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Lesson-1

Analytical Chemistry: Principles, Applications and Types

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Course Name Soil and Water Chemistry

Lesson 1 Analytical Chemistry: Principles, Applications and Types

Course Revisor Name Dr. Rajdeep Dutta
University/College Name Assam Agricultural University, Jorhat
Course Reviewer Name T V Ramana
University/College Name Sri Venkateswara Veterinary University,Tirupati

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Lesson 1

Objectives:
1. To understand the principles of different analytical chemistry techniques
2. To understand different analytical chemistry concepts

Glossary of terms:
Volumetry: Volumetry is measurement of volume of a solution of known
concentration which is used to determine the concentration of the analyte.
Gravimetry: It is a technique through which the amount of an analyte (the
ion being analysed) can be determined through the measurement of mass.
Gravimetric analyses depend on comparing the masses of two compounds
containing the analyte. The principle behind gravimetric analysis is that the
mass of an ion in a pure compound can be determined and then used to
find the mass percent of the same ion in a known quantity of an impure
compound.
Standard solution: Standard solution is the one whose concentration or
strength is accurately known. It is also sometimes referred to as “titrant”.
Titration: The Process of adding a standard solution from a volumetric
burette to a solution of the unknown concentration in the conical flask until
the reaction is just complete is known as “Titration”.
Indicator: The end point or equivalent point of titration is recognized with
the help of a chemical reagent called “Indicator”.

Normality: It is a measure of concentration equal to the gram equivalent

weight per litre of solution. Gram equivalent weight is the measure of the
reactive capacity of a molecule. The solute's role in the reaction
determines the solution's normality. Normality is also known as the
equivalent concentration of a solution.

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Molarity: It is a unit of concentration measuring the numbers of moles of

a solute per litre of solution. Molarity is denoted with 'M'. A 1 M solution
contains 1 mole of solute per litre of solution.

Molality: It is the number of moles of solute per kilogram of solvent.

Solutions expressed in molal concentration are denoted with 'm'. 1 m
solution contains 1 mole of solute per kilogram of solvent.
Formality: It is defined as the number of formula mass of any solute
dissolved in one litre of solution. A solution is said to be one formal (1 F) if
it contains one formula mass of the substance in one litre of solution.
Standard curve: A standard curve also known as Calibration curve, is a type
of graph used as a quantitative research technique. Multiple samples with
known concentration are measured and plotted on the graph, which then
allows similar concentrations to be determined for unknown samples by
interpolation on the graph. The samples with known concentration are the
standards, and the graph is the standard graph.
Nomograph: These are graphical representations which are used by simply
applying a straight edge across the plot through the points on scales
representing independent variables, which then crosses the corresponding
datum point for the dependent variable; the choice among independent
and dependent variable is arbitrary so that each variable may be
determined in terms of the others.

E-lecture:
1. What is Analytical Chemistry: Analytical chemistry is application of
chemical knowledge for characterising the composition of matter, both
qualitatively and quantitatively (the identification of matter under study is
performed using qualitative analysis while quantitative analysis is used to
determine how much (relative concentration or total amount) of the
substance is present in the analyte . Methods used in analytical chemistry

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are based on chemical and physical principles to study a substance which

is to be analysed.
1.1 Classical methods of analytical chemistry:
A) Classical Qualitative methods: These methods can further be divided
into
(a) Separation methods &
(b) Identification methods
(a) Separation methods:
i) Precipitation: This is a gravimetric method in which the reactants and
products of a chemical reaction are used to analyse a substance. The
metallic ions of many elements may enter into a reaction with the negative
ions to form a new insoluble substance called the precipitate, which settles
to the bottom of the solution. This is filtered out and washed.
ii) Extraction: This is a procedure in which a substance is removed from a
matrix, both being in two immiscible phases. The most common technique
for extraction of a compound from an aqueous solution is liquid-liquid
extraction (LLE). Solid phase extraction (SPE) is a widely used method of
sample preparation used to separate and enrich purified components from
aqueous solution.
iii) Distillation: This is process of evaporation and condensation by which
a component is separated out of a mixture of liquids. It is based upon the
difference in boiling points or volatility between the various substances
present in the mixture.
(b) Identification methods: These methods are based on colour, odour,
melting and boiling points, radioactive or reactive properties of the
analyte.
B) Classical Quantitative methods:
(a) Titration method: It includes acid-base or complexometric titration.
Acid-base titration helps in quantifying an unknown acid or base in solution
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by finding its concentration based on the exact amount of corresponding

base or acid required to neutralize it. Complexometric titration is a
volumetric analysis where end point of titration is the formation of a
characteristic coloured complex, which is usually a metal complex.
(b) Gravimetric method: It uses precipitation or volatilization to determine
the mass of the analyte from its ionic mass. Precipitation, Volatilization or
electroanalytical methods are mostly used.
(c) Coulometric method: These methods help to determine the mass of
matter transformed by electrolysis from the amount of electric current
used or produced during the reaction.
1.2 Instrumental methods: Instrumental methods are mostly used in
advance level of research and the same instrument may be used to
separate, identify and quantify the analyte.
A) Instrumental separation methods:
(a) Chromatography: This method uses a mobile phase into which the
mixture is introduced, which then passed through the stationary phase.
Due to the different rates at which the components of the mixture travel
through the stationary phase, they can be separated.
There are different types of chromatography:
(i) Paper or thin layer chromatography
(ii) Gas chromatography
(iii) Ion-exchange and size-exclusion chromatography
(iv) Reversed-phased chromatography, Two-dimensional
chromatography, Hydrophobic interaction chromatography

Special Note
High Performance Liquid Chromatography (HPLC) is an advance technique in
which the analyte is contained in a mobile phase passed through a stainless-steel column
which is between 1 and 25 cm long and less than 1.0 mm to 4.6 mm internal diameter,
filled tightly with micro size particles allowing the various components of a complex
non-volatile mixture to separate rapidly. The final concentration is determined using
computations based on the outputs at varying amounts of organic compound. 5
 Soil and Water Chemistry

(b) Electrophoresis: In this method, an electric field is used to separate

particles in a fluid because of development of a charged interface between
them.
(c) Field Flow Fractionation: In this technique, a fluid suspension or
solution is fed through a narrow channel and a field is applied right angles
to it, causing the particles within the field to separate because of their
varying mobility in response to the field force.
B) Instrumental qualitative and quantitative methods: These methods
may utilize light, electromagnetic, heat and combinations of these to
identify the analytes.
(a) Spectrometry: This method can isolate particles based on various
properties like mass, momentum or energy. It includes techniques such as
ion-mobility spectrometry based on mobility of ionised particles in a carrier
gas, or mass spectrometry which measures the ratio of mass to charge of
ions.
(b) Spectrophotometry: This method is based on Beer-Lambert law. Here
a material or component is identified by its reflection or transmission of
light, which depends on the wavelength. Visible, ultraviolet or near-
infrared light may be used in this method. This method is widely used to
study gases, solids or solutions.

Beer-Lambert Law
This law states that the absorbance of a light absorbing material is proportional to its
concentration in solution. The general Beer-Lambert Law is usually written as
A = elc
Where
A is the measured absorbance

e is the extinction coefficient of the substance- unique for each substance

l is the sample path length measured in centimetres (i.e. the width of the cuvette
almost always 1 cm)
c is the molar concentration of the solution (expressed in terms of molarity)

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(c) Potentiometry: It is a method of analysis in which the concentration of

an ion or substance is determined by measuring the potential developed
when a sensitive electrode is immersed in the solution of the species to be
determined. Commonly available pH meters are designed based on this
principle.
(d) Conductometry: Conductometry is the measurement of the electrical
conductivity of a solution. The main principle involved in this method is
that the movement of the ions creates the electrical conductivity. The
movement of the ions is mainly depended on the concentration of the ions.
Electrical conductivity and Total Dissolved Solids (TDS) of water samples
generally determined by instruments designed based on this principle.
(e) Turbidimetry/ Nephelometry: The principle of nephelometry and
turbidimetry is based on the on the well-known law of Tyndall Effect.
When light is passed through the suspension, part of incident radiant
energy is dissipated by absorption, reflection, and refraction while
remainder is transmitted. In fact the measurement of the intensity of
transmitted light is a function of the concentration of dispersed phase and
this becomes the basis of turbidimetric analysis.

Fig.1. Schematic Diagram of a Nephelometer

(f) Radiometry: Radiometry is the science of measuring light in any portion
of the electromagnetic spectrum. In practice, the term is usually limited to
the measurement of infrared, visible, and ultraviolet light using optical
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instruments. Irradiance is the intensity of light and is measured in watts

per square meter.
1.3 Volumetry & Gravimetry:
A) Volumetry: It is a widely used quantitative analytical method. It involves
measurement of volume of a solution of known concentration which is
used to find out the concentration of the analyte.
Some of the widely used volumetric analysis are:
a) Acidimetry: The term acidimetry refers to that part of volumetric
analysis whereby an acid solution at known concentration, along with a
specific indicator, is used to titrate a base solution and thus work out its
concentration.
b) Alkalimetry: The term alkalimetry refers to that part of volumetric
chemical analysis which enables us to work out the concentration of an
acid solution using an alkaline solution at a known concentration and a
suitable indicator.
c) Argentometry: The determination of silver or halides by the
precipitation of silver salts is known as argentometric titrations. The
sample solution is titrated against a solution of silver nitrate of known
concentration. It can be used for measurement of salinity.
B) Gravimetry: Gravimetric analysis is a technique through which the
amount of an analyte (the ion being analysed) can be determined through
the measurement of mass. Gravimetric analyses depend on comparing the
masses of two compounds containing the analyte.
Principle: Mass of an ion in a pure compound can be determined and then
used to find the mass percent of the same ion in a known quantity of an
impure compound.
For an accurate gravimetric analysis, the following 3 conditions must be
met:
i) The ion being analysed must be completely precipitated.

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ii). The precipitate must be pure compound.

iii) The precipitate must be easily filtered.
1.4 Standard Solutions: A solution whose strength or concentration is
accurately known is referred to as Standard solution.
i) The most accurate and convenient way of preparing a standard solution
is to weigh the reagent, dissolve it, and dilute the solution to a definite
volume in a volumetric flask. However, this method can only be used if the
reagent is a primary standard.
ii) In order for a reagent to be a primary standard, it must be obtainable in
pure form (generally at least 99.98% pure), stable both is pure form and in
solution, easy to dry and keep dry, and soluble in a suitable solvent.
iii) But many useful reagents do not meet those requirements, so the
reagent is dissolved and diluted approximately to the concentration
desired. The solution is then standardised by titrating it against a primary
solution. This standardised solution is called a secondary standard.
Requirements of a primary standard substance:
➢ It must be 100% pure and dry.
➢ It must be 100% soluble in suitable solvent.
➢ It should not be hygroscopic.
➢ It should get oxidized by oxygen or affected by CO2
➢ The reactions should be stoichiometric and instantaneous.
1.5 Titration & Indicators: Titration is a widely used quantitative analytical
technique. The steps involved in titration are as follows:
i) Prepare a solution from an accurately weighed sample to +/- 0.0001 g of
the material to be analysed. It is also known as Analyte.
ii) Choose a substance that will react rapidly and completely with the
analyte and prepare a standard solution of this substance. The
concentration of the standard solution should be +/-0.0001 M. It is also
known as Titrant.

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iii) Place the standard solution in a burette and add it slowly to the solution
of unknown concentration. This process is called titration. Continue the
titration until the reaction is complete; that is, until the amount of reactant
added is exactly the amount required to react with all the constituent being
analysed. This point is called the equivalence point, and can be detected
by adding an indicator to the unknown solution before beginning the
titration.
iv) An indicator is a substance that gives a visible sign, usually by a colour
change, of the presence or absence of a threshold concentration of a
chemical species, such as an acid or an alkali in a solution. The point at
which the colour change occurs is the end point of titration. An example of
indicator is methyl yellow, which imparts a yellow colour to an alkaline
solution. If acid is slowly added, the solution remains yellow until all the
alkali has been neutralized, whereupon the colour suddenly changes to
red. Indicators are classified as acid-base, oxidation-reduction, or specific-
substance indicators, every indicator in each class having a characteristic
transition range. Some of the commonly used acid-base indicators can be
seen in Fig.2.

Fig.2. Some common pH indicators (Image courtesy: Compoundchem)

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v) Measure the exact volume of standard solution required from burette

readings before and after the titration. Since the molarity of the standard
solution is known, the number of moles of titrant can be calculated. From
a knowledge of the equation for the reaction, the number of moles of
constituent present in sample can also be calculated.
1.6 Dilute solutions:
a) Dilution is the process of adding additional solvent to a solution to
reduce its concentration. This process keeps the amount of solute
constant, but increases the total amount of solution, thereby decreasing
its final concentration. Dilution can be done by mixing a solution of higher
concentration with an identical solution of lesser concentration. Diluting
solutions is a routine process in the laboratory, as stock solutions are often
purchased and stored in very concentrated forms. For the solutions to be
usable in the lab (for a titration, for instance), they must be accurately
diluted to a known, lesser concentration.
b) The volume of solvent needed to prepare the desired concentration of
a new, diluted solution can be calculated mathematically. The relationship
is as follows:
M1V1 = M2V2
Where
M1 denotes the concentration of the original solution, and V1 denotes the
volume of the original solution; M2 represents the concentration of the
diluted solution, and V2 represents the final volume of the diluted solution.
When calculating dilution factors, it is important that the units for both
volume and concentration are the same for both sides of the equation.
Example:
• 200 ml of a 1.6 M aqueous solution of NaCl is diluted with water
to a final volume of 1.0 L. What is the final concentration of the
diluted solution?

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• M1V1 = M2V2
• (1.6 M) (200 ml) = M2(1000 ml)
• M2 = 0.32 M

c) Dilutions can sometimes be visually observed.

Fig.3. Intense red colour slowly fades as the solutions become more
diluted
(Image courtesy: Lumen: Introduction to Chemistry)

c) Serial dilutions:
Serial dilutions involve diluting a stock or standard solution multiple times
in a row. Typically, the dilution factor remains constant for each dilution,
resulting in an exponential decrease in concentration. As for example, a
ten-fold serial dilution could result in the following concentrations: 1 M,
0.1 M, 0.01 M, 0.001 M, and so on. As seen in this example, the
concentration is reduced by a factor of ten in each step.

d) Serial dilutions are used to accurately create extremely diluted solutions,

as well as solutions for experiments that require a concentration curve
with an exponential or logarithmic scale. Serial dilutions are widely used in
experimental sciences, including biochemistry, pharmacology,
microbiology, and physics.

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1.7 Units of concentration:

a) Most commonly, a solution’s concentration is expressed in terms of

normality, molarity, molality, formality etc.

b) Normal solution: A normal solution contains one gram equivalent mass

of the substance in one liter of solution.

Mass of the substance in one liter

Normality = ------------------------------------------------
Gram equivalent mass of the substance

Example: 1.0 N HCl solution contains 36.45 gm of HCl in one litre of d/w

c) Molarity: It is a unit of concentration measuring the numbers of moles

of a solute per litre of solution. Molarity is denoted with 'M'. A 1 M solution
contains 1 mole of solute per litre of solution.

Example: 1.0 M HCl consists of 36.5gm HCl in 1000ml of distilled water

d) Molality: It is the number of moles of solute per kilogram of solvent.

Solutions expressed in molal concentration are denoted with 'm'. 1 m
solution contains 1 mole of solute per kilogram of solvent.

Example: 1.0 m H2SO4 solution contains 98gm of H2SO4 in 1000ml of water

e) Formality: It is defined as the number of formula mass of any solute

dissolved in one litre of solution. A solution is said to be one formal (1 F) if
it contains one formula mass of the substance in one litre of solution.
Example: 1.0 F solution of Oxalic acid (H2C2O4, 2H2O) contains 126gm.
Oxalic acid in one liter of solutions

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1.8 Standard Curve: A standard curve also known as Calibration curve, is a

type of graph used as a quantitative research technique. Standard curves
represent the relationship between two quantities. Multiple samples with
known concentration are measured and plotted on the graph, which then
allows similar concentrations to be determined for unknown samples by
interpolation on the graph. The samples with known concentration are the
standards, and the graph is the standard graph.
Example: A standard curve for protein concentration determination.

Fig.4. Standard curve example showing protein concentration

determination
To calculate the sample concentration based on the standard curve, first
we have to find out the concentration (on X-axis) for each sample
absorbance (Optical density on Y -axis) using a spectrophotometer on the
standard curve; then we have to multiply the concentration by the dilution
factor for each sample.
1.9 Nomograph: Nomographs (or nomograms or alignment charts) are
graphical representations of mathematical relationships which are used by
simply applying a straight edge across the plot through the points on scales
representing independent variables, which then crosses the corresponding
datum point for the dependent variable; the choice among independent
and dependent variable is arbitrary so that each variable may be
determined in terms of the others. Common example of nomograms are:

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use to compute the lift available for a hot-air balloon, the boiling points of
solvents under reduced pressure and the relative forces in a centrifuge.
Nomograms provide insight into mathematical relationships, are useful for
rapid and repeated application, even in the absence of calculational
facilities, and can reliably be used in the field.

Fig.5. A Pressure-Temperature nomograph

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References:
Glasser, L & Doerfler, R (2018). A brief introduction to nomography:
graphical representation of mathematical relationships. International
Journal of Mathematical Education In Science And Technology,
https://doi.org/10.1080/0020739X.2018.1527406

Lumen: Introduction to Chemistry.

https://courses.lumenlearning.com/introchem/
Compoundchem: https://www.compoundchem.com/

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