell.
6. The complete cell is represented as following. For example representation of Galvanic cell.
Zn | ZnSO4 (0.1M) || CuSO4 (0.1M) | Cu

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 Electrode Potential:
1. When a metal is placed in a solution of its ions, the metal acquires either a positive or negative charge with
respect to the solution.
2. Thus, a definite potential difference is developed between the metal and the solution. This potential
difference is called as electrode potential.
3. The potential difference is established due to the formation of electrical double layer at the interface of metal
and the solution.
4. When a metal rod is dipped in its salt solution to form a single cell, the following two changes can occur.
• a) M M+(aq) + ne- ------ Oxidation – Develops –ve charge on electrode
• b) M+(aq) + ne- M ------- Reduction -- Develops + ve charge on electrode
• 5. The magnitude of electrode potential of a metal is a measure of its relative tendency to lose or gain
electrons.
▪ Two types of Electrodes potentials:
1. Oxidation Potential: When electrode is –vely charged w. r. t. solution then it acts as anode.
2. Reduction Potential : When electrode is +vely charged w. r. t. solution then it acts as cathode.

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Development of electrode potential

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 Measurement of Electrode Potential:
1. Its not possible to measure electrode potential of the single electrode directly.
2. Only the difference in potential between two electrodes can be measured experimentally.
3. It is therefore necessary to couple the electrode with another electrode whose potential is known.
4. The electrode whose potential is known is called as reference electrode.
❑ What are the difficulties to measure electrode potential by using SHE(Standard hydrogen
electrode)?
1. Platinum used in it, gets easily poisoned by absorption of impurities from the solution and the H2 gas.
Because of absorbed impurities it is not possible to maintain stable potential.
2. It can not be used frequently in the presence of reducible ions having positive electrode potential such as
Cu, Ag, Au etc.
3. So, it is difficult to prepare and maintain stable potential by using standard hydrogen electrode.

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Reference Electrodes:
Q. What is reference electrode? Explain the construction of calomel electrode with cell
representation and labelled diagram. Give its limitations. 2019, 20, 22, 23(SEM-1 & 2)

▪ Definition:
▪ Reference electrode is defined as the electrode which has stable and reproducible
potential and completes the cell when connected with another electrode.
▪ Purpose of reference electrode:
• 1. To complete the half cell.
• 2. To provide stable potential.
• 3. To measure the electrode potential and obeys Nernst Equation
▪ Types of Reference electrode:
• 1. Primary reference Electrode: Standard Hydrogen electrode
• 2. Secondary Reference Electrode: Calomel electrode, Glass electrode-Indicator
electrode, Silver-Silver chloride electrode.
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Calomel Electrode:
Q. What is reference electrode? Draw a neat labelled diagram of calomel
electrode and give its cell representation along with explanation. Give its
disadvantages. [6 marks May 2022], 3 M Jan 2025

Fig. Calomel Electrode

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Calomel Electrode Construction:
▪ Construction:
1. It is a secondary reference electrode.
2. Calomel electrode consist of a narrow glass tube.
3. At the bottom of glass tube liquid mercury is placed above which a paste of Hg and Hg2Cl2 is placed.
4. A platinum wire dipped into liquid the mercury layer to make electrical contact with electrolyte
solution.
5. Remaining portion filled by saturated KCl solution.
6. Representation of Cell:

Pt,

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Calomel Electrode Cell Reactions:
Hg2Cl2 + 2e¯ 2Hg + 2Cl¯
▪ If calomel acts as anode, then the reaction is reverse.

▪ The potential of the calomel electrode depends up on the concentration of KCl solution is used.

▪ If the KCl solution is saturated the electrode is known as saturated calomel electrode(SCE).

▪ If KCl is 1 N, it is known as Normal calomel electrode (NCE).

▪ If KCl is 0.1 N, it is known as Decinormal calomel electrode (DCE).

▪ Generally saturated KCl is used in calomel electrode as electrolyte.

❑ Applications:

1. Calomel electrode is used to measure electrode potential of other metals and alloys of metals.

2. It can be used in combination with glass electrode in PH metric analysis to measure PH value.

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The reduction potentials of the calomel electrode depends on concentration of KCl solution are
as follows.

Concentration KCl E0 Calomel electrode Name

Saturated 0.2422 Volts Saturated Calomel
1N 0.2800 Volts Normal Calomel
0.1 N 0.3338 Volts Decinormal
Calomel.

▪ Demerits/Disadvantages of Calomel Electrode

1. It should not be used above 50°C.
2. It involves handling of poisonous Hg and Hg2Cl2
▪ Advantages:
Reproducibility of Calomel electrode is much better than Ag | AgCl.

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Glass Electrode(Indicator Electrode):

Ag wire coated with AgCl

Glass tube

Thin-walled glass
bulb

Fig: Glass Electrode

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Glass Electrode (Indicator Electrode):
Q. Give the construction of glass electrode with labelled diagram, its representation and application.
[4 marks March 2020]

▪ Definition: The glass electrode is a PH sensitive electrode whose potential depends up on the H+
ions concentration of a solution in which it is immersed.

1. The electrode of a cell in which the potential depends on the concentration of a particular ion is
called indicator electrode.

2. So, glass electrode is also called as indicator electrode.

3. For example, when a glass electrode is in contact with a solution, it acquires a potential which
depends up on H+ ion concentration of the solution.

4. So, glass electrode is sensitive for H+ ions and useful to determine PH of the solution.

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Construction of Glass Electrode:
1. The glass electrode consist of a very thin-walled glass bulb which is made from a low melting point
glass having high electrical conductivity at the bottom of electrode.
2. The glass membrane at the bottom is doped with alkali metals like Li+, Na+, K+ cations.
3. A silver wire coated with AgCl is dipped in the 0.1 M HCl solution and inserted up to the glass
bulb.
• Representation of glass electrode:
▪ The glass electrode is represented as,

Ag, AgCl 0.1 M HCl H+ (Test solution)

and potential E = E0G + 0.0591PH

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Working:

1. When glass electrode is placed in a solution, the potential develops across the
glass membrane due to the difference in concentration of internal and external
test solution.
2. Where E0G is the known potential of glass electrode.
3. The glass membrane acts as an ion exchanger i.e. exchange of Na+ of glass with
H+ of solution.
4. The potential of the glass electrode is determined using standard calomel
electrode by connecting glass electrode with calomel electrode.

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Glass Electrode - Determination of PH of the solution.
▪ Determination of PH of the solution.
1. A glass electrode is coupled with the calomel electrode to determine the pH of the solution.
2. The electrodes are dipped into the solution of unknown PH. The cell is represented as,

Pt Hg, Hg2Cl2 Cl- unknown H+ solution glass

▪ Ecell = Ecal – EG = 0.2422 – (E°G + 0.0591 pH)

pH =

▪ where E0G is the potential of glass electrode when it is in contact of known PH solution.

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Advantages of Glass electrode:

1. It is portable and compact.

2. Gives accurate and quick results.
3. Equilibrium easily obtained.
4. It is stable electrode and can be used in presence of strong oxidizing and
reducing agents.
5. It can detect & determine H+ ions in presence of other ions.
6. It is simple to use.

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Ion Selective Electrodes:
• Q. Give the composition of membrane of the ion selective electrode used for determination of
H+, F- & Cl- (3 Marks SEM-1 Oct. 2019, SEM-1 & 2, 2023)

1. Ion Selective Electrode: Definition: Ion selective electrodes are membrane electrodes which are
acting as a sensors, that converts the activity of a specific ion into an electrical potential.

2. Hence, they are called as chemical sensors.

3. OR The electrode that determines the activity of a selective ions in aqueous solution by measuring
the electrical potential are called ion selective electrodes.

4. They are also known as a specific ion selective electrodes.

5. Thus, specific ions are detected by use of a selective membrane which allows only specific ions to
sense.

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Function of Ion-selective Electrode: How it works?
▪ In this electrode a suitable membrane separates the two solutions containing same
ions of different concentrations and potential is developed on either sides of the
membrane (E1 and E2)

Solution 1 Membrane Solution 2

E1 E2

▪ Ion-selective electrodes are sensitive for a particular ion even in the presence of
the other ions in the solution.
▪ Hence, they are also known as chemical sensors.

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Types of Ion-Selective Electrodes
❑ Five types of ion – selective electrode.
▪ Based on the type of membrane used in ion-selective electrode, these can be classified into various
types as-
+
1) Glass membrane electrode – For detection of H ions
_ _
2) Solid – State membrane Electrode – For detection of F & Cl ions – Halogen ions
3) Liquid-liquid membrane electrode – For detection of Ca-deficiency in blood serum
4) Gas Sensing membrane electrode – For detection of gases like NH3, NO2, SO2, CO2, H2S
5) Enzyme based membrane / Biochemical electrode – For detection of urea.

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Solid – State membrane Electrode:
Q. What are ion selective electrodes? Give the composition of membrane of the ion selective
electrode used for determination of F- & Cl- . [March 2020] 3 Marks
• Ion Selective Electrode: Definition: Ion selective electrodes are membrane electrodes which are
acting as a sensors, that converts the activity of a specific ion into an electrical potential.

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Solid – State membrane Electrode:
I. Solid State Membrane Fluoride ion determination:
1. Composition of membrane: In this electrode solid state ionic conducting membranes are used. These
membranes are made from mixture of ionic compounds i.e. LaF3 + EuF2 mixture.
2. The Crystal of Lanthanum Tri-fluoride with europium difluoride(LaF3 + EuF2) is sealed at the bottom of
the polymeric container containing internal reference solution like NaF + NaCl or KF + KCl and consisting
of reference electrode. The F ̶ ions are detected by creating potential at the boundary surface of membrane.
3. This electrode is used for the determination of F ̶ ions in water.
II. Solid state membrane For chloride ions Detection:
1. Composition: In this pressed pellet of Ag2S + AgCl is used along with epoxy resin as a porous membrane.
_
2. This membrane is sensitive for Cl ion detection.
3. The electrode is made up of Teflon body.
4. When the electrode membrane is in contact with a solution containing chloride ions an electrode potential
develops. This potential is measured against a constant reference potential with internal solution KCl or
NaCl.

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Diagram for Solid state electrode for Chloride ion detection

Fig: 2. Solid state electrode for Chloride detection

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Conductometry:
What is conductometry?
Conductometry is an electrochemical analytical technique used to measure the electrical conductivity
of an electrolyte solution using a conductometer determines concentration of ions:
▪ Principle:
▪ The mobility of ion is affected by factors like the charge on ion, size and mass of ion and extent of
solvation.
▪ Thus, the conductivity of electrolyte solution depends upon the size of ions, concentration of ions
and nature of electrolyte solution.
▪ Conductometry is widely used in neutralization titration, precipitation titration, chemical kinetics
and plant laboratories etc.

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Important Terms & Laws Involved in Conductometry

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Basic Terms In Conductometry

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Basic Terms In Conductometry

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Basic Terms In Conductometry

6. Cell Constant

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Basic Terms In Conductometry
7. Equivalence Conductance: (λv )

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Basic Terms In Conductometry
8. Molar Conductance: (µ)

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Conductivity Cell:
Q. Explain the construction of conductivity cell with labelled diagram. (3 Marks, Oct. 2019, 20, 23

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Diagram of conductivity Cell

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Factors Affecting Conductivity:
Q. Explain any four-factor affecting the conductivity. ( 4 marks , SEM-II, 22)
1. Effect of dilution:
▪ Conductance(C) of the solution increases on dilution because it depends up on the number of movable
ions present in the solution.
▪ The specific conductance(k) decrease on dilution. On dilution results into increased in dissociation of
ions & the number of current carrying ions per ml decreases i.e. degree of ionization increases. Thus, ions
present per centimeter cube of the solution becomes less.
2. Equivalent conductance(λv ) and molar conductance (µ) both increases on dilution of the electrolyte
solution.
3. Mobility & Size of Ions: If smaller is the size of ion, greater will be its mobility and higher the
conductivity. For e.g. H+ ions are more conductive than NH4+, Na+ ions.
4. Charge on ion: Higher the charge on ion greater will be its conductivity.
5. Temperature: At higher temperature conductivity of an electrolyte increases because at high temperature
kinetic energy of ions increases.

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Conductometric Titration
▪ It is an instrumental method of analysis in which end point of the acid-base titration is determined by
measuring conductance of analyte solution using conductometer is known as conductometric
titration.
▪ There is no need to use indicator in this titration.
▪ Principle :- The conductivity of electrolyte solution depends upon the size of ions, concentration of
ions and nature of electrolyte solution.
▪ Types of Titrations:
1. Strong acid versus strong base titration(HCl VS NaOH)
2. Weak acid versus strong base. (CH3COOH VS NaOH)
3. Weak base versus strong acid.(NH4OH VS HCl)
4. Weak acid versus weak base.(CH3COOH VS NH4OH)

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1. Strong acid versus strong base titration:
Q Explain the three stages of conductometric titration between strong acid and strong base with
titration curve and reaction.

▪ Conductometric Titration Curves

1) Strong Acid with Strong Base
▪ Consider the titration of HCl with NaOH, known volume of HCl is taken beaker in which conductivity cell is
dipped and burette is filled with NaOH.
▪ Reaction HCl + NaOH NaCl + H2O

▪ Mobility of ions on electrolytic dissociation:

HCl H+ + Cl-
NaCl Na+ + Cl- Mobility of H+ > Na+ ----- Conductance decreases
Na+ and Cl- salt ions havinig minimum conductance
NaOH Na+ + OH-

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Conductometry: Strong acid versus strong base titration
❑ Explanation:
1. Before Getting End Point:
▪ HCl is strong acid and therefore dissociates completely to produce highly mobile H+ ions, hence initially the
conductance is high.
▪ During titration the highly mobile H+ ions are replaced by slow moving Na+ ions, hence conductivity goes on
decreasing until the end point.
2. At The End Point:
HCl + NaOH NaCl + H2O
▪ At complete neutralization reaction we get lowest conductance due to formation of less conductive salt NaCl
because Na+ & Cl- both ions are having least conductivity.
3. After The end Point:
▪ If further NaOH is added after end point, the conductance values will increase because of strong nature of
NaOH solution which dissociate very easily into Na+ & OH

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Conductometry: Strong acid versus strong base titration
❑ Calculation:
• If a graph of conductance verses volume of NaOH added is plotted the following ‘V’ type graph
is obtained.
• From known normality of base and volume of equivalence point we can calculate normality and
strength of acid.
• 1ml 1N NaOH = 36.5 mg of HCl.

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❑ Advantages of Conductometric Titration:
1. This method of analysis Can be used for colored and turbid solutions.
2. It Can be used for determining concentrations of weak base and weak acid or mixture of acids.
3. Does not required indicators.
4. Gives accurate result.
5. This method is useful for analysis of polybasic acid solutions.

Q. Explain any three applications of conductometry. 3 M Jan 2025

❑ Applications:
1. Chemical analysis: Used to find out concentration of ions from solution
2. Quality control: Used to monitor the quality of various products.
3. Industrial processes: for e.g. electroplating, electro-electro-refining electro-dialysis etc.
4. In conductometric titration.
5. In medical diagnostics.

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(B) pH Metric Analysis:

What is pH-metry?
pH-metry is the scientific measurement of the acidity or alkalinity (pH)
of a water-based solution, usually using an electronic pH meter to
measure hydrogen-ion activity.

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Concept of pH:

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Types of Buffer solutions:
What is a buffer solution? Give its types with example each. ( 3 Marks, Jan. 23, May, 23)

1. Acidic buffers: (CH3COOH + CH3COONa i.e. weak acid & salt of weak acid with strong base)

▪ These are prepared by mixing weak acid along with a salt of weak acid and a strong base.

▪ For example, a solution containing equvimolar quantities of CH3COOH and CH3COONa maintains its pH
value around 4.74.

2. Basic buffer: (NH4OH + NH4Cl i.e. weak base & salt of weak base with strong acid)

▪ These are prepared by using a weak base along with a salt of weak base and a strong acid.

▪ For example: Buffer solution of ammonium hydroxide i.e. NH4OH and ammonium chloride i. e. NH4Cl.

▪ A solution containing equimolar quantities of ammonium hydroxide and ammonium chloride maintains its
pH value around 9.25.

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❑ pH Metric Titration:

▪ A pH metric titration is the titration in which end point of acid-base titration is determined by
measuring pH of the solution with the help of pH meter(without use of indicator).

▪ pH metric titration can be used for acid-base titration, redox titration by using suitable electrode.

▪ For e.g. use of glass electrode.

▪ Principle of pH metric titration:

▪ In pH metric acid-base titration pH of analyte solution is measured during titration, the sharp or
sudden change in pH value indicates end point of the neutralization reaction.

▪ Before starting actual titration, first standardization/calibration of pH meter is required.

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❑ Standardization of pH meter:
Q. Write any two advantages of instrumental methods of analysis. Give stepwise process for
calibration of a pH meter. ( 3 marks, Jan. 23, May 23)
▪ Buffer solutions of PH 4, 7, 9.2 are used for standardization/calibration of PH meter.
▪ A digital PH meter is standardized/calibrated as follows-
1. Connect PH meter to electricity and connect glass electrode to PH meter.
2. Clean/rinse the glass electrode with distilled water.
3. Immerse the electrode in the solution of known PH
4. Adjust temperature knob at 20°C.
5. If the electrode is placed in buffer solution of PH= 7 then display should show the same
reading otherwise adjust the PH = 7 on the display of PH meter.
6. Similarly, PH meter is calibrated for PH 4 as well as PH 9.2 using respective buffer
solution.

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pH metric Titration: Instrumental Setup

Fig: PH metric Titration Instrument Assembly

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❑ pH metric Titration: Strong Acid (HCl) against Strong Base (NaOH):
Q. Explain various stages of pH metric titration for strong acid and strong base with titration
curve and reaction involved in it. Give any two applications of pH metry. 6 M Jan 25 (5
Nov//Dec 19, 22, 23)
▪ Procedure:
1. Pipette out known volume of HCl in beaker.
2. Immerse the indicator electrode i.e. glass electrode in the beaker solution.
3. Fill the burette with 0.1 N NaOH.
4. Measure the PH of acid HCl solution at zero addition of NaOH.
5. Add 0.5 ml of NaOH to the acid in the beaker. Stir well and note down the PH.
6. Continue titration by adding each time 0.5 ml of NaOH and note down the PH till a sudden increase in PH is
observed.
7. After getting sharp change in PH value, continue titration for 4 – 5 readings i.e. PH values remains nearly
constant.

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pH metric Titration: Strong Acid (HCl) against Strong Base (NaOH):
1. Before Getting End Point:
i. In strong acid verses strong base titration both the acid and base dissociates completely.
ii. Initially strong acid shows lowest acidic pH i.e. nearby 3 to 4 pH.
iii. When Strong base NaOH is added each time 0.5 ml, pH value starts increasing till end point,
because of H+ ions are getting neutralizing with NaOH to form neutral H2O molecules NaCl salt.
2. At The End Point:
i. At complete neutralization of HCl with NaOH, PH value suddenly increases from the neutral
range to basic i.e. from 7 to 9 pH.
ii. The reaction at the end point as,
NaOH + HCl Na+ + Cl- + H2O
3. After End point:
After complete neutralization if excess NaOH added from burette, pH increases slowly up to basic
pH = 12 due to alkaline NaOH solution.

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Titration Curve:
Titration curve is obtained by plotting PH value versus ml of base added from burette.

Fig: Titration Graph of PH metric Titration

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Differential Graph:
1. Correct end point of the reaction can be detected from the differential graph.
2. Another Graph can be plotted as ∆PH/∆V vs ml of NaOH added.
3. ∆PH is the change in PH caused by addition of the ∆V volume of burette solution.
4. The volume corresponding to peak of the curve gives the equivalence point of titration.

Fig: Differential Graph of PH metric Titration

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Applications of pH metry:

1. In Environmental monitoring: Used to measure pH levels of water.

2. In food & beverage industry: used to measure pH of various foods and beverage
products.
3. In chemical industry: Used to measure pH levels of chemicals.
4. In agriculture: Used to measure pH level of soil to check fertility of soil.
5. In medical Lab: Used to measure pH of blood, urine etc.

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❑ UV-Visible Spectroscopy:

➢ Introduction:
▪ Spectroscopy: Definition:
▪ Spectroscopy is the branch of science that deals with the study of interaction of
electromagnetic radiation with matter.
▪ It is the most important and powerful tool available for the analysis of wide range
samples or substances.
▪ Spectroscopic methods of analysis are based on measurement of the electromagnetic
radiation emitted or absorbed by the sample.
▪ Physical and chemical properties of the substance are directly related with their structure.
▪ With the help of spectroscopic technique, we can determine structure, saturation,
geometry and functional group of compound can be determined.
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Electromagnetic Radiation:
1. Electromagnetic radiation is a form of energy that is transmitted through space with high
velocity.
2. EMR consist of discrete packets of energy which are called photons.
3. A photon consists of an oscillating electric field and magnetic field.
4. The electric and magnetic fields are perpendicular to each other and perpendicular to the
direction of propagation of light.
5. EMR propagates in space without any material medium in the form of transverse waves.

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Basic Terms of EMR:

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Electromagnetic Spectrum:

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1. Lambert’s Law:
➢ Q. State and give mathematical expression of Beers and Lambert’s Law. 3M [July-23, Dec. 2019]
➢ Lambert’s Law:

▪ Statement: When a beam of monochromatic light is passed through a solution, decrease in

intensity of incident radiation is directly proportional to path length at constant
concentration.
▪ Let 𝐼o will be the intensity of incident radiation.
▪ χ be the thickness of the solution, and dI is decrease in intensity of radiation then lambert’s Law
mathematically written as,

▪ In other word absorbance (A) is directly proportional to path length of solution x at constant concentration.
Aα𝑥

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Fig: Lambert’s-Beer’s Law

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 2. Beer’s Law:
2. Beer’s Law:
▪ Statement: When a beam of monochromatic light is passed through a solution, decrease in
intensity of incident radiation is directly proportional to concentration at constant path
length.
▪ Let I0 will be the intensity of incident radiation
▪ Let will C be the concentration of solution, dI is the decrease in intensity of radiation.
▪ Then, Beer’s Law mathematically can be written as,

▪ Or Absorbance (A) is directly proportional to the concentration (C)of solution at constant path length.
AαC

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Fig: Beer’s Law to find out concentration of Unknown solution

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❑Unsolved problems on Lambert-Beer’s Law:
• 1. Calculate molar absorptivity of 0.5 × 10 -3 M solution which has an absorbance
of 0.17, when pathlength is 1.3 cm
[Ans: ε 261.54 dm3 / mol / cm]
2. What is the concentration of coloured complex if it show an absorbance of 0.25
in glass cuvette with optical path of 10 mm and complex has molar absorptivity
5890? [Ans: 4.21 × 10 -5 M]

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UV-Visible Spectroscopy:
▪ The visible radiation region extends from 400 nm – 800 nm and the UV radiation region extends from
10 nm – 400 nm.

The UV region is subdivided into two spectra regions as,

1. Near UV region: from 200 nm – 400 nm.

2. Far or vacuum UV region: below 200 nm.

▪ Far UV spectrum is studied under vacuum condition.

▪ The common solvent used for preparing sample to be analyzed is either ethyl alcohol or hexane.

❑ Photon Energy of UV-Visible region:

▪ The Visible region Photon Energy: 36 – 72 kcal/mole

▪ UV region Photon energy:143 kcal/mole.

▪ These energies are sufficient to excite a molecular electron to higher energy level.

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Absorption of UV-Visible radiation – Electronic Transition:

▪ Principle UV-Visible Spectroscopy: When energy absorbed from UV-Visible region by a

molecule, it causes excitation of valence electrons from lower energy to higher energy i.e. from
bonding molecular orbital, non-bonding orbital to anti-bonding molecular orbitals.

➢ Three types of electrons are involved in organic molecules.

1. Sigma (𝛔) Electrons: These electrons are involved sigma bond formation in saturated
hydrocarbons like C-H bond or C-C bond.
2. Pi (𝜋) Electrons: These electrons involved in pi bond formation in unsaturated compounds in
which double or triple bonds are present. Pi bonded electrons may be present along with functional
groups like -C=O, -CHO, -COOH etc.
3. Non-bonding – n-Electrons: These are non-bonding or lone pair of electrons and are not involved
in bond formation between atoms in molecules. For e.g. Lone pair of electrons present on oxygen,
nitrogen etc.

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Types of Electronic Transitions:

Explain with diagram the possible electronic transition occur in organic molecules on absorption of UV-
radiations. Also state forbidden electronic transition. 6M July-23, March-23, August-22, Dec.-2019
When a molecule absorbs energy in UV or Visible region, its electrons are promoted from a bonding, non-
bonding molecular orbitals to an anti-bonding molecular orbitals.
There are four main electronic transitions.
  →  −Transition
2.  →  Transitions
 n →  Transitions
4. n →  Transitions
▪ The energy required for various electronic transitions show the following order.
▪  →    →   n →   n → 
▪  →  and  →  are considered as forbidden transitions. Practically not possible.

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Electronic Transitions in organic compounds:

Fig: Different Types of Electronic Transitions

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Explanation of Electronic Transitions with Examples:

1. 𝛔 -  Transition :
1. A sigma (𝛔) – electron from bonding orbital is excited to the corresponding antibonding sigma starred (𝛔*)
orbital.
2. The energy required is large for this transition.
3. For example, methane (which has only C-H bonds, and can only undergo  →  transitions shows an
absorbance maximum at 125 nm.
4. Thus, saturated hydrocarbons absorb very short wavelength below 180 nm.
  →  starred Transition :
1. Pi () – electron in a bonding orbital is excited to corresponding anti-bonding Pi () orbital.
2. Compounds containing multiple bonds like alkenes, alkynes, nitriles, aromatic compounds undergo  → 
transitions.
3. Molecules containing two or more conjugated double bonds absorb visible radiation above 200 nm and can be
easily recorded.
4. Alkene generally absorb in the region 170-205 nm. It is also called ethylenic E-band.
5. Conjugated compounds show absorption in the range 210 – 280 nm.
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❑3. n - 𝛔* Transition:
1. Saturated compounds containing functional group with lone pair of electrones (non-bonding
electrons) are capable of n - 𝛔* transitions. e.g. Alcohol(-OH), amines(-NH2), halides(-X),
ethers(R-O-R) etc.
2. These transitions usually need less energy than  → * transitions.
3. They can be initiated by light whose wavelength is in the range 150 nm - 250 nm.
4. The number of organic functional groups with n →  peaks in the UV region is small.
❑4. n - 𝜋*): Transition:
1. Compounds containing double bond involving heteroatoms like C=O, nitrile N=O
undergo such transition.
2. n →  Transitions require minimum energy and show absorption at longer wavelength
around 300 nm.
3. The aldehydes and ketones having C=C bond adjacent to the carbonyl group show
transition in the range 300 – 350 nm.
4. Thus n →  and  →  electronic transitions show absorption in region above 200 nm
which is accessible with UV-Visible spectrophotometer.

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Terms Involved In UV-Visible Spectroscopy:
➢ Q. Define the following: 1Mark each [Dec. 2019, August-22, March-23, ]
1. Chromophore
2. Auxochrome
3. Bathochromic shift (Red Shift)
4. Hypsochromic Shift (Blue Shift)
5. Hyperchromic Effect
6. Hypochromic Effect

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1. Chromophore
Definition: Chromophore is a functional group containing multiple bonds
capable of absorbing radiations above 200 nm due to  →  transitions and n →
 transitions
Or another definition is A chromophore is the part of a molecule responsible for its colour
appearance.
The colour arises when a molecule absorbs certain wavelengths of visible light and
transmits a coloured light.
1. For e.g. β-carotene in carrot.
2. Compounds containing functional groups like NO2, N=O, C=O, C=C, C=S etc.
3. Non conjugated carbonyl group of compound gives weak absorption band in 200 to 300
nm region. For e.g. Acetone λ max = 279 nm, cyclohexanone λmax = 291 nm.
4. Double bonds in conjugation, λ max shifted to longer wavelength.
• For e.g. 1, 5 hexadiene λmax = 178nm ; CH2=CH-CH2-CH2-CH=CH2
• 2, 4 hexadiene λmax = 227 nm , CH 3-CH=CH-CH=CH-CH
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 Examples of Chromophores: Natural Fruits:
• β- Carotene λmax = 505 nm

β- Carotene λmax = 505

nm
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 Examples of Chromophores: Natural Fruits:
▪ Lycopene Pigment: It is a natural pigment Lycopene imparts red color to the
tomato, watermelon, straw berry etc. Lycopene shows visible light radiation at
about 445 nm – 503 nm.
▪ Chlorophyll is also a natural pigment present in plants leaves.

Fig : Lycopene

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❑ 2. Auxochrome:
▪ Definition: An Auxochrome is a saturated functional group with lone pair of electrons (non-bonded) that
does not absorb radiation in near UV region but when attached to a chromophore alters the wavelength
and intensity of absorption.
▪ It causes shift in absorption to longer wavelength.
▪ It is a group which extends the conjugation of a chromophore by sharing of non-bonding electrons.

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❑ 3. Bathochromic shift (Red Shift):
▪ Definition: When absorption maxima (λmax ) of a compound shift to longer wavelength it is
known as bathochromic shift or red shift.

▪ This effect is due to the presence of an auxochrome or by the change of solvent.

▪ For e.g. An auxochrome group like –OH, -OCH3 causes absorption of compound at longer
wavelength.

▪ In alkaline medium, p-nitrophenol shows red shift. Because negatively charged oxygen delocalizes
more effectively than the unshared pair of electron i.e. non- bonding electron on –OH.

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❑4. Hypsochromic Shift (Blue Shift):
▪ Definition: When absorption maxima (λmax ) of a compound shifts to
shorter wavelength, it is known as hypsochromic shift or blue shift.
▪ This shift is caused due to 1) Removal of conjugation or 2) Changing polarity of
solvent.
▪ For e.g. Aniline shows blue shift in acidic medium, it loses conjugation.

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➢ 5. Hyperchromic Effect:
▪ Definition: When absorption intensity (ε) of a compound is increased, the effect is known as
hyperchromic effect.
▪ If auxochrome introduces to the compound, the intensity of absorption increases.
▪ For example: in 2-methyl pyridine, methyl group increases conjugation.

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❑ 6. Hypochromic Effect:

▪ Definition: When absorption intensity (ε) of a compound is decreased,

it is known as hypochromic shift.
▪ It occurs due to introduction of groups that disturbs the original geometry of
the molecule.

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❑ Shifts on absorption and Intensity of the compound can be summarized as,

Fig: Absorption and Intensity shift

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❑Instrumentation of Double Beam UV-visible Spectrophotometer:

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❑ Instrumentation of Double Beam UV-visible Spectrophotometer:

▪ Various components of UV-visible Spectrophotometer and their functions are as

follows:
1. Light Source
2. Monochromator
3. Sample Holder/Cuvette Tube
4. Detectors
5. Amplifier and Recorder

1. Light Source (Sources of Radiations):

▪ Function: Following source material provide the entire UV-Visible radiation band:
a) Deuterium lamp – covers the UV – 190 nm - 350 nm
b) Tungsten lamp – covers 330 nm -780 nm (Visible range)
c) Xenon arc lamp- 200 – 1000 nm.

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2. Monochromator:
▪ Monochromators are used in this spectrophotometer For e.g. Quartz, prism are used as monochromator,
Glass prisms are used for visible region, quartz prisms are used for UV region.
▪ Function: Monochromator converts polychromatic radiation beam of light into monochromatic light
with a particular wavelength.
▪ By rotating monochromator, a particular wavelength is made to fall on the sample cell.
3. Sample Holder/Cuvette Tube:
▪ Function: Sample holders or cuvette tubes are used to hold sample or reference solution.
▪ There is a special glass tube called cuvette which does not absorb the light.
▪ Quartz cuvette is used for UV region and glass cuvette for visible region.

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4. Detector:
▪ Function: Transmitted light from sample holder falls on the detector where it is converted into
electric current.
▪ It converts light energy directly into electrical energy.
▪ The output current of photodetector is directly proportional to the intensity of light falling on it.
▪ For Ex. Photovoltaic cell, photomultiplier tube, phototubes are used as detector.
5. Amplifiers and Recorders:
▪ Function: Signal received from detector is amplified and read on recorder.
▪ Recorder can record output either in spectral graph or in the form of digital values as
absorbance or transmittance..

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❑Applications of UV-Visible Spectroscopy:
➢ Q. Explain any five applications of UV-Visible spectroscopy. 5 M [Dec. 2019, July-23]
1. Qualitative Analysis –
i. Detection of functional group: This technique is used to determine presence or absence of
chromophore.
ii. Extent of conjugation: Extent of conjugation can be detected by UV-Visible spectroscopy.
iii. Double bond conjugation: Determination of conjugated and non-conjugated double bonds
iv. Geometric Structure: Detection of cis and trans isomers of organic compounds.
v. Detection of impurities
vi. To study Chemical Kinetics:
▪ As Absorbance is directly proportional to concentration of reactant or product, with the progress of
reaction the change in extent of absorption per minute can help to study the kinetics of the
chemical reactions.
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2. Quantitative Analysis:
▪ It can be used to find out molar concentration of the solution by Beer’s Law verification.
▪ Determination of molecular weight.

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 Question Bank
Questions: 5 / 4 Marks

1. Give the principle and construction with diagram of glass electrode. Explain how PH of a solution can be
determined using glass electrode.
2. What is reference electrode? Explain the construction with diagram of calomel electrode and Glass
electrode.
3. What is reference electrode? Explain the construction with figure of calomel electrode and glass electrode.
4. Give the principle and construction with diagram of glass electrode. Explain advantages of glass electrode
and how pH of a solution can be determined by using it.
5. Define ion selective electrode. Why it is called as chemical sensors? Explain the composition and working
of any one of ion selective electrode with diagram.
6. Give the composition and diagram of gas sensing membrane electrode. Discuss the procedure for its use in
determining ammonia in solution.

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7. Define ion selective electrode. What are important properties of ion-selective electrode membrane?
Explain the composition and working of any one of ion selective electrode.
8. Give the composition and working of enzyme-based electrode. How it can be used for estimation of urea.
9. Explain the various stages of the titration curve in the conductometric titration of strong acid Vs strong
base.
10. Draw and explain the titration curve in the conductometric titration of strong acid Vs weak base.
11. Give the reaction and discuss the conductometric titration curve of weak base (in burette) against strong
acid.

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