A TEXT BOOK OF

PHARMACEUTICAL
ANALYSIS

Dr .GODAY SWAPNA

SUGRIVA. DIVYA

CHINNI. SOWJANYA
 DEDICATION

THIS BOOK IS DEDICATED TO MY

BEVELOVED PARENTS AND
FAMILYMEMBERS AND MY DEAR
STUDENTS WHO HELPED ME TO PUBLISH
THE BOOK
 Contents
Foreword.................................................................................................................................................................. ii
Preface..................................................................................................................................................................... ii
Acknowledgments....................................................................................................................................................ii
Prologue/Introduction..............................................................................................................................................ii
Chapter 1 - Role of Analytical Chemistry……………………………………………………………………………………………………….15-20

Chapter 2 - Classification of Analytical methods and Advantages of instrumental methods………………………….21-30

Chapter 3 - Solution reactions……………………………………………………………………………………………………………………….31-35

Chapter 4 - Laboratory operations and Good Laboratory Procedures……………………………………………………………36-41

Chapter 5 - Steps involved in Qunatitative analysis……………………………………………………………………………………….42-46

Chapeter 6 - Clibration Glassware and instruments………………………………………………………………………………………47-55

Chapter 7 - Stiochiometric Calculations…………………………………………………………………………………………………………56-64

Chapter 8 - Titrimetric method of analysis…………………………………………………………………………………………………….65-72

Chapter 9 - Acids, bases and their salts complexex……………………………………………………………………………………….73-82

Chapter 10 - Gravimetric method of analysis……………………………………………………………………………………………….83-88

 Preface

The pharmaceutical industry is under intense pressure to increase productivity and release
new drugs into the market. Analytical chemists are challenged to find faster ways of
delivering quality data across a range of project needs. A number of approaches are being
employed to increase separation throughout. Volumetric analysis is used in high school
and college chemistry labs to determine concentrations of unknown substances. The
titrant (the known solution) is added to a known quantity of analyte (unknown solution)
and a reaction takes place. Knowing the volume of the titrant allows the student to
determine the concentration of the unknown substance. Medical labs and hospitals use
automated titration equipment for basically the same purpose.

Author name: Dr Goday Swapna

Date: 04-08-2023
 Acknowledgments
It gives me immense pleasure and contentment to acknowledge and thank all those who, in big ways and
small, have contributed for this effort.
It’s a fact that every mission needs a spirit of hard work and dedication but it needs to be put on the right
path to meet its destination and here the credits goes to the secretary and correspondent Rev Sister
G.Nirmala Jyothi , and Dr.SK.AbdulRahaman,Principal&professor,
Nirmala college of Pharmacy, Atmakuru, mangalagiri with a deep sense of gratitude for his continuous
encouragement, timely advice, cooperation, kind suggestions and providing the best facilities during this
work.
I am very grateful to my parents for their love, encouragement and support throughout my research
work. A special note of thanks to all my family members for their wishes and prayers in all my
endeavours.

I would also like to thank my friends and well wishers for their constant support and encouragement . I
thank the Almighty for showering his blessings on me to pursue and complete my research work.
 1. Role of Analytical Chemistry

1. Introduction

Analytical chemistry is as old as chemistry. Analytical chemistry acts as a foundation for other
branches of chemistry. In fact the science of chemistry came into existence as a result of human
inquisitiveness to understand the nature of an extraordinary variety of matter that surrounded him,
and this knowledge was obtained by analysing different types of material. A complex material on
analysis can give rise to a number of new and simpler constituents. Each of the constituents can be
further analysed and if this process is continued, a stage will come when a constituent will not give
anything new; such a constituent is called an element. Thus the concept of an element as a substance
which cannot be broken down into something simpler by ordinary chemical methods arose from
analytical data on different substances. The construction of chemical balance provided a quantitative
aspect to chemical analysis. With this development the study of analytical chemistry stimulated
quantitative approach to various problems of chemistry. Chemical reactions were studied on the
basis of qualitative and also quantitative changes that occurred; this resulted in the discovery of
several new compounds and the five laws of chemical combination. In an attempt to theoretically
explain these laws, Dalton developed the atomic theory which played a tremendous role in
developing chemistry into a quantitative experience and was mainly preparative in nature centred
around empirical synthetic methods. The induction of analytical approach brought about a
revolutionary transformation from magic and alchemy to quantitative scientific chemistry.

Steps in analytical procedure

Structural information about a complex compound can be obtained either by preparing it from some
simple constituents or by breaking it chemically into smaller and simpler units and then identifying
them. The former approach involves synthesis of the compound while the latter is termed chemical
analysis.

Inorganic and Organic Analysis: Chemical analysis is broadly classified as inorganic or organic
depending upon the nature of the material under examination. Elemental analysis deals with the
detection and determination of various elements present in a compound. Functional group analysis
involves the determination of certain groupings of atoms such as carboxyl group (-COOH) or
hydroxyl group (-OH) in an organic material.

 Major, Minute and Trace Constituents: A major constituent is one whose amount is 1 per cent or
more of the sample material. A minor constituent present in quantities smaller than 0.01 percent is
called a trace constituent.

 Complete and Partial Analysis: Chemical analysis is said to be complete when it involves the
determination of all the components detected qualitatively in the sample. The analysis is partial
when it aims at determining only one or a few of the components of the sample such as
determination of copper in a copper ore.

 Classical Chemical Analysis: Quantitative chemical analysis started with the application of
techniques of gravimetric procedures. In a gravimetric measurement, a known volume of sample
solution is treated with an excess of a suitable reagent which quantitatively precipitates the desired
constituent present in the sample solution. The precipitate which is of known concentration is
filtered, washed, dried and weighed. Knowing the weight of the precipitate, the amount of the
desired constituent in the test solution is calculated. For example, an excess of dilute sulphuric acid
is added to a given solution containing barium ions. The precipitated of barium sulphate formed is
filtered, washed, dried and weighed. From the weight of barium sulphate the quantity of barium in
the given solution is calculated. Because such determinations are based on the measurement of
weight, these are referred to as gravimetric determinations.

In electro-gravimetric analysis, the constituent to be determined is deposited on the electrode by

passing electric current through a suitable electrolytic cell. For example, the amount of copper in
given copper sulphate solution can be determined by passing current through the solution and
weighing the copper deposited at the negative electrode. Here, electric current acts as a precipitating
agent.

Gravimetric determination of a volatile constituent can be done by heating the sample and recording
the loss of weight. Gravimetric procedures are quite accurate but are lengthy and tedious

Another group of techniques was soon developed in which quantitative analysis was achieved by
measuring volume of solutions, hence it was called volumetric analysis. In this type of analysis, to
the sample solution of unknown concentration, a reagent solution of known concentration is
gradually added till the reaction between them is just complete as shown by some indicator. The
volume of the sample and reagent solutions are known, the concentration of the reagent solution is
also known so the concentration of the given sample solution can be calculated. For example, a
known volume of hydrochloric acid solution whose concentration is to be determined is taken in a
conical flask and a few drops of phenolphthalein solution are added as indicator. A solution of
sodium hydroxide of known concentration is gradually added through a burette until the solution in
the flask becomes just pink. The volume of sodium hydroxide solution added is recorded and from
this, the concentration of given hydrochloric acid is calculated. This process is called titration and
the determination is termed titrimetric determination.

Certain substances, under suitable experimental conditions, quantitatively liberate a gas. The
measurement of the volume of the liberated gas can be used as a basis for determining the substances
producing the gas. Such analytical procedures are called “gasometric methods”. For example,
semicarbazide can be decomposed with lead peroxide and the liberated nitrogen can be measured.
Knowing the volume of nitrogen, the amount of semicarbazide present in the sample solution can be
calculated.

A particular component of a gaseous mixture can be absorbed in a suitable absorbent. The decrease
in the volume of the gaseous mixture gives the volume of constituent absorbed. This method of
Dr GODAY SWAPNA

analysis is called “gas analysis”. For example, a mixture of oxygen and carbon dioxide gases can be
passed through a solution of potassium hydroxide when carbon dioxide alone is absorbed. Thus the
decrease in the volume of the gaseous mixture will be equal to the volume of carbon dioxide present
in the mixture.

Gravimetric methods along with titrimetric procedures belong to the classical methods of analysis.
Titrimetric procedures are much simpler and convenient and hence a large number of titration
methods have been worked out for determining a wide variety of inorganic and organic substances.
In direct titrimetric methods, sample solution is directly titrated with reagent solution. The indirect
procedure consists of adding a known excess of reagent and titrating back the unused reagent.

2. Classification of Methods of Quantitative Analysis

The methods of quantitative analysis can be classified from different points of view based on the
nature of material under examination, the type of method employed, the amount of desired
constituent in sample material and so on

Methods of Quantitative Analysis

Nature of material analysed – Inorganic, Organic, Biochemical

Instrumental and Non-Instrumental

Classical and physiochemical (wet anlaysis)

 The analysis can be termed organic, inorganic, biochemical etc., depending on the nature of
the material analysed.
 Another classification of quantitative analysis is non-instrumental and instrumental. The
former includes gravimetric and titrimetric procedures whereas the latter involves use of
instruments such as colorimeter, a conductivity meter or a potentiometer and so on. 
Another classification into classical and physico-chemical methods of analysis is also not
very sound theoretically although it is quite useful for the sake of convenience.
 According to this classification, gravimetric and titrimetric methods are termed chemical
methods though they are based on the measurement of physical properties, viz. weight and
volume. The methods of analysis based on such physical properties as potential,
conductance, current strength, optical rotation, etc., are generally referred to as physio-
chemical methods.
 The classical chemical analysis which consists of gravimetric and titrimetric method is also
known as wet analysis. There are certain other procedures which are based on matter-energy
interaction such as, a colorimetric determination which involves passage of light a form of
energy through solution of the substance, to be determined, which is matter.
 The methods of quantitative analysis can also be classified on the basis of the size of the
sample for a determination. The term macro-analysis is used when the determination
involves 0.1 g or more of the sample. If the amount of the sample is approximately 0.01 to
0.1 g, the method is called semi-micro and for samples weighing 0.001-0.01 g, the term
micro-method is used. Ultra-micro analysis involves samples containing less than 0.001g of

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material; some authors have used the term sub-micro analysis also. Certain procedures have
been described for analysing quantities smaller than those handled in ultra-micro analysis;
these constitute what is known as nano analysis.
 According to another classification, macro-methods are those in which sample contains more
than 2 milliequivalents (meq) of the material and 0.1 N solutions are employed in the
determination. In semi-micro methods, the sample contains about 1 meq of substance and
0.05 to 0.1 N solutions are used in its titrimetric determination. When the amount of the
substance is around 0.1 meq the method used is called a micro-method; this generally
involves the use of 0.01 N reagent solutions.
3. Evaluation of Analytical Chemistry:

A remarkable fact about the evaluation of analytical chemistry is that workers in other branches such as
inorganic, organic, biochemistry etc., have also contributed significantly to its growth. The difference in
approach is that whereas an analytical chemist have his main interest in the methods and techniques
themselves, other workers develop analytical procedures for their own specific problems. For example, a
workerin the field of chemical kinetics is not primarily interested in various analytical methods and
techniques but may develop a method for quantitatively analysing a substance whose concentration is to
be determined in order to obtain the necessary kinetic data. On the other hand main interest of an
analytical chemist is in the development and improvement of an analytical procedure itself and in testing
its reliability. He also studies the interference caused by the presence of other substances and attempt to
modify the procedure so that such an interference can be eliminated. It is necessary to distinguish
between an analytical chemist and an analyst. An analytical chemist carefully chooses a chemical
reaction and uses it for developing an analytical procedure taking into account various theoretical
considerations. He also studies the effect of different factors that can influence the result of the
determination. The job of an analyst is simply to follow the given instructions to perform a
determination.

As already been mentioned, the earlier methods of quantitative analysis were those involving
gravimetric procedures. Soon thereafter the technique of volumetric analysis emerged which due to its
inherent simplicity and rapidity received preference over gravimetric methods which were lengthy and
tedious. One of the earliest volumetric determinations was developed by Margueritte who titrated
ferrous iron with potassium permanganate. Such a procedure is now more appropriately called a
titrimetric procedure rather than a volumetric method because the latter is a more general term including
gasometric methods and gas analysis which are also based on the measurement of volume. Due to their
inherent simplicity titrimetric analyses have found and continue to find extensive applications. Both
direct and indirect titrimetric procedures have been worked out for the quantitative analysis of wide
variety of organic as well as inorganic compounds with the modern accent on micro and submicro
analysis.

Over the last 50 years or so there has been a growing tendency to make use of certain instruments to
achieve quantitative analysis. For example, instrumental techniques such as potentiometric,
conductometric, photometric, amperometric etc., have been applied to locate the end point in a titration
or to follow the course of a chemical reaction. It should be noted that in such cases the titrimetric
methods are not basically altered from their standard procedures, the instrument simply act as a
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Dr GODAY SWAPNA

substitute for an indicator. Recently such methods commonly known as the instrumental methods of
analysis have been increasingly used especially in the field of industrial and commercial quantitative
analysis due to their rapidity and sensitivity. But it would be a mistake to conclude that the so called
chemical methods would be totally eliminated from considerations. There are several reasons in favour
of this as discussed below:

The classical and physicochemical methods of quantitative analysis should be regarded as

complementary. Because once an instrument to be used in quantitative analysis has been properly
calibrated, it can be used with great advantage to achieve rapid analysis and due to high sensitivity of
physicochemical procedures, samples at microgram level can be handled.

The advantage of microchemical methods are saving of time, labour and material. Much of the work on
vitamins, hormones and other natural products could be done due to the development of microanalytical
methods because many of the compounds were present in microquantities

In about last 50 years there has been increasing sophistication in all areas of chemistry, physics and
biological sciences. This created analytical problems which required use of sophisticated
instrumentation for their solution. For example, in determining traces of impurities at part per billion
level or determining traces of pollutants in the atmosphere of industrial area.

4. Analytical Problems and their Solutios:

The solutions of all analytical problems, whether qualitative or quantitative, follow the same basic
pattern. This may be described under seven general headings:

(1) Selection: The selection of the method of analysis is an important step in the solution of an
analytical problem. A choice cannot be made until the overall problem is defined, and where possible a
decision should be taken by the client and the analyst in consultation. Inevitably, in the method
selected, a compromise has to be reached between the sensitivity, precision and accuracy desired of the
results and the costs involved. For example, X-ray, fluorescence, spectrometry may provide rapid but
rather imprecise quantitative results in a trace element problem. Atomic absorption spectrophotometry,
on the other hand, will supply more precise data, but at the expense of more time-consuming chemical
manipulations.

(2) Sampling: Correct sampling is the cornerstone of reliable analysis. The analyst must decide in
conjunction with technological colleagues how, where, and when a sample should be taken so as to be
truly representative of the parameter that is to be measured.

(3) Preliminary sample treatment: For quantitative analysis, the amount of sample taken is usually
measured by mass or volume. When a homogeneous sample already exists, it may be subdivided
without further treatment. With many solids such as ores, however, crushing and mixing are prior
requirements. The sample often needs additional preparation for analysis, such as drying, ignition and
dissolution.

(4) Separation: A large proportion of analytical measurements is subject to interference from other
constituents of the sample. Newer methods increasingly employ instrumental techniques to distinguish

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between analyte and interference signals. However, such distinction is not always possible and
sometimes a selective chemicalreaction can be used to mask the interference.

(5) Final measurement: This step is often the quickest and easiest of the seven but can only be as
reliable as the preceding stages. The fundamental necessity is a known proportionality between the
magnitude of the measurement and the amount of analyte present.

(6) Method validation: It is pointless carrying out the analysis unless the results obtained are known to
be meaningful. This can only be ensured by proper validation of the method before use and subsequent
monitoring of its performance. The analysis of validated standards is the most satisfactory approach.
Validated standards have been extensively analysed by a variety of methods, and an accepted value for
the analyte obtained. A standard should be selected with a matrix similar to that of the sample. In order
to ensure continued accurate analysis, standards must be reanalysed at regular intervals.

(7) The assessment of results: Results obtained from an analysis must be assessed by the appropriate
statistical methods and their meaning considered in the light of the original problem.

5. Importance of Analytical Chemistry to Various Branches of Science:

 Analytical chemistry finds wide applications not only in different branches of chemistry but
also in other physical and biological sciences and in many fields of engineering.
 Geologists use analytical procedures for analysing ground water, minerals, rocks, ores etc. In
agriculture, chemical analysis is used to determine the composition of soils, in the production
of fertilizers, insecticides and weed killers. Medical and biological research programmes
depend on chemical analysis which provides useful information that enhances our
understanding of vital processes and helps in developing medicines to cure various diseases.
 In order to safeguard public health there is constant checking of foods, drugs, cosmetics, water
supplies etc., and this is done in analytical laboratories. Waste disposals and the composition of
air in industrial areas are analysed to know the extent of harm they would cause to public health
so that necessary preventive steps can be taken.
 There is hardly a branch of national economy which does not make use of analytical
techniques. Chemical analysis is important in controlling the quality of raw materials,
intermediate and finished products. Hence to produce high quality products it is essential to
have analytical control at all the stages of technological processes. The sale of raw materials by
suppliers and their purchase by users is by analysis metallurgical products are most essential
materials of modern economy. The properties of alloys depend on its composition which is
established by analytical methods.
 A very large number of examples can be cited where analytical chemistry finds application. It
would be no exaggeration. If it is said that there is hardly a material related to modern living in
which analytical chemistry did not play some part. With the development of sophisticated
instruments, analytical techniques can now be used to determine traces of impurities at the part
per billion levels.

--------------------------*----------------------------
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Dr GODAY SWAPNA

CHAPTER 2
CLASSIFICATION OF ANALYTICAL METHODS AND
ADVANTAGES OF INSTRUMENTAL METHODS

Analytical chemistry can be divided into areas called qualitative analysis and quantitative analysis.
Qualitative analysis deals with the identification of substances. It is concerned with what elements or
compounds are present in a sample. Quantitative analysis is concerned with the determination of how
much of a particular substance is present in a sample. Another classification of quantitative analysis may
be based upon the size of the sample available for analysis. When a sample weighing more than 0.1 g is
available, the analysis is spoken of as macro; semi-micro analyses are performed on samples of perhaps
10 to 100 mg; micro analyses deal with samples weighing from 1 to 10 mg; and ultramicro analyses
involve samples on the order of a microgram.
Classical Chemical Analysis:
Quantitative chemical analysis started with the application of techniques of gravimetric procedures. In a
gravimetric determination, a known volume of sample solution is treated with a suitable reagent which
quantitatively precipitates the desired constituent present in the sample solution. The precipitate which is
of known concentration is filtered, washed, dried and weighed. For example, an excess of dilute
sulphuric acid is added to a given solution containing barium ions. The precipitate of barium sulphate
formed is filtered, washed, dried and weighed. From the weight of barium sulphate the quantity of
barium in the given solution is calculated.

Advantages offered by gravimetry are:

it is accurate and precise when using modern analytical balances

possible sources of error are readily checked, since filtrates can be tested for completeness of
precipitation
precipitates may be examined for presence of impurities
it is an absolute method.
it is inexpensive.

Another group of techniques was soon developed in which quantitative analysis was achieved by
measuring volume of solutions, hence it was called volumetric. To the sample solution of unknown
concentration, a solution of known concentration is gradually added till the reaction between them is just
complete as shown by some indicator. The volume of the sample and reagent solutions, the
concentration of the reagent solution are also known so the concentration of the given sample solution
can be calculated. For example, a known volume of hydrochloric acid solution whose concentration is to
be determined is taken in a conical flask and a few drops of phenolphthalein solution are added as
indicator.

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 A TEXT OF PHARMACEUTICAL ANALYSIS

A solution of sodium hydroxide of known concentration is gradually added through a burette until the
solution in the flask becomes just pink. The volume of sodium hydroxide solution added is recorded and
from this, the concentration of given hydrochloric acid is calculated. This process is called titration and
the determination is termed titrimetric determination.

Advantages of titrimetry are:

 Robustness of the method
 Analysis can also be automated
 Higher degree of precision
 Inexpensive
 Doesn’t require sophisticated instrumentation.

Over the last 50 years or so there has been a growing tendency to make use of certain instruments to
achieve quantitative analysis. For example, instrumental techniques such as potentiometric,
conductometric, photometric, amperometric etc., have been applied to locate the end point in a titration
or to follow the course of a chemical reaction. It should be noted that in such cases the titrimetric
methods are not basically altered from their standard procedures, the instrument simply act as a
substitute for an indicator.
Recently such methods commonly known as the instrumental methods of analysis have been
increasingly used especially in the field of industrial and commercial quantitative analysis due to their
rapidity and sensitivity.
Traditionally, instrumental analyses are divided into five categories.
The electroanalytical methods apply an electrical signal to the sample and/or monitor an electrical
property of the sample. The separative methods rely upon separation of the components of a sample
prior to measuring a property of the components.

1. Spectral Methods:
These types of method analysis use an instrument to calculate the amount of radiation that is absorbed,
emitted or scattered by the sample. If the amount of absorbed radiation is measured, the technique is
absorptiometry or absorption spectrophotometry.
If the absorbed energy is electromagnetic radiation in the X-ray, ultraviolet or visible region of the
spectrum, the subsequently emitted electromagnetic radiation is a form of luminescence termed either
fluorescence or phosphorescence. The spectral methods of analysis are listed in Table 1.
Table 1: List of the major spectral methods of chemical analysis

 Atomic absorption
 Atomic flouresence and ionisation

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Dr GODAY SWAPNA

 Flame and atomic emission

 Ultra violet -visible absorption by polyatomic species
 Chemilumiscence and electrochemilumiscence
 floursence and phosphorescence
 Infrared absorption
 Photoacusotic spectrometry
 Scaterring, Refractrometry
 Nuclear magnetic resonance
 Electron spin resonance
 X- ray methods
 Electron spectroscopy
 Radiochemical methods
 Thermal analysis

Some of the spectral methods are explained in brief here:

A. UV-VIS spectroscopy
B. IR spectroscopy
C. Atomic absorption spectroscopy
D. Flame emission spectroscopy
E. Atomic emission spectroscopy
A. UV-VIS spectroscopy
Ultraviolet–visible spectroscopy refers to absorption in the ultraviolet-visible region of electromagnetic
radiation. The absorption or reflectance in the visible range directly affects the perceived colour of the
chemicals involved. Ultraviolet and visible radiation absorption promotes electronic transitions σ→σ*,
n→σ*, n→π* and π→π*.
Interpretation and uses of ultraviolet spectra:
UV-spectrum of particular compound is recorded in conjunction with other spectral data such as infrared
to deduce its molecular structure.
Ultraviolet spectra, however, do not furnish prime information as such. It acts as complimentary or even
supplementary evidence to infrared.
B. IR spectroscopy:
Infrared Spectroscopy is the analysis of infrared light interacting with a molecule. Molecular vibrations
can occur by two different mechanisms. Quanta of infrared radiation can excite atoms to vibrate directly.
Most organic molecules are fairly large and their resultant vibrational spectra are complex.
Infrared absorption spectra
Two spectral regions are distinguished, region of group frequencies and fingerprint region. In group
frequency region, principal absorption bands may be assigned to vibration units consisting of only two
atoms. Such influences reveal themselves on careful study and offer valuable evidence as to the nature

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of neighboring atoms. Fingerprint region extends from 1400 cm-1 to 400 cm-1 . Absorption bands found
here are related to vibrations of molecule as a whole. Resulting bands are unique to a particular
molecule.
Advantages of IR-Spectroscopy:
 Non-Destructive
 Provides insights about functional groups present in compound
 Cheap and fast
C. Atomic absorption spectroscopy:
Modern atomic absorption spectroscopy was introduced in 1955 as a result of independent work of A.
Walsh and C. T. J. Alkemade.
Developing a quantitative atomic absorption method requires several considerations, including:
choosing a method of atomization,
selecting wavelength and slit width,
preparing sample for analysis,
minimizing spectral and chemical interferences, and
selecting a method of standardization.

Advantages of atomic absorption spectroscopy:

 High sample throughput
 Easy to use
 High precision
 Inexpensive technique

D. Flame emission spectroscopy:

Flame photometry is based on measurement of intensity of light emitted when a metal is introduced
into a flame. The wavelength of the colour tells us what the element is, and the colour's intensity
tells us how much of the element is present.
In early experiments, visible colour of the flame was used to confirm the presence of certain
elements in the sample, particularly alkali metals and alkaline-earth metals. Later the whole
ultraviolet and visible range was utilized using a spectrophotometer. This instrument permitted us to
select the wavelengths of the radiation and measure its intensity with considerable accuracy.
The spectrophotometric technique has proven to be one of the most reliable and easily used
techniques for the determination of concentrations of sodium, potassium, calcium, and magnesium.
Flame photometry is also named as flame emission spectroscopy because of the use of a flame.
Advantages of Flame emission spectroscopy:
 High sensitivity and high reliability for determination of elements in first two columns of
periodic table.
 In addition to the determination of metals, it can be applied to non-metal analysis by
utilizing the infrared region of the spectrum
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Dr GODAY SWAPNA

 Flame photometry is a simple, rapid method for the routine determination of elements that
can be easily excited.

E. Atomic emission spectroscopy:

In emission spectroscopy, a sample is excited by absorbing thermal or electric energy and the radiation
emitted by the excited sample is studied for both qualitative and quantitative analysis. Most of the
spectroscopic techniques are related to molecules but emission spectroscopy is related to atoms.
Therefore, this technique is used as a method of elementalmetal analysis. With it, all metallic elements
can be identified and quantitatively determined in very low concentrations, as can be metalloids, such as
arsenic, silicon and selenium. Solids, liquids or gases can be analyzed quite easily by this method.
Advantages of Atomic emission spectroscopy:
 Regarded as most reliable method for elemental quantitative analysis available at present.
 The method can be used for quantitative analysis of about seventy elements at concentration
level as low as 1 ppm.
 High sensitivity.

Electro analytical methods:

The electroanalytical methods are divided into categories. During analysis with most of the
electroanalytical methods, electrical contact with the sample is completed by dipping at least two
electrodes into the solution.
Amperometry is based on the control potential between the two electrodes and the current is measured.
Potentiometry is based on the controlled current between two electrodes and the potential is measured.
Various electroanalytical methods are:
 Amperometry
 Potentiometry
 Conductometry
 Coulometry
 Voltametry
 Electrogravimetry

During electroanalytical studies using coulometry and electrogravimetry, either a potential or a current
can be applied to the electrodes in the solution. The measurement of quantity Q of electricity that is
consumed during an electrochemical reaction is coulometry. When weight of a reaction product after an
exhaustive electrolysis is calculated, the method is electrogravimetry. In Conductometry, electrical
conductance of solution is calculated. Conductance is defined as the inverse of the electrical resistance.
Normally an alternating electrical potential is applied to electrodes during the measurement. In
voltammetry potential is applied to one of the electrodes while the current flowing through the electrode
is measured. During the current measurement the potential is varied in some predetermined manner.
Polarography is the series of voltammetric methods in which the electrode to which the potential is
applied has a constantly renewable surface. The most common electrode used for polarographic
measurements is dropping mercury electrode.
3. Separative Methods
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In the process of separation, the components can be individually assayed either qualitatively or
quantitatively. Among the non-instrumental separative methods are distillation, extraction, precipitation,
filtration, osmosis and reverse osmosis.
Chromatography is a technique in which a mixture is separated into its basic components. The sample is
placed on the edge of the stationary phase (a solid or liquid) and a mobile phase (a liquid or gas) is
admitting to flow over the stationary phase. Strong components sticks to the stationary phase are swept
less rapidly than weakly adhere. Result causes separation of components.
Chromatography is easily categorized into following:
1. liquid chromatography
2. gas chromatography
Depending upon the state or nature of the mobile phase. Some of the chromatographic methods are
explained in brief here:
 Paper chromatography
 Thin-layer chromatography
 Gas chromatography
 High performance liquid chromatography
A. Paper chromatography:
Paper chromatography is used to separate colored chemicals or substances. It is primarily used as a
teaching tool, having been replaced by other chromatography methods.
Apparatus:
Apparatus required for paper chromatography consists of a support for paper, a solvent trough, and an
air-tight chamber. Size of chamber may vary from an ordinary test-tube to large aquarium depending on
size of paper. Sample is applied prior to dipping into eluting solvent as a small spot.
Methods of detection commonly used are:
 inherent visible colors of components,
 reactions with color-producing reagents,
 ultraviolet absorbance,
 infrared absorbance,
 fluorescence,
 radioactivity,
 bioautography, or
 extraction and further chemical or physical tests.

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Dr GODAY SWAPNA

Test reagents are applied by spraying, immersing, or by exposing to vapors. Bioautography involves
placing paper in contact with culture medium followed by examination of growth of bacteria along
paper strip.
Advantages of paper chromatography
 Requires very less quantitative material.
 Cheaper compared to other chromatographic methods.
 Both unknown inorganic as well as organic compounds can be identified by paper
chromatography method.
 Doesn’t occupy much space compared to other analytical methods or equipments.

B. Thin – layer chromatography

Operations performed in TLC are essentially same as in paper chromatography. Instead of paper, thin
layer adsorbent supported on a glass or plastic plate is used.
Nature of layer: Commonly used adsorbents are silica gel, alumina, diatomaceous earth, and powdered
cellulose. These materials can be combined with binder to make a cohesive layer.
Preparation of layer: Layers of 0.15 to 2.0 mm thickness are satisfactory. Layers are produced by
spreading film of an aqueous slurry of adsorbent over entire surface. Slurry must be neither too thick
(viscous) nor too thin, or it will not spread properly. In commercial spreading machines a slotted trough
travels over glass and deposits a uniform layer. Binder requires about 30 minutes to "set". The edges
should be cleaned and plates stored in a cabinet. Pre-coated TLC plates (glass or plastic) are available
from many laboratory supply firms.
Methods of Development and detection: Procedure must be conducted in closed chamber. In order to
detect spots, iodine vapor is used. Another detecting reagent is a spray of sulfuric acid. There are a host
of reagents available, and, of course, spot can be scraped off, eluted, and investigated by any available
method.
Advantages: Compared to paper chromatography, thin-layer is more versatile, faster, and reproducible.
C. Gas chromatography
Basic GLC apparatus
A gas chromatograph requires a completely closed system. Components of GC are shown in Fig.3.
Carrier gas passes through pressure regulators which control flow rate through apparatus. Sample is
introduced into heated chamber either through silicone rubber septum or by sampling valve. Carrier gas
carries sample components through column where they are separated. A thermostated oven is provided
for column, injector, and detector. There are innumerable variations of basic components of gas
chromatograph.
Carrier gas:
Common carrier gases are helium, nitrogen, hydrogen, and argon. These gases are relatively
inexpensive, and are not hazardous to handle. Choice of carrier gas is usually based on availability in
high purity grade. Thermal conductivity detectors work best with hydrogen or helium. Helium is popular
choice for analysis. Flow through column is caused by difference in pressure between inlet and outlet.
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Pressure regulating valves maintain an inlet pressure, Pi outlet pressure, Po, is normally atmospheric
pressure. On some instruments, flow controllers automatically vary Pi to maintain constant flow rate.
As gases are compressible, complications arise in determining flow rate and volume of gas passing
through column.
Sample introduction:
It is important to introduce sample in shortest time and in smallest volume possible. Sample chamber
may be heated for rapid vaporization of liquid samples. In some instruments, sample is injected directly
into column at inlet. Size of sample is dictated by several factors:
 amount available,
 capacity of the column, and
 sensitivity of detector
Ordinary chromatograph can handle liquid samples in range of 0.1 to 10 µl and gaseous samples in
range from 1 to 10 ml.
Column:
There are two types of columns, packed and open tubular. Packed columns are easier to fabricate, less
expensive, last longer, have higher capacity. Open tubular columns have less pressure drop. Packed
columns are usually 1 to 20 m long. Open tubular columns are usually 10 to 50 m long. Columns are
bent in U- or W-shape or coiled to fit oven. Short columns are often made of glass. Longer columns are
made of copper, aluminum, or stainless steel.
Detectors:
Separations performed in column must be sensed and recorded. Detector must ignore large amount of
carrier gas and find trace amounts of sample components contained therein. Requirements for detector
are:
 low limit of detection,
 linear response over an extreme range of concentrations,
 uniform response to all possible substances,
 simple calibration,
 short response time,
 small internal volume,
 low noise,
 long term stability;
 it must be simple,
 inexpensive,
 robust, and
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Dr GODAY SWAPNA

 safe to operate
Advantages of gas chromatography:
 High resolution power compared to other methods.
 High sensitivity when used with thermal detectors.
 Relatively good accuracy and precision.
 Separation and analysis of sample very quickly.
 Sample with less quantity is also separated.
D. High performance liquid chromatography
This apparently simple expedient, combined with a number of other improvements in apparatus and
technique, has evolved into a new practice of liquid chromatography which is competitive with gas
chromatography in speed and resolution of complex mixtures. HPLC column consists of packing
materials.
Packing Materials: Various types of materials such as pellicular beads (covered with thin layer of
porous material), silica beads are used for packing the column. Another type of chemically bonded
packing utilizes silicone polymers which are more stable because of their three-dimensional cross-linked
structure.
High Pressures: Columns packed with small particles require high inlet pressures in order to give a
reasonable flow rate. Pressures up to 10,000 psi are not difficult to handle in the small columns used (2
to 3 mm diam.). Improved, pulse-free pumping systems are incorporated in modern liquid
chromatographs.
Detectors: The smaller columns and faster flow rates place rigid requirements on the detection system.
Flow-through detectors with low dead volumes and high sensitivity are a necessity. Two types of
detectors are currently popular. One is based on a flow-through micro-cell placed in an ultraviolet
spectrophotometer.
A second popular detector, the differential refractometer, continuously monitors the difference in
refractive index between the pure mobile phase (reference stream) and the column effluent.
Advantages of high-performance liquid chromatography:
 Widespread applicability
 Greater reproducibility
 High resolution
 Columns can be re-used
Electrophoresis is the separative method that takes advantage of the relative mobility of ions toward an
electrode of opposite charge (to the ion) and away from an electrode of similar charge.
The buffered solution through which the ions normally travel is either supported by porous paper or is in
a gel.

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Mass spectrometry can be used either alone or in combination with some other analytical technique,
such as gas or liquid chromatography.
4. Radioanalytical methods:
Radioanalytical chemistry focuses on the analysis of sample for their radionuclide content. Various
methods are employed to purify and identify the radio-element of interest through chemical methods and
sample measurement techniques.
The importance of radio-analytical chemistry spans many fields including chemistry, physics, medicine,
pharmacology, biology, ecology, hydrology, geology, forensics, atmospheric sciences, health protection,
archeology, and engineering.
Advantages of radio-analytical methods:
 Good accuracy
 Adaptability to wide variety of applications
They minimize or even eliminate the need for separations that are required in other analytical methods
6. Thermoanalytical methods
Thermal analysis is a branch of materials science where the properties of materials are studied as they
change with temperature. Several methods are commonly used:
 Differential thermal analysis (DTA)
 Differential scanning calorimetry (DSC)
 Thermogravimetric analysis (TGA)
Advantages of Thermo-analytical methods:
 High resolution
 Rapid response time
 Easy measurement on samples of different configurations
-------------------------*---------------------------

CHAPTER -3
SOLUTION REACTIONS

Introduction
Many of the chemical reactions happen in a solution; water is the most commonly used solvent.
Different other liquids can also be employed as solvent instead of water. Therefore, to know about the
factors influencing the chemical reactions is very necessary.

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Dr GODAY SWAPNA

Chemical Equilibrium Hydrogen iodide is formed when hydrogen and iodine vapours combine after
heating together in a closed container at 450⁰C. However, some hydrogen and iodine in this experiment
is left uncombined and their rate of combination is independent of duration of experiment. Again if
heating of pure hydrogen iodide is done in similar conditions then hydrogen and iodine are again formed
by the decomposition of hydrogen iodide, but here also rate of decomposition is independent of duration
of heating and some amount of hydrogen iodide is left unchanged. This chemical reaction is reversible
in nature.
H2 (g) + I2 (g)--------- 2HI (g)
The following chemical reaction in liquid phase is also reversible in nature. In this reaction esterification
takes place between ethanol and acetic acid. Products which are formed in the reaction are ethyl acetate
and water. Due to the conversion of ethyl acetate when heated with water to acetic acid and ethanol
again, esterification process remains incomplete.
C2H5OH +CH3COOH------- CH3COOC2H5 + H2O
After sometime all reversible reactions reach a state of chemical equilibrium. Composition of
equilibrium mixture in this state remains constant, if temperature (also pressure in case of some gaseous
reactions) remains constant. If constant conditions are maintained then same state of equilibrium can be
attained from any of the two directions of a given reaction which is reversible in nature. In equilibrium
state, two reactions which are opposite to each other occur at an identical rate and therefore the system
remains in dynamic equilibrium state.

Composition of a particular system in equilibrium can be changed by altering conditions of a given

reaction. Therefore, it is essential to take into consideration the influence of changes in (i) temperature
(ii) pressure (iii) concentration of various reagents. The principle of Le Chatelier-Braun is stated as: 'If a
constraint is applied to a system in equilibrium, the system will adjust itself so as to nullify the effect of
the constraint'. Explanation of the above mentioned factors can be easily done on the basis of the
principle of Le Chatelier-Braun.

(i) Temperature: Formation of ammonia is reversible in nature:

N2 (g) + 3H2 (g) 2NH3 (g)

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In the above process, heat is evolved in the forward reaction, and is termed as exothermic process;
however, the reaction in reverse takes place by absorption of heat and the process is endothermic. If
temperature of the system at an equilibrium state is raised, then the process where absorption of heat
takes place is favoured resulting in decomposition of ammonia.

(ii) Pressure: In reference to equilibrium system of hydrogen iodide, stoichiometric coefficients

of the molecules on both sides of an equation for a given reaction are equivalent, and the
volume remains unchanged when reaction takes place. Thus, if system’s pressure is doubled,
which halves the total volume, then both equation sides get affected equally, and therefore
equilibrium mixture’s composition doesn’t change.
(iii) Concentration of various components: If addition of hydrogen is done to equilibrium
mixture, then hydrogen iodide decomposes thermally, it is observed that there is presence of
more hydrogen iodide when equilibrium is restored.

Law of mass action

Law of Mass Action states that: 'The velocity of a chemical reaction is proportional to the product of
the active masses of the reacting substances'. 'Active masses' of the reactants were given in moles per
litre. A mathematical equation for equilibrium condition in a reversible reaction can be obtained by
applying this law to homogeneous systems. For a reversible process which takes place at constant
temperature:

A+B ------- C+D

Conversion rate of A and B is proportional to concentration of A and B respectively,

r 1 = k 1 x [A] x [B]

where k1 is known as rate coefficient or rate constant, and square bracket is used to denote
concentration in mol L -1 of the compound. Rate of conversion of C and D can be presented in the
same way:

r 2 = k 2 x [C] x [D]

At equilibrium state, conversion rate of A and B and rate of conversion of C and D are equal:

r1 = r2

or

k 1 x [A] x [B] = k2 x [C] x [D]

[C] x [D]/[A] x [B] = k1/k2 =K

K is equilibrium constant of a given reaction at a particular temperature. General expression for a

reversible process can be given by:

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Dr GODAY SWAPNA

p 1A 1 + p 2A 2+ p3 A3+ ---------q 1 B 1 +q2 B2 + q3 B3+ ...

where p1, p2, p3 and q1, q2, q3 are stoichiometric coefficients of reactants, equilibrium condition
can be expressed by:

[B 1] q1 x [B 2] q2 x [B 3] q3/ [A 1] p1 X [A 2] p2 x [A 3] p3 ....... = K

When a reversible reaction attains state of equilibrium, then at a fixed temperature, product of
concentration of the product/products divided by product of concentration of reactants and each
concentration is raised to a power equal to stoichiometric coefficient of that specific compound in
the equation for the reaction is constant.

Mainly three factors affect the reactions in a solution. They are: (i) nature of solvent; (ii)
temperature; and (iii) catalysts.

Nature of solvent: In aqueous solutions, reactions usually proceed rapidly as there is interaction
between ions. Reactions within molecules in a solution, such as formation of ethyl acetate when acetic
acid and ethanol reacts, are usually time taking. Hence, for convenience a solvent can be classified as an
ionising solvent if it possesses a tendency to yield a solution where the solute is ionised. A non-ionising
solvent is one (methanol, ethanol, esters, ethers and hydrocarbons) which produces a solution in which
the solute doesn’t ionise. Examples of ionising solvents are ammonia, hydrogen chloride, water, acetic
acid, sulphur dioxide, amines and bromine trifluoride. Of these solvents, first four are capable of
producing hydrogen ions, as for example:

H2 O ---- H3O+ + OH-

These four solvents are termed as protogenic solvents, while sulphur dioxide is a nonprotonic
solvent as it doesn’t contain hydrogen.

Temperature: With the increase in temperature, rate of the reaction also increases. In some cases, the
solution is heated so that the reaction occurs rapidly. For example, when acidified oxalate solution is
titrated with KMnO4, the reaction is very slow. But heating of solution to about 70°C prior to the
addition of permanganate solution, makes the reaction occur instantaneously.

Presence of catalysts: Catalysts can increase the rates of some reactions to a great extent. Catalyst can
alter reaction rate without any net change in itself. Conversion of large amounts of reacting species can
be easily done with very small quantity of catalyst.

Electrolytic dissociation: Electricity is conducted by aqueous solutions of some salts of strong acids as
well as bases. These are known as electrolytes. However, some solutes, for example ethylene glycol
produces solutions showing conducting ability which is slightly different from water: such solutes are
known as non-electrolytes.

Salts: If a salt is dissolved in a solvent whose dielectric constant is high, or when they are heated to
melting point, the crystal forces get weak and salt dissociates into ions which existed previously, due to
this the resulting solutions possess fair amount of conductivity.

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These substances are known as strong electrolytes. Few salts give solutions showing significant amount
of conductance, but less than solutions of strong electrolytes having a similar concentration. These types
of solutes are known as weak electrolytes.

Acids and bases: An acid dissociates to form hydrogen ions as the only positive ions when dissolved in
water:

HNO3 ------ H + + NO3

In aqueous solution, hydrogen ion does not exist in free-state and therefore it combines with water
molecules to form hydroxonium ion.

HNO3 + H2O -------- H3O + + NO3 –

Ionization is due to the tendency of ions of hydrogen in a free state to combine with water molecules
forming hydroxonium ions.

Polyprotic acids: Acids which ionise in stages are known as polyprotic acids. For example, in H2SO4,
one atom of hydrogen is ionised completely:

H2SO4 + H 2O ------ H3O + + HSO4

Second atom of hydrogen is only partially ionised, except in dilute solutions:

H2SO4 + H2O -------- H3O + + SO4 2-

Ionisation stages are primary, secondary and tertiary ionisations. Extent of dissociation of acids of the
type of acetic acid is small. The former acid is known as strong acid (hydrochloric, nitric and sulphuric
acids), and latter is termed as weak acid (acetic acid, boric acid and hydrogen sulphide).

Originally, definition of a base was given as that, it is a substance which dissociates to form hydroxide
ions as the only negative ions when dissolved in water. Thus sodium hydroxide gets completely
dissociated in aqueous solution and is a strong base.

NaOH -------->Na+ + OH

Aqueous ammonia solution produces a very little amount of hydroxide ions in aqueous solution and is a
weak base.

NH3 + H 2O --------->NH4 + + OH

Bronsted-Lowry theory: This theory was proposed by J. N. Bronsted and T. M. Lowry. The theory
stated that, acids are substances having a tendency to lose proton while bases are substances having a
tendency to add on a proton.

Acid -------- Proton + Conjugate base

A ---------- >H + + B (i)

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Dr GODAY SWAPNA

As free protons are unable to exist in a solution in a measurable concentration, reaction doesn’t occur
unless addition of alkali is done which can take the proton furnished by an acid. If both the equations are
combined,

A1------- B1 + H+ and B2 + H+ --------- A2

We get,

A1 + B2 ------- A2 + B1 (ii)

A1 – B1 and A2 – B2 are conjugate acid-base pairs. This expression represents transfer of a proton from
A1 to B2 or from A2 to B1.

In aqueous solutions, a Bronsted-Lowry acid, A is said to be strong if equilibrium in the above reaction
is practically complete to the right so that [A] is almost zero.

A + H2O ---------- > H3O + + B

For a strong base, equilibrium concentration of base [B] other than hydroxide ion is near to zero. Thus
arrangement of acids can be done in series according to their relative tendencies to combine with base:

HCl + H2O -------- H3O + + Cl

Acid Base Acid Base

The above reaction is essentially complete for all strong acids such as HCl. Reactions of weak acids,
proceed only slightly to right:

CH3COOH + H2O ------------- H3O + + CH3COO-

Acid Base Acid Base

Hydrated proton is a typical strong acid of water system and a conjugate base doesn’t have a major role
if it is weak. Strength of a conjugate base may vary inversely as strength of the respective acid. Basic
ionisation constant of a conjugate base (KB.conj.) is equal to Kw/ KA.conj. where Kw is ionic product
of water.

G. N. Lewis related properties of an acid to accept pairs of electron and base as a donor of pairs of
electrons, to form covalent bonds without regarding the involvement of protons. Theory of complete
dissociation of a strong electrolyte in an aqueous solution was further explained by Debye, Huckel and
Onsager.

---------------------------*-------------------------------

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CHAPTER 4
LABORATORY OPERATIONS AND GOOD LABORATORY
PRACTICES
Measurements are done in an analysis to obtain facts. These measurements include weighing on an
analytical balance, analysis bycalibrated glassware and apparatus such as ovens, furnaces and filters.
Basic requirements for an analysis are given in the following section.

Laboratory notebook for recording data:

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Dr GODAY SWAPNA

Cleanliness is very important in a laboratory. A lab notebook should also be kept neat with proper
results. The data of an experiment should be recorded at that time only when the experiment is being
performed and it should be dated. By doing so, a lot of time in future can be saved and the analyst is
more organized in performing the experimental work with a great ease.

If the analyst immediately records the data, possible errors in measurements can easily be detected. The
chances of losing important data are also reduced if the recording of results is done in a notebook rather
on pieces of paper.

Electronic notebooks:

Modern equipment software has made possible to gather and analyse data directly from the signal
obtained from the instrument. Proper calibration of software and apparatus is mandatory prior to any
analysis.

Laboratory Materials:

The most popular and widely used material for analysis in a laboratory is borosilicate glass. It is
employed in making beakers, pipets, flasks and burettes. Borosilicate glass can withstand high
temperatures easily and so is a popular choice for the analyst.

Titration:

Titration is done with solution of the sample contained in a flask. While adjusting the stopcock with the
left hand the flask is shaken with the right hand. Magnetic stirrer is a more effective way for mixing the
contents in the flask. Initially when indicator is added to the solution in the flask, changes in colourare
observed owing to local excesses; but it gradually returns back to the original colour as the titrant
getsmixed through sample solution. When the end point is near, reversion to the original colour becomes
slower, since the solution at this point of time must be stirred more effectivelyfor consumption of the
titrant. The process of titration at this point should be stopped.

Tolerances and Precision of Glassware:

Tolerances, or absolute errors, for various volumetric glassware has been specified by National Institute
of Standards and Technology (NIST). Tolerance for volumes greater than about 25 mL, should be within
l part per thousand (ppt) relative. Letter "A" on a volumetric glasswareis an indication that the particular
glassware complies with class A tolerances. Volumetric glasswareswhich meet NIST specifications are
much costly than the simple glasswares. Cheap glassware although they are financially suitablemay have
higher tolerances than those prescribed by NIST.

Calibration of Glassware:

 Calibration of volumetric flask:

For calibration of a volumetric flask, it should be first cleaned thoroughly of any dirt then weighed
properly and dried in an oven and stoppered. After that the clean and dried flask is filled with

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 A TEXT OF PHARMACEUTICAL ANALYSIS

distilled water up to the given mark. While filling the flask care should be taken to avoid adherence
of water droplets on the neck of the flask. If droplets are present, then they should be wiped out with
clean tissue paper. The volumetric flask and distilled water filled in the flask both should be at room
temperature. The filled flask is then weighed, and then temperature of water to 0.1o C is recorded.
Increase in the weight of the flask is the weight in air of water filledin the flask.

 Calibration of pippet:

For calibration of a pipet, an Erlenmeyer flask is cleaned and then weighedhaving a rubber stopper
or weight of a clean weighing bottle stoppered with cap is taken. The selection of the glassware
depends on volume of water to be weighed. The pipet is filled with distilled water and it isdelivered
into the flask or bottle, by employing proper technique of pipetting. The flask or bottle
isimmediately stoppered to prevent loss by evaporation. The container is then again weighed to get
the weight in air of distilled water transferred by pipet.

 Calibration of burette:

Calibration of a burette is same as that for a pipet, except that for a burette several volumes need to
be delivered.

Glassware selection for analysis

As in weighing procedures, there will be circumstances where it is necessary to know, with great
accuracy, thevolumes of reagents which are measured or delivered, and many others where only
relative measurements are needed. For preparation of a standard solution of 0.1 M HCl, it can't be
accomplished by taking an accurate volume of acid and its dilution to a known volume as the
commercial acidconcentration is not known properly. Therefore, an approximate solution can be
prepared which is standardized, molarity of commercial acid is approximately 12.4 M, and for
preparation of 1 L of asolution of 0.1 M, 8.1 ml of HCl is taken and diluted. For this purpose, a 10-
mL graduated cylinder or 10 ml pipet is sufficient, and dilution of the acid can be done in an
ungraduated l-L bottle. For dilution of a stock standard solution with accuracy, a transfer pipet must
be employed and dilution should be done in a volumetric flask.

Standard Base Solutions:

Sodium hydroxide is generallyemployed as the titrant when there is a need of base.

NaOHcontainsappreciablequantities of H2O and Na2CO3, and therefore it cannot be employed as a
primary standard. To achieve accuracy in an analysis, Na2CO3needs to be removed from NaOHas a
buffer is formed by their reaction. A decrease in sharpness of the end pointis observed due to the

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Dr GODAY SWAPNA

buffer. Additionally, erroneous result is obtained if sodium hydroxide is standardized by the help of
phenolphthalein end point, and after that methyl orange end point is employed in titratingsample.
Na2CO3is not soluble in nearly saturated NaOH and is removed by dissolution of the weighed
NaOH in a volume of water equivalent to its weight in grams. Insoluble Na2CO3is allowed to settle
down for many days, and then supernatant is decanted carefully, or it is filtered by the help of a
Gooch crucible equipped with an asbestos mat. Filtration with gooch crucible generally not preferred
as asbestos is carcinogenic in nature.

CO2 from air gets dissolved in water. In routine analyses which do not requiremuch accuracy,
carbonate or CO2 impurities in water result in an error which is negligible. For highest degree of
accuracy, removal of CO2 should be done from all water employed for preparation of solutions for
acid-base titrations, especially the alkaline solutions. This can be done easily by boiling and then
after cooling under cold-water tap. Sodium hydroxide is generally standardized by titration with a
weighed amount of primary standard potassium acid phthalate (KHP). Storage of NaOHsolution
should be done in a plastic bottle to avoid absorption of CO2 from the atmosphere.

standard Acid Solutions:

Hydrochloric acid is usually used as a titrant for titrating bases. As chlorides are soluble someside
reactions are possible by the use of this acid. This acid can be handled conveniently. This acid is not
a primary standard, and therefore an approximate solution is prepared by dilution of the concentrated
acid. Primary standard Na2CO3 isgenerallyemployed for standardization ofHCl solutions. Its
demerit is that the end point obtained is not so sharp unless methyl red, methyl purple, and so forth is
employed as indicator and the solution is boiled at the end point. A modified methyl orange end
point can be employed without boiling, but this is not sharp. Another limitation is that it has low
formula weight. If there is availability of standardized solution ofNaOH, thenHClis standardized by
titration of an aliquot with the standardized NaOH. HCl is titrated with base, rather than the other
way around, to keep absorption of CO2minimum in the titration flask. Phenolphthalein or
bromothymol blue can also be employed as indicator.

Apparatus for handling and treating samples:

There arenumerousequipments available for use in analytical procedures.

Blood Samplers

For collection of blood samples syringes are generally used. Stainless steel or aluminium needles
are usuallyemployed with glass or plastic syringes. These generally present no issue of
contamination. However, specific precautions may benecessary for particular trace element
determination.

Oven:

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An oven is employed to dry samples before taking their weight. These ovens are properly ventilated.
The temperature inside the oven is about 110°C, but temperatures of 200 to 300°C can be achieved.

Hoods:

When evaporation of solutions is to be done a fume hood is generally used. When perchloric acid or
its solution is evaporated fumes are generated, these fumes either are collected, or evaporation
process isdone in fume hoods. Vertical laminar flow stations are generally preferred.

Wash Bottles:

A wash bottle is very helpfulin a laboratory. It can be used to quantitatively transfer precipitates and
solutions and in washing precipitates. Wash bottles are available in different shapes and sizes.

Centrifuges and Filters:

A centrifuge is of great help in the clinical laboratory, where blood needs to be separated into
different fractions such as serum or plasma, and proteins are to be separated by precipitation
followed by centrifugation. Filters for filtration of precipitates are of different types. Gooch crucible
and porcelain filter crucible are some examples.

Filtration:

Rate of filtration can be increased by properly fitting the filter paper. Filter paper is folded in form of
a cone. After placing folded paper infunnel, it is wetted with distilled water. The process of filtration
is started immediately.

Ignition of Precipitates-Gravimetric dtermination:

Before ignition of a precipitate in a porcelain filter crucible, moisture is removed first at a very low
temperature. A muffle furnace orburner can be used for ignition purposes. If a burner is employed,
filter crucible iskept in a porcelain or platinum crucible to avoiddiffusion of gases present in
flamethrough pores of filter. The crucible should be dried to constant weight before drying the
precipitate in it. Crucible needs to be ignited to constant weight if the obtained precipitate is to be
ignited. The crucible is cooled in desiccator for some time before weighing. Red-hot crucibles are
cooled below redness beforekeeping them in desiccator.

Sampling

Sampling plays a very important role in an analysis. Accuracy of measurements is affected greatly
by sampling. If sampling is not done in a proper manner, the whole of the analysis is affected
adversely. Many professional agencies have providedspecific instructions for samplinge.g.,
American Society for Testing and Materials (ASTM), Association of Official Analytical Chemists
International (AOAC International) and American Public Health Association (APHA). With the help
of experience and statistics, sampling can be done accurately. In sampling, a representative sample
known as the gross sample is obtained from the vast amount of material. This gross sample is
reduced to a small size which is easy to handle. This is known as the sample. From this sample, an

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Dr GODAY SWAPNA

aliquot, or portion is taken for analysis. This portion is known as the analysis sample. Analysis is
repeated several times on the same sample by taking different aliquots.

Drying operations and preparation ofanalyte solution:

Sample drying: Solid samples generally contain some amount of adsorbed water. With inorganic
materials, sample isusually dried before taking weight. This is done by keeping it in an oven at about
105 to 110°C for 1 to 2 hours. Water which is entrapped within the crystal, can only be removed at a
higher temperature. Drying of substances which are heat sensitive can be done in a desiccator; use of
a vacuum desiccator saves much time.

Dissolution of samples:

Prior to measurement of an analyte, sample alteration is usuallyrequired to get the desired analyte
into solution or, for biological samples, to free them from interfering materials, such as proteins.
Sample preparation can be done in following two ways: one procedure is by total destruction of
sample matrix and the other procedure is non-destructive or only partially destructive. The former
procedureis usually employedif the desired analyte is inorganic or it can be easily converted to an
inorganic derivative. The latter procedure is employed if analyte to be analysed is an organic
substance.

Dissolution ofinorganic solids:

Strong mineral acids can be used for dissolving many inorganics. HClbeing a good solvent dissolves
easily many metals. Nitric acid can dissolve many metals, alloys and "acid-insoluble" sulfides.
Perchloric acid is a very effective oxidizing acid. It can dissolve many metals and can also
destroyremains of organic material. However, this acid must be employed with extreme precautions
as it may react explosively with different oxidizing substances.

Inorganic Analysis by destruction of organic matter:

1. Dry Ashing:

Simple dry ashing is the most common technique. Generally, a porcelain crucible is employed.
Platinum crucibles are usually employed for lead. If an oxidizing substance is added to sample,
ashing efficiency can beincreased. Magnesium nitrate is an example which is helpful in enhancing
ashing efficiency.

2. Wet Digestion:

Wet digestion employing mixture of nitric and sulfuric acidsisoften used. Generally, a small
quantity (e.g.,5 mL) of H2SO4 is employed with much larger volumes of HNO3(20 to 30 mL).
AKjeldahl flask is generally used for wet digestion. Bulk of the organic material is destroyed by
HNO3, but it is not capable to destroy the last traces. Digestion is continued until a clear solution is
obtained.

Preparing samples with help of microwave:

37
 A TEXT OF PHARMACEUTICAL ANALYSIS

Microwave ovens are generallyemployed for fast and complete drying and aciddecomposition of
various samples. Digestion with the help of microwave has many advantages such as the time
required for dissolution is very less and low blank levels due to small quantities of reagents needed.

Protein FreeFiltrates (PFF):

Proteins present in biological samples pose interference in many determinations and therefore they must
be removednondestructively. Different reagents are helpful in precipitation of (coagulate) proteins.
Trichloroacetic acid (TCA)and bariumhydroxide plus zinc sulfate are some of the reagents commonly
used for this purpose. Measured volume of the sample is generally treated with reagent. After
precipitation of protein, filtration of the sample is done through filter paper without washing or else its
centrifugation is done. An aliquot is then taken and analyzed.

Good laboratory Practices:

Good laboratory practices (GLP) play a very important role in an analysis and assure that the results of
an analysis are correct within limits specified by different agencies.

Basic elements for GLP:

The most crucial point is thatmanagement of laboratory and analysts should employ common sense in
judging what quality assurance proceduresneed to be implemented, based on the goal of analysis,
experience, methods available, time and cost constraints, and the like. If an analyst follows GLP
specifications, then he/she will have more confidence in the results obtained. GLPs can be defined as
operating proceduresprescribed by a given agency that are considered to be necessary to ensure quality
in the results obtained. They all contain two elements: standard operating procedures (SOPs) and a
quality assurance unit (QAU). Standard operating procedures describe the actions performed in a
laboratory. Examples are sample handling and preparation, analytical method, instrument / maintenance,
archiving (record keeping), and the like. The quality assurance unit is usually independent from
laboratory-and is answerable to manager of institution with which the-laboratory is affiliated.QAU is
mainly responsible for implementation of quality procedures and evaluating them on a regular basis; this
includes laboratory audits from time to time.

Accreditation of laboratory

Another form of assessment is laboratory accreditation by formal institution or government agency.

This is usually voluntary but may be needed for labs dealing with regulatory measurements. In
accreditation, an authoritative agencyprovides formal recognition that the lab is capablefor carrying out
specific analysis. Accreditation verifies that good laboratory practice policies are adhered to. A number
of government organizations have specified guidelines for validation of developed method. Some of
them are: International Organization for Standardization, International Conference on Harmonization,
Organization for Economic Cooperation and Development, Food and Drug Administration
Environmental Protection Agency andOffice of Enforcement and Compliance: Laboratory Data.

38
Dr GODAY SWAPNA

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39
 CHAPTER 5
STEPS INVOLVED IN QUANTITATIVE ANALYSIS

 Methods of analysis aredivided into two broad areas called qualitative analysis
andquantitativeanalysis. Qualitative analysis deals with the identification of substances. It is
concerned with what elements or compounds are present in a sample. Quantitativeanalysis
measuresamount of a particular substance present in a sample. The substance determined is
called as the desired constituent or analyte. If the analyte constitutes more than about 1% of the
sample, it is considered a majorconstituent. It is considered minor if it amounts to 0.01 to 1% of
the sample. Finally, a substance present to the extent of less than 0.01% is considered a trace
constituent.
 Another classification of quantitative analysis may be based upon the size of the sample
available for analysis. the subdivisions are not clear-cut, but merge imperceptibly into one
another and are roughly as follows: when a sample weighing more than 0.1 g is available, the
analysis is spoken of as macro; semimicro analyses are performed on samples of perhaps 10 to
100 mg; micro analyses deal with samples weighing from 1 to 10 mg; and ultramicro analyses
involve samples on the order of a microgram (1µg = 10-6g)

Analytical methodology
In the introductory course in quantitative analysis students deal mainly with major constituents of
macro samples. They seldom perform a complete quantitative analysis of sample.
A complete analysis actually consists of five main steps
(1) Sampling, that is selecting a representative sample of the material to be analyzed;
(2) dissolution of the sample;
(3) conversion of the analyte into a form suitable for measurement;
(4) measurement; and
(5) calculation and interpretation of the measurement. Often beginners carry out only steps 4 and 5,
since these are usually the easiest ones.
In addition to the steps mentioned above, there are other operations that may be required. If the sample
is a solid, it may be necessary to dry it before performing the analysis. An accurate measurement of the
weight of the sample (the volume if t is a gas) must be made, since quantitative results are usually
reported in relative terms, for example, the number of grams of analyte per 100 g of sample (percent by
weight). At this time we shall make some general comments on the steps in an analysis:

 Sampling:
Beginning students do not find the problem of sampling, since the samples they are given are usually
homogeneous, or nearly so. Nevertheless, they should be aware of the importance of sampling and
should know where to find proper directions when they are confronted with an unfamiliar problem. We
shall discuss briefly the sampling of solids, liquids and gases to give a general idea of the nature of the
problems involved. The process involves statistical reasoning, in that conclusions will be drawn about
the composition of the bulk sample from the analysis of a very small portion of material.
 Solids:
Sampling of coal is very difficult, and we shall use it to illustrate methods used for solid materials. The
first step in the sampling procedure is selection i.e. a large portion of coal is selected, called the gross
sample, which though not uniform itself, represents the average constituent of the entire mass. The
quantity of the sample needed depends on such factors as particle size and homogeneity of the particles.
In the case of coal the gross sample must be about 1000 lb if the particles are no greater than about 1
inch in any dimension.
There are various techniques used to obtain the gross sample. If the coal is in motion on a conveyor of
some type, a specified fraction may be continuously diverted to give the gross sample. If, on the other
hand, the coal is being pass from a car, every fiftieth cycle might be placed aside to form the sample.
After the sample selection, it is ground or crushed and uniformly mixed and reduced in size. One
method used for reducing the sample of coal involves piling it in a cone with a shovel, flattening the
cone, and dividing it into four equal parts, two of which are discarded. A mechanical device for
subdividing the sample is called a riffle. The riffle contains a row of small sloping chutes arranged so
that alternate chutes discharge the sample in opposite directions.
In this manner the sample is split into two parts automatically. In the laboratory further grinding of the
sample may be done with a mortar and pestle, so that sample can pass through a sieve of a certain mesh.
One hopes that the final laboratory sample, 1 g or so, is representative of the gross sample. The
analytical data obtained cannot be better than the care taken in the sampling procedure.
 Liquids:
If the liquid is homogeneous, the sampling procedure is simple. The process is much more difficult if the
liquid is heterogeneous. In the case of a liquid circulating in, for example, a pipe system, samples are
often taken from different points in the system. In a lake or river, samples may be taken at different
locations and at different depths. Sometimes the analyst may not wish to have an average sample for the
entire liquid system. For example, in testing the natural purification of a river contaminated with
sewage, samples may be taken at a number of places downstream from the sewer outlet.
For the analysis, sample can be collected from the large water bodies with the help of a device called
grabsampler. The main components of this device are, sample bottle inside a metal container sufficiently
heavy to force the empty bottle to the desire depth. The sample bottle is closed by a stop cork, which has
a line attached to it, and held by the person taking the sample. The device is lowered to the desired
depth, the stop cork is pulled out, and the sample bottle is filled. The sampler may have a floating ball,
which automatically seals the bottle when it is filled.

 Gases:
There is much interest todayinsampling the atmosphere because of efforts directed toward improvement
of the quality of the air we breathe. Air, is of course, a complex mixture which contains particulate
Dr GODAY SWAPNA

matter as well as numerous gaseous compounds. Its composition depends on a number of factors. Such
as location, temperature, wind and rain.
In the collection of an atmospheric sample for analysis the volume taken and the rate and duration of
sampling are important factors. The air is passed through a series of fine filters to isolate particulate
matter, and through a column of a solution where a chemical reaction occurs to trap the desired the
component. After collection on a filter, particulate matter, may be determined by chemical analysis or by
weighing. The general requirements for sampling solids, liquids, and gases may be found in general
reference works.
 Sample dissolution:
Many of the samples analyzed in the beginning course in quantitative analysis are soluble in water.
Naturally occurring materials, such as ores, and metallic products, such as alloys, must be given
special treatments to prepare their solution. While each material may present a specific problem, the
two most common methods employed in dissolving samples are (1) treatment with hydrochloric,
nitric, sulphuric or perchloric acid, and (2) fusion with an acidic or basic flux followed by treatment
with water or an acid. The solvent action of acids depends upon several factors:
1). The reduction of H+by metals more active than hydrogen: for example,
Zn(s) + 2H+→ Zn2+ + H2(g)
2). The combination of H+with anion of a weak acid: for example,
CaCO3 + 2H+ → Ca2+ + H2O + CO2(g)
3). The oxidising properties of the anion of the acid: for example,
3Cu(s) + 2NO3 - + 8H+ → 3Cu2+ + 2NO(g) +4H2O
4). The tendency of the anion to from soluble complexes with the cation of the substance dissolved: for
example,
Fe3+ + Cl- → FeCl2+
Hydrochloric acid and nitric acid are most commonly used to dissolve samples. The chloride ion is not
an oxidising agent as is nitrate ion, but it has a strong tendency to form a soluble complex with many
elements. A very powerful solvent, aquaregia, is obtained by mixing the hydrochloric acid and nitric
acid in the ratio of 3:1.
Many substances that are resistant to attack by water or acids are more soluble after fusion with an
appropriate flux. Basic fluxes such as sodium carbonate are used to attack acidic materials such as
silicates. Acidic fluxes such as potassium hydrogen sulphate are used with basic materials such as iron
ores. Oxidising or reducing substances can also be used in certain cases. Sodium peroxide, for example
is often employed as a flux.
 Preparation of the analyte solution
Before a physical or chemical measurement can be made to determine the analyte in a solution of the
sample, it is usually necessary to solve the problem of “interferences”. Suppose for example, by adding
potassium iodide and titrating the liberated iodine with sodium thiosulfate. If the solution also contains
iron (III) ion, this ion will interfere, since it also oxidises iodide to iodine. The interference can be
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prevented by adding sodium fluoride to the solution, converting iron (III) into the stable complex FeF6
3. This is an illustration of a general method in which interferences are effectively “immobilized” by
alteration of their chemical nature. A second method involves physical separation of the analyte from the
interferences. Suppose that one wishes to determine magnesium in a sample which also contains iron
(III) ion and the magnesium is to be precipitated as the oxalate. The iron will interfere, since it also
forms a precipitate with oxalate. The iron can be precipitatedas the hydroxide using ammonia at a pH of
about 6.5. The magnesium is not precipitated at this pH, and hence the interference is removed. In
gravimetric analysis the analyte is physically separated from all other components of the sample as well
as from the solvent. For example, the chloride in a sample may be determined by precipitation of silver
chloride, which is then filtered, dried, and weighed. Precipitation is one of the more widely used
techniques for separating the analyte from interferences. Other important methods include electrolysis,
solvent extraction, chromatography, and volatilization.
 Measurement
The initial step of measurement begins with by chemical, physical, or biological means. The analytical
techniques employed has led to the classification of quantitative methods into the three subdivisions,
titrimetric (volumetric), gravimetric, and instrumental. A titrimetricanalysisinvolves measurement of the
volume of a titant which is required to react with the analyte. In a gravimetric method the measurement
is one of weight; the term instrumental analysis is used rather loosely; it originally referred to the use of
a special instrument in the measurement step. Actually, instruments may be used in any or all steps of
the analysis, and frankly speaking, burets and analytical balances are instruments. Spectroscopy, both
absorption and emission, is perhaps the most widely used instrumental method. Other instrumental
methods include potentiometry, polarography, coulometry, conductimetry, polarimetry, refractometry
and mass spectrometry.
Calculation and Interpretation of the results:
After the measurements have been completed, quantitative results are mathematically manipulated, and
both qualitative and quantitative results are presented in a meaningful manner. The final step in a
quantitative analysis is calculation of the percentage of analyte in the sample. The principles involved in
such calculations are normally simple. For example, titrimetric and gravimetric methods are based on
the simple stoichiometric relationships of chemical reactions. In spectrophotometric methods the
property measured, absorbance, is directly proportional to the concentration of the analyte in the
solution. On the other hand, interpretation of the results obtained by analytical methods is not always
simple. Since errors can be made in any measurement, the analytical chemist must consider this
possibility in interpreting the results. The methods of statistics are commonly used and are especially
useful in expressing the significance of analytical data.
In most cases, two values are reported for quantitative analyses. The first value is an estimate of the
correct value for the analysis, and the second value indicates the amount of random error in the analysis.
The most common way of reporting the best value is to give the mean (average) of the results of the
laboratory assays. In specific cases, however, it is better to report either the median (central value when
the results are arranged in order of size) or the mode (the value obtained most often).
Accuracy is the degree of agreement between the experimental result and the true value. Precision is the
degree of agreement among a series of measurements of the same quantity; it is a measure of the
reproducibility of results rather than their correctness. Errors may be either systematic or random.
44
Dr GODAY SWAPNA

Systematic errors cause the results to vary from the correct value in a predictable manner and can often
be identified and corrected. An example of a systematic error is improper calibration of an instrument.
Random errors are the small fluctuations introduced in nearly all analyses. These errors can be
minimized but not eliminated. They can be treated, however, using statistical methods. Statistics is used
to estimate the random error that occurs during each step of an analysis, and, upon completion of the
analysis, the estimates for the individual steps can be combined to obtain an estimate of the total
experimental error.
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45
 CHAPTER 6
CALIBRATION OF GLASSWARE AND INSTRUMENTS
Introduction Most measurements become useful only through comparisons: a measured value by itself
can not usually be interpreted without a comparison point. Moreover, the raw data in most analytical
measurements are recorded in units of potential difference (volts, V) or current (amperes, A) that can be
achieved by many different phenomena not necessarily the quantity we need to measure. This recorded
quantity is technically called a measurand. In analytical chemistry the term analyte refers to a certain
chemical species, such as an element, ion, molecule, or radical.
Before the electrical signalsaremeasured and interpretable it is compared with a similar one measured
under same experimental conditions, but on a sample with well-defined values of the measurand. Such a
sample with known properties regarding the quality under investigation is called a standard. The higher
the signal in the analysis of the standard, the higher is the expected signal in the measurement of the
same property on a sample with a (previously) unknown value of the measurand.
The signal can increase or decrease with increasing value of the analyte. In the first case, increasing
signal with increasing value of the analyte, and a larger signal in the standard than in the unknown
relates to a smaller value of the measurand in the unknown. For signals showing the negative behaviour
(smaller signals with increasing value of the measurand), a smaller signal in the unknown relative to the
signal from the standard points to a higher value of the measurand in the unknown sample.
This dependence of the signal on the value of the measurand can be utilized for the quantification of this
analyte. If the signal depends also on other concomitant species these are regarded as interferents and, if
not properly dealt with, this (secondary) dependence can lead to inaccurate measurements.
Similarly, this type of comparison is also useful for identification of chemical moieties. For the
identification of a species, one frequently resorts to spectroscopic measurements, where we measure a
spectrum as a function of wavelength (or wavenumber). The matter of interest in these cases is not only
the size of the signal but also the wavelength axis. To point out a certain wavelength, e.g., 280 nm, along
the wavelength axis it is useful to have a standard with one or more distinct features around this
wavelength. If a feature expected at 250 nm shows up at the (uncalibrated) nominal wavelength of 275
nm, then the wavelength scale can be adjusted (calibrated) by a factor of 1.1 to give the true physical
wavelength.
This type of calibration is automated and most of the time it is not noticed by theoperator in many
modern instruments so that the reliability of the operation is increased.
Such a comparison of a wavelength scale is also required for electrical measurements (potential,
current), but this calibration can often be taken for granted if the measuring instruments (e.g., voltmeter)
are properly attended. In liquid chromatography there is the need to calibrate the time axis in terms of
the eluent stream (mL/min) in order to facilitate the correlation between elution times (elution volumes)
and chemical substances.
Here we concern ourselves mainly with quantitative measurements of chemical species and the role of
calibration in this process. One calibrates the reading of a measurement system in order to give it a
quantitative interpretation. Another type of comparison will be dealt with later (library search, pattern
recognition) where the aim is to recover qualitative information regarding the nature of a chemical
species.
Quality of calibration:
No quantitative measurement can therefore be better than the intrinsic quality of this comparison process
called calibration, and there are different ways to improve this comparison process as shall be seen later.
One factor is the repeatability of the measurement itself.
Suppose the measurement gives highly variable readings, it will then tend to do so both for the standard
and the unknown. The signal size is therefore not properly defined leading to poor predictions of the
(value of the) measurand. In practice the variability for the standards may be different (and often
smaller) than the variability of the measurement of the sample, thus the latter is limiting the quality of
calibration.
The other factor correlates to the value of the measurand in the standard. Although ideally this should be
known with high accuracy, there is always some uncertainty remaining since its value is also determined
by measurements, albeit of higher quality than routinely obtainable. Even with highly reproducible
measurements this uncertainty regarding the value of the standard leads to signal imperfections in the
comparison process.
The value given to a standard usually carries a smaller (relative) uncertainty if the standard is produced
from pure chemicals with well-defined stoichiometry. It has a greater uncertainty if the standard is a
reference material with a matrix similar to the samples. This is not be the disadvantage as the calibration
is then particularly relevant to the problem and one needs to worry less about matrix effects.
The final factor limiting the quality of calibration, and in many analytical systems the overwhelming
one, is the validity of comparison itself. In calibrating an instrument, one must be reasonably sure that it
should respond to the measurand in the standard in the same way it does to the measurand in a different
environment, the unknown. Physical differences between sample and standard also frequently hamper
the calibration by invalidating the comparison process.
This problem is considerably more often to occur if calibration is based on measurements of standards
prepared from chemicals of high purity, and you can expect the advantages from the use of in-type
standard reference materials for calibration, if available. Which particular physical properties can affect
the resultsand need to be identical (or at least very similar) in standards and unknowns depends on the
principle of the measurement and the physical state of the sample.
Among them are temperature, pressure, viscosity, turbidity, particle size, surface roughness, and
thickness. Differences in response can also be caused by impurity and the measurement is then said to be
of insufficient selectivity. Differences could also be traced to impurity or loss whereby a higher or lower
signal might be obtained.
One of the most difficult aspects of analytical work is the judgement regarding the validity of a
comparison and can ultimately only be done by checking the accuracy of results on reference materials.
Alternatively, results obtained on several samples by two procedures are compared that is based on a
completely different measuring principle can show the accuracy of the data. For instance, it might be
possible to assess the accuracy of a spectrophotometric determination of phosphate by one based on ion
chromatography.
Frequency of calibration and recalibration:
Although great efforts are being made to produce analytical instruments that give stable readings
("response") over time, the long-term stability is usually limited. This is partly the reason for the
importance of calibration in practice: while for an analytical balance it might be sufficient to calibrate it
once every two months (weighing serves to determine a physical measurand, mass). Most chemical
measurements are calibrated daily or even more frequently than that. In this way chemists attempt to
reduce the day-to-day (and even sample-to-sample) variability of their measurements as much as
possible. Thus, on each working day every chemist in the world produces his/her own scale that mayor
may not be significantly different from all other ones.
Absolute vs. relative methods of analysis
Measurements that are evaluated by comparison with those made on standards are called relative
methods. Absolute methods can be evaluated without comparative measurements, but they rely instead
on fundamental physical and chemical constants and principles.
Certain electrochemical measurements rely on a definite oxidation state of a chemical substance before
and after the measurement, and therefore the transferred charge is directly convertible to the amount to
substance (in mol), the conversion factor being Faraday's constant. Similarly, volumetric measurements
(e.g., titrations) rely on the presence of welldefined chemical species, before and afterwards, that are
related to each other by stoichiometric coefficients from a reaction equation. The knowledge of the
particular reaction equation and of atomic masses is therefore sufficient for the quantitation of the
measurand.
In these (and similar) instances, no calibration for the actual measuring step is necessary provided the
titrant is available in high purity. If, however, other operations are also involved in the analytical
procedure, e.g., decomposition, extraction, or clean-up, it may be advisable to carry a standard through
the entire procedure in order to check for contamination, losses, and degradation of the measurand.
This is why in practice very few procedures are operated completelywithout standards. Another
complication for such absolute methods is the fact that the actual measurement is done in terms of
volume (of the titrant), the exact concentration of which is rarely known with sufficient accuracy.
Therefore, in spite of the validity of the fundamental stoichiometric relation, the actual volume is
calibrated by comparison with the result of the measurement of a standard.
An example is the titration of a strong acid with a strong base, e.g., NaOH. NaOH solution is made up to
have a molarity of 1.0 mol/L. Instead of relying on the dilution process, a measurement is done on a very
well-defined amount of strong acid, e.g., HNO3. If 0.0100 mol HNO3 is taken, one expects a volumetric
consumption of NaOH solution of 10.0mL; instead, 10.3 mL are consumed, and the relation between the
volume of titrant and the amount of acid is adjusted by the factor 1.03.
This proportional adjustment corresponds to the construction of a separate calibration function for this
particular batch of titrant using a single standard. It is necessary only because in practice the volume of
the basic solution is measured instead of the amount of base, and so does not invalidate the fundamental
stoichiometric equation underlying this volumetric measurement.
The jargon "standard-less" procedure also often relates to relative methods that do not require calibration
by the operator in the laboratory or in the field. Generally, these procedures are characterized by a stable
relationship between the sample and the instrumental response leading to great intervals between
calibrations. In the extreme, the calibration might be valid for the lifetime of an instrument and therefore
49
carried out prior to taking in the factory. If this is possible it is a great convenience for the analyst, but
the responsibility for adequate calibration rests still with him/her. It is therefore wise to check in regular
interval whether the system still operates according to the same response characteristics. The sequence
of experimental steps in calibration are summarized.

Obtain (or prepare) standards

Measure standards

Derive calibration model

Measure unknowns

Apply calibration model to data from unknown

Recalibrate regularly
General flowchart of calibration
Calibration protocols and calibration models
The simplest calibration is one that use only one standard. One may be forced into such a protocol when
only one sufficiently reliable standard is available. In this instance, it is supposed that for a zero
measurand (e.g., zero wavelength, retention time, mass, amount, or concentration) the analytical system
gives zero (or a very well-known value other than zero) response (signal, reading). Betweenthese two
values at intermediate mass of the analyte, one resorts to a linear interpolation, but no extrapolation
beyond the value of the standard is recommended. The corresponding model for measuring
concentration is
Signal = constant × concentration
y = b1 c

50
The constant b1 is called the sensitivity and can be regarded as a proportionality constant between the
signal and the concentration, or more generally, between the signal and the analyte. This proportionality
is usually valid over a restricted range of values of the measurand; below this range the signal is too
small to beuse, and above there are generally deviations from this proportionality, so that the value of b1
is no longer the same.
The region of validity of b1 is called the working range of an analytical system; it can often be extended
to higher values of the analyte by suitable dilutions, while for reaching lower values, more involved
chemical operations may be necessary, e.g., pre-concentration of the analyte. At higher values of the
measurand the curve tends often to bend towards the x-axis.
As long as there is a noticeable change in signal as the concentration changes, one operates within the
dynamic range of the method although the sensitivity decreases continuously for such a bent curve.
If the response at zero concentration is not known beforehand it is measured in the course of calibration.
A measurable signal at zero (or negligible) concentration of analyte is either due to contamination of the
sample or due to inadequate recovery of the net signal from the background, or both. The simplest
model for this two-point calibration that can also be extended to multiple points is:
signal = blank + constant x concentration (or amount)
y = b0 + b1 c
The term b0 thus indicates the magnitude of the blank whereas b, is again a measure of sensitivity. More
formally, a test for statistical significance will reveal the necessity to include the b0 term in the
calculations.
For a curvilinear dependence of signals from measurand, other models must be employed, such as the
quadratic model:
signal = blank + (constant1 x concentration) +(constant2 x concentration2)
Y = b0 + (b1 c) + (b11 c2 )
The more involved the model the greater the required number of standards. Another reason for using a
larger number of standards is in multivariate calibration when more than one analyte is considered
simultaneously, or if - due to the lack of selectivity - corrections of the signal for the contributions of the
concomitant species are applied.
This not only requires the use of multiple standards, but also the knowledge of the concentrations of all
species contributing to the signal in any of the standards. In mathematical terms the simplest model
involving k species corresponds to:
y = b0 + b1c1+ b2c2+ b3c3+ …. +bkck
but much more involved ones (e.g., hyperbolic or sigmoid) are also in use. For instance, in Xray
fluorescence spectrometry it is not uncommon to measure 30- 60 standards. A good strategy in
calibration, however, is to use only as parsimonious a model as possible and to test for the most
parsimonious model consistent with the data by a formal statistical testing procedure, thereby avoiding
any overfitting of the experimental results.

51
Once the calibration models are derived by one of the methods presented, they are employed to predict
the measurand in a sample other than a standard. In order to do so, the models need to be reformulatedin
terms of their inverse. These inverse functions are called analytical models. Such an analytical model for
Eq. is:
c = Y/b1
For Eq. 2 the analytical model is:
c = (y – b0)/b1
Analytical models for the more complicated calibration models are either obtained by algebraic methods
or - equally important – by numerical approaches in an iterative manner.
Calibration modes and protocols:
While calibration is generally practiced in analytical chemistry the exact mode ofoperation varies
depending on the analytical method. In some methods it isenough to measure a single standard; this is
called single-point calibration. Sometimes a two-point calibration is practiced, one is a standard at a low
concentration (or amount) value, another is at a high concentration (or amount) value. The two are
chosen to bracket the values of the unknowns; the lower onemight be the blank value.
These protocols are all versions of a calibration mode that is called external calibration because the
samples containing the unknown amount, and the standards containing the known amount are separate
throughout. In addition to the external calibration some other modes are also important: internal
calibration, standard additions method, and isotope dilution techniques.
If prior to the determination, a fixed amount of a different, but similar substanceis added to the sample,
this substance is called the internal standard and themode of operation is called internal calibration. The
addition serves the purpose ofcontrolling a critical step that would otherwise introduce a large element
of uncertainty. Therefore, it is wise to add the surrogate substance as early as possible inthe analytical
procedure.
Care has to be taken, however, that the added substance is well mixed with the sample and is in a
physical state (particle size. surface, etc.) and binding state comparable with that of the measurand. The
assumption in the internal standard mode is that the measurand shows the same behaviour in all the
critical steps; thus, the ratio between the data of the measurand and the surrogate substance constitutes
more reliable information than the data of the measurand itself.
A related mode is the method of standard additions since one uses the measuranditself for internal
calibration. In order to do so, the sample is split into severalsubsamples before the analysis. One
subsample is treated as usual, while one addsincreasing amounts of the measurand to the other
subsamples.
The purpose of this mode is generally to correct for proportional errors that stem from the differences in
response between standardsin clean solution (e.g., water or ethanol) and response in the matrix loaded
sample. In effect one attempts to correct for different slopes between the two curves. Inorder to have a
good estimate of the sensitivity it is necessary to add enough spikeso that the signal is at least doubled.
As one frequently prepares two or threespiked samples this leads to multiple work on single samples,
this being an obviousdisadvantage in terms of throughput.

52
The measurement on the unknown is evaluated by:
c = b0 – bl / b1
wherebl is the independently determined blank signal that applies to the particular measurement. It is
equally important to be alert to the assumptions that need to be fulfilled if this mode should lead to
success. One of them is that the curve has to be strictly linear even in the lower range, where, due to the
presence of the measurand in the sample, one cannot produce data, but extrapolates to zero. Another
crucial assumption, particularly in trace analysis, is that there is no appreciable blank in the sample for it
is impossible to test this fact experimentally.
The most advanced form of internal standardization is the isotope dilution mode. Here one opts for a
spike that is, chemically speaking, most similar to themeasurand, yet still discernible from the original
one. It is the identical chemical substance, but with at least one atom in the structure replaced by another
isotope.
Frequently one replaces a hydrogen by a deuterium atom or a 12C by a 13C in an organic substance. If
only ions or atoms are determined, then one adds a known amount of the ion/atom in different isotopic
abundances so that the isotopic ratio of the sample is altered: the more of the measurand originally
present, then the smaller the resulting overall change.
For this technique an isotopically selective detector is required, so that in practice the detection is
generally done by mass spectrometry. The technique is then termed isotope dilution mass spectrometry
to highlight the essential feature of this operation.
In order to give the best accuracy some prerequisites are necessary:
(a) The isotopically discernible spike needs to be added in a similar amount to that present in the sample.
This generally requires a rough pre-analysis.
(b) The spike must be in the same chemical state as the measurand is in the sample. For example, if the
substance is Cr(H20)6 3+, then the spike must also be the hexahydrate of Cr(III), and no Cr04- , or some
other form of chromium.
(c) One must be certain that prior to any chemical pre-treatment of the sample thespike is
homogeneously mixed with the native measurand.
However, if viable, this technique corrects for very incomplete recoveries thatmoreover may vary from
sample to sample. Care must be taken not to contaminatethe sample after addition of the spike.
Calibration of instruments
Calibration of Spectrophotometer:
Wavelength calibration of a spectrophotometer can be checked by using a holmium oxide filter, as a
wavelength standard. Holmium oxide glass has a number of sharp absorption bands, which occur at
precisely known wavelengths in the visible and ultraviolet regions of the spectrum. Holmium oxide
filter wavelength peaks are given below:
Ultraviolet range with deuterium lamp: 279.3 nm, 287.6 nm
Visible range with tungsten lamp: 360.8 nm, 418. 5 nm, 453.4 nm,536.4 nm, 637.5 nm

53
 In double-beam spectrophotometers, zero is adjusted with sample and reference beam. Then
holmium oxide filter is placed in sample beam. The wavelength control is manually scanned through
each wavelength, until the absorption peak is found, always approaching each point from the longer
to the shorter wavelength.
The spectrum is then recorded. The wavelength calibration can also be checked in the visible region
by plotting the absorption spectrum of a didymium glass, which has been in turn, calibrated at
National Bureau of Standards, USA.
Calibration of IR Spectrophotometer
In order to check IR spectrophotometer, spectrum of a polystyrene film is run and the absorption
spectrum is compared with that provided by the company. In case any difference in the absorption
spectrum is detected, the instrument has to be thoroughly checked and recalibrated. To facilitate speedy
recalibration of low resolution FIR instruments, special broad-band wavelength calibrators are often
used.
The transmittance of the filter obtained between 800 and 350 cm-1 on a Perkin-Elmer 457
spectrophotometer shows that it provides three accurate calibration points: one due to the polythene
matrix itself (at 722 ± 1 cm-1 ) and the other two (at 590 ± 2 cm-1 and 500 ± 1 cm-1 ) due to mercuric
oxide.
Calibration of Flame Photometer
Calibration Curve Method
In order to determine the concentration of an unknown element X in a given sample, or a series of
samples, the instrument is first standardised by making a calibration curve. For this, it is desirable to
know the probable range of concentration of Xin the sample, so that a series of calibration standards can
be made up; having a suitable spread over the range expected. The instrument is first adjusted to give a
zero-metre reading, when pure distilled water is aspirated into the flame.
The instrument settings are then added to give the full-scale reading, with the most concentrated
standard sample. Without altering this adjustment, emission intensities for the remaining standards and
the unknown are measured. Calibration curves are thus plotted and the concentrations read off against
this calibration, concentration of the unknown is obtained from the calibrated curve. Since it is difficult
to obtain a flame of high long-term stability, it is usual to check the calibration at frequent intervals by
spraying a standard. This ensures that drift can be measured and taken into account. In fact, calibration
curves are difficult to reproduce on the same day; virtually impossible on different days.
Internal Standard Method
The flame system is a critical part of the flame photometer and its long-term stability is dependent upon
(i) fuel flow, (ii) air pressure, (iii) nebulisation rate and (iv) burner/atomising chamber temperature.
While the first two factors can be controlled effectively to within acceptable limits by suitable
instrument design, the other two are less easily controlled in this way. To overcome variation from these
sources, internal standard procedure has been developed.
In this method a fixed quantity of the internal standard element is added to the blank sample and
standard solution. The radiation of the element is measured together with those of the elements under
analysis by dual detectors, or by scanning successively the two emission lines. The ratio of the emission
54
intensity of the analysis line to that of the internal standard line is plotted against the concentration of the
analysis element on log-log paper, to prepare the calibration curve for a number of standards. As a ratio
rather than an absolute light intensity that is being measured, because of which fluctuations due to
variable operation of the nebuliser-burner system, are greatly minimised. This procedure also reduces
errors due to differences in the composition of the test and standard solutions.
For analysis of alkali metals and even of other elements, lithium has been preferred as an internal
standard, as it satisfies most of the above-mentioned requirements which overcomes the above
problems. The junction consists of several small pores in the actual body of the electrode which can be
wiped clean each time the electrode is rinsed. There is direct contact between the sample and the gel,
thus resulting in faster readings. Additionally, this allows the electrode to be stored dry and eliminates
the need for a special storage solution. The gel-filled electrodes operate in a wide temperature range (-5
to 100°C) and are available in various shapes, sizes and lengths to meet varied requirements.
The electrodes should never be stored dry. The preferred choice for storing is KCl solution of any
concentration between 2.0M and 3.8 M. Another good choice is buffer solution (pH4). If it is necessary
to clean the probe with acid, caustic, solvent or other cleaning solution, it is best to soak the electrode in
KCI solution after cleaning and prior to use or calibration. This will re-condition the bulb and reference,
extending the probe life and improving calibration accuracy.
Calibration of pH meter
Buffer Solutions A buffer may be defined as a solution whose pH remains nearly constant, despite the
addition of a substantial quantity of acid or base. Buffers are employed for the standardisation of pH
cells. The cell emf is measured for given buffers and related to the known pH values for the buffers. The
pH value of an unknown solution is then derived from this calibration. Many buffer solutions have been
reported in literature. There are also British standards and standards of NBS on buffers.
Buffer tablets are commercially available, which when dissolved in an appropriate quantity of fresh
distilled water, give buffer solutions. These tablets may contain the buffer material admixed with
substance, which aid in tabletting but do not significantly affect the pH. Buffer tablets for pH 4,7 and 9.2
are available commercially.
Buffers should be protected from exposure to the atmosphere, where gases such as carbon dioxide,
ammonia and oxygen are present, as they will tend to change the pH value of the buffer solution. Since
air would enter the bottle everything the buffer containing bottle is opened, a buffer solution would have
a limited useful lifetime, when reliable pH measurements are required. Also, the pH value of a buffer
solution depends upon the temperature and the actual value for the testtemperature can be found on the
label on the bottle.
--------------------------------*-----------------------------------

55
 CHAPTER 7
STIOCHIOMETRIC CALCULATIONS
Analytical chemistry is concerned with the measurement of concentration of analytes, from which their
respective masses can be easily calculated. Thus, solutions of known concentrations are prepared and are
employed for calibration of instruments or for titration of sample solutions. We can calculate the analyte
mass from its concentration and volume. All the calculations involved in obtaining concentration of
analytes require a deep knowledge of stoichiometry. In the present section fundamental concepts and the
use of stoichiometry in titrimetric analysis to determine mass of analyte have been reviewed.
Some fundamental concepts:
We know that an element consist of atom and the weight of the atom represents its atomic weight. A
gram-atomic weight of an element has equivalent number of atoms of that particular element as there are
carbon atoms in 12 g of carbon 12. This number is known as Avogadro's number and has a value of
about 6.022 X 1023 atoms/g-at wt. In nature, elements consisting of mixtures of isotopes are found.
Their atomic weights are therefore an average of weights of the isotopes of every element, taking this
into consideration the virtual natural abundance of the element. In simple when we add the atomic
weight of the element and the total sum is called the gram molecular weight.
Dalton
For reporting masses of large proteinaceous compounds such as mitochondria, ribosomes,
chromosomes and viruses the unit Dalton is used. Mass of carbon-12 atom is 12 daltons, and 1 dalton is
therefore 1.661 X10-24 g . With respect to avogardo principle molecular weight is equal to total number
of Daltons in the molecule.
Moles
Even though we know, that a reaction is possible only with definite ratios of reactants then also the
analyst is unable to calculate number of molecules participating in the reaction. Since the relative masses
of the reacting species are known the reactions can be described with ease on the basis of relative masses
of reactants. The concept of mole has been developed to simplify the calculations. A mole which is
Avogadro's number of ions, atoms, molecules or other species, can also be expressed as atomic,
molecular, or formula weight of anycompoundin grams. Moles of any compound can be calculated.

 Moles = grams / formula weight (g/ mol)

In above equation formula weight represents the molecular weight of that particular compound.

Expressing concentrations of solutions

Molarity

The concept of mole is useful to express concentrations of solutions, where it is necessary to know the
ratios of reactants (volume of different reactants) participating in the reaction. Solution of any substance
is said to be one molar if the one litre of the solution contains one mole of that substance. Molar is
abbreviated as M and molarity of any solution is expressed in moles per litre.
56
Normality

A solution containing one equivalent per litre is said to be one normal. An equivalent is the mass of
substance which provides Avogadro's number of reacting units. Reacting unit is an electron or a proton.
Number of equivalents is given by

 Number of Equivalents (eq) = Wt (g) / Eq wt ( g/ eq) = normality (eqL) × Volume (L)

Equivalent weight is that weight of a substance in grams that will furnishone mole of the reacting unit.
Thus, for HCl, equivalent weight is equal to the formula weight. Equivalent weight is dependent on
chemical reaction. As with molarity, an analyst usually works with milli-equivalent amounts, and

 meq = mg / eq wt (mg/meq) = normality (meq/m L) × m L

Formality

An analyst sometimes uses the term formality for ionic salt solutions not existing as molecules in solid
or in solution. Operationally, formality is identical to molarity.

Molality

A solution is said to be one molal if it contains one mole perkilogram of solvent. Molalconcentration is
very usefulwhile measuring properties such as lowering of vapour pressure, freezing point depression
and osmotic pressure. Molal concentrations are independent of temperature.

Density Calculations

Concentrations of many concentrated acids and bases are generallyreported in terms of percent by
weight. So it becomes essential for an analyst to maketheir molar solutions accurately. For making their
molar solutions density of that substance should be known. Density is defined as weight per unit volume
at a given temperature. Sometimes specific gravity instead of the density is given. When specific gravity
is referred to water, density is given by the following formula.

specific gravity X 0.99821

Analytical and equilibrium concentrations Analysts make solutions of known concentrations but the
partial or total dissociation of dissolved substancesgives equilibrium concentrations of various species.
Acetic acid dissociates into proton and the acetate ion. Analytical molarity is given by Cx, whereas
molarity at equilibrium is given by the notation [X].

Dilutions Dilute solutions should be prepared from stock solutions whose concentration is high.
Millimoles of concentrated stock solution which is employed for dilution purpose will be equal to
millimolespresent in final diluted solution.

Expressing results obtained

57
Results can begiven as concentration, on basis of weight or volume. Units employed for substance under
investigation will vary. Gram (g) is the widely used unit in many analysis involving large amounts of
analyte. For constituents present in small quantities, analysts use smaller units such as milligram,
microgramand nanogram. The most popular unit of volume is liter (L). Milliliter is employedgenerally
in titrimetry.

Solid samples

To calculate percent on a weight/weight basisfollowing formula can be used,

% (wt /wt) = [wt solute (g) / wt sample (g)] X 102 (%/g solute /g sample)

Trance concentrations are generally reported in smaller units.

pt (wt/wt) = [wt solute(g)/wt sample(g)] X 103 (ppt/g solute/g samples)

ppm (wt/wt) = [wt solute(g)/wt sample(g)] X 106(ppm/g solute/g samples)

ppb(wt/wt) = [wt solute(g)/wt sample(g)] X 109(ppb/g solute/g samples)

table: Units used to express trace constitutents

Unit Abbrevation wt/wt wt/ vol vol/vol

Parts per million ppm mg/kg mg/L µL/L
(1 ppm = 10-4%) µg/g µg/mL n L/mL
Parts per billion ppb µg/kg µg/L n L/ L
1ppb = 10-7 % = 10 -3 ppm ng/g ng/mL p L/mL
Milligram percentage mg % mg/100g mg/100mL -

Liquid Samples

Results for liquid samples can be reported either on weight/weight or weight/volume basis. Some of the
calculations used for reporting results are given below:

% (wt/vol) = wt solute (g)/ vol sample (m L) X 102 (%/g solute/m L sample)

ppm (wt/ vol) = [wt solute(g)/vol sample(m L)] X 106 (ppm/g solute/m L samples)

ppt (wt/vol) = [wt solute(g)/vol sample(m L)] X 1012(ppm/g solute/g samples)

ppb(wt/wt) = [wt solute(g)/vol sample(m L)] X 109(ppb/g solute/g samples)

58
The unitsweight/weight and weight/volume are related through density. They are almostequal for dilute
aqueous solutions having a density of 1 g/mL. If a liquid analyte is in solution withan another liquid,
results are reported on a volume/volume basis. Results of a gas analyses may be given on a
weight/volume, weight/weight, or volume/volume basis. Analysts mostly employmilliequivalent (meq)
unit rather than weight to expressthe quantity of different electrolytes in biological fluids.
Milliequivalent, is number of millimoles of a substance under investigation multiplied by charge on the
ion of that substance. Results are usuallyexpressed as meq/L. Milliequivalents of cations are equal to
milliequivalentsof anions. Milliequivalents of ananalyte can be calculated by the following formula:

meq = mg/eq wt (mg/ meq) = mg/ f wt (mg/ mmol)/n meq/mmol)

n = charge on ion

Reporting concentrations

It is not always necessary to determine the substance in the formit exists or for which the analyst wants
to report the obtained results. In case of iron present in an ore, for example, iron in form of Fe2O3can be
determined and then reportedas % Fe. Or ironin form of Fe2+can be determined and reported as %
Fe2O3.

Weight criteria employed forexpression of results with solids or biological tissues have also been
mentioned. The sample can be weighedin three forms; wet, dry, or ashed. This mayapply to fluids as
well, although volume of fluid is generally used. If the analyte is sensitive to heat, itshould not be
heated.

Volumetric analysis

Volumetric analysis iswidely used technique to obtain reliable results, especially for smallquantities of
substance. The analysis is less time consuming andcan also be combined with otheranalytical techniques
for detectionof completion oftitration reaction. Titration process can also be automated. Titrations can
also be performed manually where high accuracy is required for small number of samples. Automated
titrations are helpful when titration of large number of samples is required. Here, the types of titrations
aredescribed in addition to the principles which are applicable to different types of titrations.

Technique of Titration

59
In the process of titration, an analyte reacts with a reagent, of known concentration, which is addedfrom
a burette dropwise. The reagent filled in the burette is knownas titrant or standard solution. The volume
of reagentwhich is added from the burette to complete the reaction with the analyte is measured. As the
concentration and reaction between analyte and titrant is known, the quantity of analyte can be easily
calculated.

Noticeable change in some property of solution should be observed on the completion of the reaction. A
change in color physical or electrical properties usually seen. When titration of acetic acid is done with
sodium hydroxide, pH of the solution increaseson the completion of the reaction. Anindicator brings
about a change in color of the solution when reaction is just complete. Equivalence point is the point at
which stoichiometric amount of titrant has been added. At the end point the reaction is completed.

Standard solutions

A standard solution is a solution containingfixedamount of asubstance of high purityknown as primary

standard and its dilution to a known volume. In case, if the substance contains some number of
impurities, a solution is prepared to get the desired concentration andthis is standardized by titration of a
fixedamount of a primary standard. Forexample, NaOHis hygroscopic in nature and may absorb
appreciable amount of water vapour from the atmosphere and therefore its standard solution can’t be
prepareddirectly. NaOH is first standardized by potassium acid phthalate (a primary standard).

Requirements for a primary standard are:

a. Primary standard should be of high purity. Limit ofimpurities present should be very less and only
0.01% to 0.02% is acceptable if the impurities are accurately known.

b. Primary standardmust be stable atroom temperature and temperatures used for drying. A primary
standard is dried before taking its weight.

c. Primary standard should be easily available and should not be expensive.

d. Primary standard should have high formula weight. Due to this a large quantity of it will be required
for titration purpose and the error in weighing will be reduced.

There are four types of titrations employed in the analytical laboratory:

1. Acid-base titration: A wide variety of substances, inorganic as well as organic, are either bases
or acids and their titration can be easily done with the help of a standard solution of a strong acid

60
 or a strongbase. The end points can be easily detected in this type of titration, either by using a
solution of an indicator or by employing a pH meter to notice the change in pH of the solution.
By performing titration in a non-aqueous solvent, the acidityand basicity of various organic acids
and bases can be increased. A sharp end point is obtained as a result of this type of solvent and
this method is most suitable for titration of weaker acidsand bases.
2. Precipitation titration: In thistype of titration, an insoluble productis formed by the reaction of
the titrant which is added from the burette with the analyte present in the flask. A precipitate of
silver chloride is formed when titration of chloride ion is done with solution of silver nitrate.
Indicators can also beemployed here for detection of the end point, or potential of solution can be
examinedelectrically.
3. Complexometric titration: In this type of titration, the titrant forms a complex with ametal ion
(analyte). The complex formed is water soluble. A chelating agent is mostly used as the titrant.
Ethylenediaminetetraacetic acid (EDTA) is the most widely used chelating agents. EDTA
reactseasily with many elements, and the reactions can be controlled without any difficulties by
adjustingpH. Indicators can be employed forformation of a colored complex with the analyte
(metal ion).
4. Redox titration: In these types of titrations, titration of an oxidizingagent with a reducing agent,
or vice versa is done. In a reaction between an oxidizing agent and a reducing agent the oxidizing
agent gainselectrons whilethe reducing agentloses its electrons. To get a sharp end point there
should be enough difference between oxidizing and reducingabilities of these Analytical
Chemistry / Instrumentation Fundamentals of Analytical Chemistry Stoichiometric Calculations
agents. Appropriate indicators can be used forthese type of titrations, or the end point can be
detected electrically.

Calculations generally used in titration

Some fundamental concepts are given below:

Moles = g/ f wt (g/mol)

Milli moles = mg/ f wt (mg/mmol)

Molar concentration = M = mol/ L

M = mmol/ m L

61
By rearrangement of these equations, expressions for calculation of other quantities can be
obatained.

M ( mol/ L) X L = mol

M (mmol/m L) X m L = mmol

g = mol X f wt (g/ mol)

mg = mmol X f wt (mg/mmol)

g = M (mol/L) X L X f wt (g/mol)

mg = M (mmol/m L) X m L X f wt (mg /mmol)

Percentage of analyte A reacting on a 1:1 mole basis withtitrating reagent can be easily calculated
with the help of the formula given below.

% A = fraction analyte X 100% = mganalyte/ mgsample X 100%

Mmol X f wt analyte (mg/ mmol) / mg sample X 100%

Mtitrant (mmol/m L) X m L titrant X f wt analyte (mg/mmol)/mg sample X 100%

In many reactions, the compoundsdon’t react on a 1:1 mole basis and so the above calculationcannot
be applied to all the reactions. However, a generalized formula based on balanced equation for
reactions can be written which will be applicable to all reactions. For a general reaction.

a A X t T -> t T -> P

where A is ana analyte, T is titrating reagent and analyte and titrant react in ratio a/t to form
products P, then formula for molarity calculation is:

mmol A = mmol T X a /t mmol A / mmol T)

mmolA = MT (mmol/m L) X m LT X a/t (mmol A / mmol T)

mgA = mmolA X f wt (mg/ mmol)

mg A = MT (mmol/m L) X m LT X a/t (mmol/ mmol T) X f wt (mg/mmol)

Some steps are listed below to obtain a general expression to calculate percent analyte A in a sample.

62
% A = fraction anylate X 100% = 100% = mganalyte / mg sample X 100%

Mmoltitrant X (a/t) (mmolanalyte / mmoltitrant) X f wt analyte (mg/mmol)/mg sample

Standardization and titrant calculations:

When a highly pure titrant is not available, then the concentrationof approximately prepared
titratingreagent must be known bystandardizing the titrant with a suitable primary standard. From the
volume of titrating reagentemployed for titration ofthe primary standard, the molar concentration of
the titrating reagent can be calculated. Assuming the analyte A in equation to be the primary
standard, or, combining all steps:

mmolstandard = mgstandard / f wt standard (mg/mmol)

mmoltitrant = Mtitrant (mmol/m L) X m L titrant

mmolstandard X t/a (mmoltitrant/ mmolstandard)

Mtitrant (mmol/m L) = mmolstandard X t/a (mmoltitrant/ mmolstandard)

Mtitrant (mmol/m L) = mgstandard X f wt standard (mg/mmol) X t/a (mmoltitrant/ mmolstandard)/m L titarnt

Mtitrant (mmol/m L) = mgstandard X f wt standard (mg/mmol) X t/a (mmoltitrant/ mmolstandard)/m L titarnt

Reaction of the analyte and titrant in different ratios

Various compounds can react to form different products. Factor employed in calculation
ofmillimolesof such a compound from the millimoles of titrating reagentwhich react with it
dependsuponthe particular reaction. For example, sodium carbonate can either react as a diprotic
base or amonoprotic base.

Back titration

When the reaction is very slow and a sharp end point is difficult to obtain then a back titration is
performed. In titrating antacid tablets with a strong acid back titration is very useful. In these
situations, a back-titration furnishesreliable and accurate results. In this titration a weighedquantity
of the reagent (normally the titrant) is added to the sample solution so that the titrant is in excess in
the solution. After the reaction is complete, the quantity of excess (unreacted) reagent is determined
by titrating it with another standard solution; the reaction of the analyte may also be increased
byexcess reagent present in the solution. By knowing number of millimoles of reagent added and by
63
 measurement of number of millimoleswhich remain unreacted, number of millimoles of sample
reacting with reagent can be calculated by the following formula:

Mmol reagent reacted = mmol taken – mmol back – titrated

Mg analyte = mmol reagent reacted × factor (mmol analyte/ mmol reagent) × f wt analyte
(mg/mmol)

Reacting units in normality calculations

Acid-Base:

The reacting unit for acids and bases is proton H+. If the compound reacts as an acid, number ofreactive
protons it possesses per molecule should be determined. If it reacts as a base, number of protons it will
react with per molecule must be determined. Then,

Eq wt = f wt / no. of H+

Reduction-Oxidation:

Reacting unit in reduction-oxidation reactionis electron. A reducing agent loses electrons and is thereby
oxidized andan oxidizing agent gains electrons and is thereby reduced. Thus,

Eq wt = f wt/ no. of moles of electrons gained or lost

Titer

For volumetric analysis, titer of the titrating reagent is usually calculated. Titer is weight of analyte
which is chemically equal to 1 mL of titrant. Titer isgenerally expressed in milligrams.

----------------*--------------------

64
 CHAPTER 8

TITRIMETRIC METHODS OF ANALYSIS

INTRODUCTION

Titrimetric or volumetric analysis is one of most important technique of analytical chemistry and its
calculations involved are based on simple stoichiometric relationships. The term titration refers to
process of measuring the volume of titrant required to reach equivalence point. However, from a
rigorous standpoint the term titrimetric is preferable because volume measurements need not be
confined to titrations.

TYPES OF TITRATIONS

Chemical reactions which may serve as the basis for titrimetric determinations are of four types:

 Acid –base

If HA represents the acid to be determined and B the base, the reactions are

HA + OH-  A - + H2O

The titrants are generally standard solutions of strong electrolytes, such as sodium hydroxide and
hydrochloric acid.

 Oxidation-reduction (redox) Chemical reactions involving oxidation-reduction are widely used

in titrimetric analyses. For example, iron in the +2-oxidation state can be titrated with a standard
solution of cerium (IV) sulfate:

Fe2+ + Ce 4+ Fe3+ + Ce3+

 Precipitation The precipitation of silver cation with the halogen anions is a widely used
titrimetric procedure. The reaction is

Ag+ + X- ----> AgX

 Complex formation An example of a reaction in which a stable complex is formed is that

between silver and cyanide ions:

65
 Ag++ 2CN- ----- Ag(CN)2 –

This reaction is the basis of the so-called Liebig method for the determination of cyanide. Certain
organic reagents, such as ethylene di amine tetra acetic acid (EDTA), form stable complexes with a
number of metal ions and are widely used for the titrimetric determination of these metals.

Conditions for Reactions Used in Titrimetric Analysis

A reaction must satisfy following requirements:

1. It must proceed as per a definite chemical equation. There should be no side reactions.

2. The reaction should be almost complete at the equivalence point.

3. An indicator should be available, or some instrumental method may be used to tell the analyst to
stop the addition of titrant.

4. It is desirable that the reaction should not be time consuming, so that the titration can be
completed in a few minutes. Consider an example of a reaction well suited for titrations.

The reaction goes to virtual completion. At the equivalence point the large change in pH of the
solution takes palce by a few drops of titrant, and a number of indicators are available which respond
to this pH change by changing colour.

CONCENTRATION SYSTEMS

 The methods used by the analytical chemistry to express the concentration of a solution, viz.,
relative amounts of solute and solvent.
 The systems of molarity and normality are most commonly used since they are based on the
volume of solution, the quantity of analyte measured in a titration.
 Formality and analytical concentration are useful in situations where dissociation or complex
formation occurs.
 The percent -by -weight system is commonly usedto express approximate concentrations of
laboratory reagents.
 For very dilute solutions parts per million or parts per billion units are convenient.

MOLECULAR AND FORMULA WEIGHTS

66
 The mole (or mol) is defined as the amount of a substance which contains as many entities.
The entities may be atoms, molecules, ions, or electrons.
If particles are molecules, weight in grams of a mole of substance is called the gram - molecular
weight (usually molecular weight).
If the particles are atoms, the weight in grams of 1 mol of the substance is called the gram
atomic weight.
The term gram formula weight (or formula weight) is the addition of the atomic weights of all
the atoms in the chemical formula of a substance and is normally the same as the molecular
weight.
We shall use the term molecular weight as synonymous with formula weight in such cases.
Molar concentration, also called molarity, amount concentration or substance concentration is a
measure of the concentration of a solute in a solution, or of any chemical species, in terms of
amount of substance in a given volume.
Unit for molar concentration is defined as the number of moles per litre.

WEIGHT PERCENT

• This system of concentration is commonly employed to express approximate concentrations of

laboratory reagents.

• It specifies number of grams of solute per 100 g of solution:

P = wx100

w + wo

Where,

P is the percent by weight of solute,

w the number of grams of solute, and

wo the number of grams of solvent.

PARTS PER MILLION (PPM)

• PARTS PER MILLIONis convenient for expressing the concentrations of very dilute solutions.

67
• It specifies the number of parts of solute in 1 million parts of solution and can be expressed
mathematically as

ppm = w × 106

w + wo

Where,

w is the number of grams of solute

wo the number of grams of solvent.

STOICHIOMETRIC CALCULATIONS

Once the concentration of a solution determined, it can be employed as a titrant in the determination of
purity of an unknown sample. The calculations involved, called stoichiometric, are based on the mole
and mass relations between the elements and compounds as expressed by a chemical equation.

STANDARDIZATION OF SOLUTIONS

The process by which the concentration of a solution is determined is called standardization. The few
substances which are adequate in this regard are called primary standards. A solution is standardized by
a titration in which it reacts with a weighed portion of a primary standard. A widely used primary
standard for base solutions is the compound potassium hydrogen phthalate, abbreviated KHP, Sulfamic
acid, HSO3NH2, and potassium hydrogen iodate, KH(IO3)2 , are both strong acids and are excellent
primary standards. Sodium carbonate, Na2CO3 and tris(hydroxymethyl) aminomethane, known as
TRIS or THAM, are common primary standards for strong acids.

ALIQUOTS

Analyst weighs the primary standard, dissolves it in a volumetric flask, and withdraws a portion of the
solution using a pipette, the withdrawn portion is called an aliquot. An aliquot is a known portion of the
whole, usually some simple fraction. This process of dilution to a known volume and removing a
portion for titration is called taking an aliquot.

DILUTION

68
Take an aliquot of a standard solution and diluting it to a larger volume in a volumetric flask. The
number of moles of solute in the original solution must be the same as the number of moles in the final
solution.

EQUIVALENT WEIGHTS AND THE NORMALITY SYSTEM OF CONCENTRATION

The term equivalent in an attempt to simplify stoichiometric calculations in titrimetry. A titration

involves adding the titrant until an amount chemically equivalent to the analyte is reached and this point
is called the equivalence point (EPt). The equivalent is defined so that at the EPt the equivalents of
analyte and titrant are always equal. Examples:

HCl + NaOH ---------NaCl + H2O (1)

H2SO4 + 2NaOH ----Na2SO4 + 2H2O (2)

2HCl + Ca (OH)2 ------ CaSO4 + 2H2O (3)

H2SO4 +Ca(OH)2 ------ CaSO4 + 2H2O (4)

NORMALITY

The normality system of concentration is based on the volume of solution.

Normality = number of equivalents per liter of solution

N = eq/ V

Where N is the normality, eq the number of equivalents, and V the volume of solution in liters.

ACID-BASE TITRATION

Titrations involving acids and bases are widely employed in the analytical chemistry.

BRONSTED TREATMENT OF ACIDS AND BASES

The quantitative treatment of acid -baseequilibria became possible after 1887, when Arrhenius
presented his theory of electrolytic dissociation water solution, according to Arrhenius, acids dissociate
into hydrogen ions and anions, and bases dissociate into hydroxide ions and cations:

Acid: HX ---- H+ + X-

Base: BOH --------OH- + B+

69
The Debye-Huckel theory (1923) permitted a refined treatment that was even better.

The Bronsted Theory

In 1923, Bronsted presented a new view of acid-base behaviour. In Bronsted terms, an acid is any
substance that can donate a proton, and a base is a substance that can receive a proton. The hydroxide
ion, to be sure is such a proton acceptor and hence a Bronsted base. When an acid yields a proton, the
deficient species must have some proton affinity and hence it is a base. Thus, in the Bronsted treatment
we encounter conjugate acid-base pairs:

HB --------- H+ + B

The acid HB may be electrically neutral, anionic, or cationic (e.g., HCl, H2SO4, NH4 + ), and thus we
have not specified the charge on either HB or B.

LEVELING EFFECT

If HB is inherently a stronger acid than HS+, it will transfer its proton to the solvent

HB + S --------- HS+ + B

A series of acids, all of which are very much stronger than the solvated proton, will dissociate
completely; such solutions will be brought to a level of acidity governed by the acid strength of HS+ .
This is known as the leveling effect. According toBronsted terms, the dissociation of bases is treated in
a similar fashion, except that here the process is promoted by the acidity of the solvent.

TITRATION CURVES

To know about a reaction to determine whether or not it can be used for a titration, it is of interest to
construct a titration curve. For acid-base reactions a titration curve consists of a plot of pH or pOH vs.
milliliters of titrant. We shall examine two cases, titration of a strong acid with a strong base and
titration of a weak acid with a strong base.

Strong Acid-Strong Base Titration

Strong acids and bases are completely dissociated in aqueous solution. At the equivalence point the pH
is determined by the extent to which water dissociates at 25°C the pH of pure water is 7.00.

Weak Acid vsStrong Base Titration

70
Titration of a weak Acid with a strong base at the equivalence point, all of the weak acid is neutralized
and converted to its conjugate base (the number of moles of H + is equal to added number of moles of
OH - ). However, the pH at the equivalence point does not equal 7.

INDICATORS FOR ACID BASE TITRATION

Principle of indicator Behaviour

There are many weak organic acids and bases in which the undissociated and ionic forms show
different colors. Such molecules may be used to determine when sufficient titrant has been added and
are termed visual indicators. A simple example is p-nitrophenol, which is a weak acid.

The undissociated form is colorless, but anion, which has single and double bonds is yellow.
Phenolphthalein is adiprotic acid and is colorless. It dissociates first to a colorless form and then, on
losing the second proton, to an ion with a conjugated system; a red color result. Methyl orange
indicator, is a base and is yellow in the molecular form. Addition of a proton gives a cation which is
pink in color.

INDICATOR ERRORS

There are two chances of error in determination of the end point of a titration using visual indicators.
One occurs when the indicator does not change itscolour at the proper pH. This is a determinate error
and can be corrected by the running an indicator blank. The indicator blank is usually determined
experimentally. A second error occurs in case of weak acids (or bases) when we plot the curvewhere
slope of the titration curve is not great and hence color change at the end point is not sharp.

Selection of proper indicator

Name of indicator Indicator range

Methyl orange 3.1 to 4.4
Bromthymol 6.0 to 7.6
phenolphthalein 8.0 to 9.6

A strong acid is titrated, large change in pH at equivalence point is sufficient to span the ranges of all
three indicators. Hence anyone of these indicators would change colour within one or two drops of the
equivalence point, as would any other indicator' changing colour between pH 4 and 10. In titration of

71
weaker acids, choice of indicators is much more limited. Phenolphthalein changes colour at
approximately the equivalence point and is a suitable indicator.

MAGNITUDE OF THE EQUILIBRIUM CONSTANT

The concentrations of the substance titrated and the titrant influence the magnitude of ∆pH, and under
certain circumstances. All of the substance titrated be converted into product at or near the equivalence
point and conversion of the analyte into product at the equivalence point. It is also desirable that the pH
change by 1or 2 units on the addition of a few drops of titrant at the equivalence point if a visual
indicator is to be employed.

Effect of Concentration

The magnitude of ∆pH at the equivalence point also depends upon the concentrations (fig 4) of the
analyte and the titrant.

APPLICATIONS OF ACID-BASE TITRATIONS

Acid-base titrations are widely used for chemical analyses.

 Acid-Base Reagents
 In laboratory practice, to prepare and standardize one solution of an acid and one of a base.
 These two solutions can then be used to analyze unknown samples of acids and bases.
 Since acid solutions are more easily preserved than basic solutions.
 An acid is normally chosen as a permanent reference standard in preference to a base.
 Primary Standards
 In laboratory practice, it is customary to prepare solutions of an acid and a base of desired
concentration and then to standardize the solutions against a primary standard.
 Analyses Using Acid-Base Titrations
 A wide variety of acidic and basic substances, both inorganic and organic, can be determined by
an acid-base titration.
 SOLVENT SYSTEMS

Several classifications of solvents have been proposed. Some, such as methanol and ethanol, have acid-
base properties comparable to water and, along with water, are called neutral solvents. Others, called
acid solvents, such as acetic acid, formic acid, and sulfuric acid, are much stronger acids and weaker
bases than water. Basic solvents such as liquid ammonia and ethylenediamine have greater basicity and

72
weaker acidity than water. Aprotic, or inert, solvents are neither appreciably acidic nor basic and hence
show little or no tendency to undergo autoprotolysis reactions. Pyridine, for example, can accept a
proton from an acid such as water:

Pyridine has no tendency to furnish a proton. A fourth class of solvents would be with acidic but no
basic properties.

The titrant is the solution used in a titration to determine the concentration of an unknown solution. The
titrant is a solution of known concentration that is by a burette delivered into a known quantity of the
solution of unknown concentration. Perchloric acid is' by far the most widely used acid for the titration
of weak bases.

END-POINT DETECTION

A number of visual indicators are available such as cresol red methyl red, azo violet and crystal violet.
The rationale of indicator selection does not have a good theoretical base, and the choice is often best
made on the basis of experience, trial and error. Other instrumental end points such as conductometric
and photometric have been used successfully.

Chapter 9 Acids, bases and their salts and complexes

Introduction

The properties and reactions of aqueous solutions of acids and basesare fundamental to all branches of
chemistry and related sciences. The free hydrogen ion concentration dominates many chemical
reactions by determining the extent to which the reaction proceeds, the rate at which it goes, or the
detailed mechanism by which it takes place.

Acid-base concepts

The titration of a base with an acid is one of the simplest of all analytical determinations, and is one of
the first experiments to be studied by the student. Here, we shall study a summary of the definitions and
elementary principles, and then consider the complex equilibria that prevail in solutions of polyprotic
systems, and the use of solvents other than water.

73
The common definitions for acid-base systems are in the below table. For our purposes, the Bronsted
concept is the most useful. In this model, when an acid donates its proton, the remaining fragment
becomes the conjugate base of the original acid.

Originator Acid Base Neutralization

Arrhenius Provides H+ in water Provides OH- in water H+ + OH ---> H2O
Brownsted – Lowery H+ donor electron pair H+ acceptor electron Proton transfer
lewis acceptor pair donar formation of a
coordinate- covalent
bond

If the conjugate base accepts a proton, it reverts to the parent acid. Likewise, every base has a conjugate
acid.

Role of the solvent

Free protons do not exist under normal conditions; therefore, an acid cannot donate its proton unless a
base is present to accept it. Many solvents, including water, possess basic properties, that is, they can
accept protons. Thus, when pure HCl is dissolved in water, a proton transfer reaction takes place:

HCl + H2O ------- H3O + + Cl-

acid 1 base 2 acid 2 base 1

Acetic acid, CH3COOH, is abbreviated as HOAc. In the above reaction, the OHand the OAcions
compete for the proton. The OHion is a much stronger base than OAcion, therefore the equilibrium lies
far to the right. A comparison of the two reactions just given shows that water can act as either an acid
or a base depending on the acidic or basic nature of the other partner in the reaction-water is an
amphoteric substance, and it is also an amphiprotic solvent.

Amphiprotic solvents undergo autoprotolysis, or self-ionization, a proton transfer reaction where one
solvent molecule (acting as an acid) donates a proton to another solvent molecule (acting as a base).
Water is the most familiar example, but there are many others:

H2O + H2O ------H3O + + OH

74
acid1 base2 acid2 base1

Classification of Solvents:

Solvents are classified with respect to acid-base properties as either aprotic (inert) or amphiprotic.
Aprotic solvents are neither acidic nor basic (e.g., benzene and carbon tetrachloride). Amphiprotic
solvents act as both proton acceptors and proton donors. There are gradations among the amphiprotic
solvents, from predominantly acidic to predominantly basic. Glacial acetic acid is very acidic, whereas
liquid ammonia is very basic. Water and ethanol are neither strongly acidic nor strongly basic.
Protogenic amphiprotic solvents (e.g., sulfuric and formic acids) exhibit very strong acidic properties
and very weak basic properties. Autoprotolysis constants for protogenic solvents are usually larger than
that of water (e.g., Ks = 10-6 for formic acid, whereas Ks = 10-14 for water). Intermediate amphiprotic
solvents possess weakly acidic protons and can also act as very weak bases. Autoprotolysis constants
for nonaqueous intermediate amphiprotic solvents tend to be smaller than that of water (e.g., Ks = 10-
19 for ethanol). Protophilic amphiprotic solvents (e.g., ethylenediamine) exhibit very weak acidic
properties and relatively strong basic properties; their autoprotolysis constants are usually less than that
of water.

Dielectric Constant Effects:

The strength of an acid, HB, in a given solvent, SH, is defined in terms of the extent to which the
reaction HB + SH SH2 + + B- proceeds. This reaction is a combination of two steps, ionization and
dissociation:

HB + SH --------- SH2 +B - ----------SH2 + + B

In aqueous solutions both of these steps occur rapidly and aqueous acid-base reactions are thus very fast
reactions. In any solvent, the extent of the ionization step depends on the relative strength of the
conjugate acid-conjugate base pairs. The extent of the dissociation step depends on the charge type of
the members of the ion-pair and the polarity of the solvent. The dielectric constant is a measure of this
polarity-the higher the dielectric constant, the more polar the solvent. The extent of dissociation of ion-
pair aggregates increases with the dielectric constant of the solvent. Clearly then, solvents with high
dielectric constants are necessary for complete dissociation. The dielectric constant of a vacuum is
arbitrarily defined as zero. The very polar water molecule has a dielectric constant of 78.5 whereas the
value for the slightly polar acetic acid molecule is 6.2

75
Relative strengths of acids and bases

The strengths of different acids can be measured by comparing their ability to donate a proton to some
common base. Likewise, the strengths of different bases can be measured by comparing their ability to
accept a proton from some common acid. Water is selected as both the common acid and the common
base for these comparisons. The quantitative measure of these strengths is the equilibrium constant for
the appropriate proton transfer reaction. For acids we use Ka, the acid dissociation or "acidity" constant.
For bases we use Kb, the base dissociation or "basicity" constant:

HCl + H2O → H3O + + Cl

Water is also the solvent and is present in large excess. Therefore, its concentration remains constant.
By convention, the activity of a pure solvent is taken as unity, and its concentration does not appear in
the mass action expression.

The relative values of the Ka's and Kb's indicate that hydrochloric acid is a stronger acid than acetic
acid, acetic acid is stronger than phenol (carbolic acid), and ammonia is a stronger base than aniline.

Equilibria in monoprotic systems

Calculating the hydrogen ion concentration (or pH) in a number of acid-base systems in water give us a
means of estimating the pH in various solutions including buffers and titration mixtures.

The following conventions have been used

1. The hydronium ion, H. (H2O) n + or H30 + , is used interchangeably with the hydrogen ion, H+ .

2. Brackets around a chemical formula indicate the molar concentration of that species. The activity
coefficient is ignored (i.e., assumed to be unity), because in practice numerical values of activity
coefficients are rarely available.

3. Equilibrium constants are assumed to be constant even though the distinction between activity and
concentration is not made. These approximations are adequate for present purposes. The temperature is
assumed to be 25°C.

4. pH = -log [H + ]; pOH = -log [OH- ]; pKw = -log Kw therefore pH + pOH = pKw = 14.060 at 25°C.

pH Calculations in Simple Systems: In calculating the [H+ ] or pH of solutions of monoprotic acids,

bases, and their salts, or mixtures thereof, the main difficulty arises in deciding which of the various

76
possible species are the principal constituents of the solution. Once this has been determined, the
appropriate equation is used:

1. Solution of a strong acid,

HX [H+ ] = CHX (1)

2. Solution of a weak acid,

HB [H+ ] = √ ka (CHB – [H+ ] ≈ √ Ka CHB (2)

3. Solution of the salt of a weak acid,

NaB [OH- ] = √ Kw / Ka (CB - - [OH] ) ≈ √ Kw /Ka CB - (3)

Monoprotic Weak Acid Dissociation Constants at 25oC

Acid name formula Ka pKa

Acetic CH3COOH 1.75 X 10-5 4.76
Benzoic C6H3COOH 6.3 X 10-3 4.20
Boric HBO2 5.8 X 10-16 9.24
Chloroacetic ClCH2COOH 1.38 X 10-3 2.86
Cyanic HOCN 2.0 X 10-4 3.70
Formic HCOOH 1.77 X 10-4 3.75
Glycolic HOCH2COOH 1.32 X 10-4 3.88
Hydrocyanide HCN 4.8 X 10-16 9.32
Hydrofluroric HF 6.75 X 10-4 3.17
Hypochlorous HOCl 2.95 X 10-8 7.53
Lactic CH3CHOHCOOH 1.38 X 10-4 3.86
Nitrous HNO2 5.1 X 10-4 3.29
Phenol C6H5OH 1.05 X 10-10 9.98
Propionic CH3CH2COOH 1.34 X 10-3 4.87
Ammonium ion NH+4 5.62 X 10-10 9.25
Pyridium ion C3H3NH+ 6.2 X 10-6 5.21

77
4. Solution containing both a weak acid and its salt,
HB+ NaB, a buffer [H+ ] = Ka CHB / CB – (4)
OR pH = pKa + logCB - / CHB (5)
5. Solution of a weak base, BOH
[OH- ] = √ kb (CBOH – [OH- ] ≈ √ KB CBOH (6)
6. Solution of the salt of a weak base,
BX [H+ ] = √ Kw / Kb (CBx - - [H+ ] ) ≈ √ Kw /Kb CBx (7)
7. Solution containing both a weak base and its salt,]
BOH + BX, a buffer [OH- ] = Kb CBOH/ CBX (8)
Equations 5 and 9 are known as the Henderson-Hasselbalch equations. All of the above equations
neglect the contribution to [H+ ] and [OH- ] provided by the autoprotolysis of water. These
contributions, which can never exceed 10-7 M, are too small to take into account except in highly
dilute solutions. Following examples will illustrate the application of the above equations.
Buffer Solutions:
Many chemical reactions generate free hydrogen or hydroxyl ions. If these ions remained in the
system, there would be significant changes in the pH. Buffer solutions contain constituents that
react with both strong acids and strong bases in such a way that the free hydrogen ion concentration
of the system remains relatively constant. Buffers typically consist of mixtures of weak acids and
their conjugate bases, or of weak bases and their conjugate acids.
Buffer Capacity:
Buffer solutions resist changes in pH upon the addition of strong acids or strong bases. The buffer
capacity of a system is defined as the moles of strong acid or strong base required to change the pH
of 1 liter of the buffer solution by 1 unit. The larger the buffer capacity, the better the buffer, that is,
the more acid or base it can consume without significant changes in pH. Clearly, more concentrated
buffer solutions have higher capacity than more dilute solutions. For example, even though a
solution that is 0.0100 F in both acetic acid and acetate ion has the same pH as a solution which is
0.100 F in both of these constituents, the more concentrated solution can consume ten times as
much strong acid or strong base for the same change in pH.
Acid-base titration in aqueous solution
One of the principal reasons for mastering pH calculations is to be able to use them in predicting
and analyzing titration curves for acid-base reactions. A titration curve shows how the pH of a
solution changes upon addition of acid or base; it is a plot of pH versus volume of titrant added. The
region near the equivalence point is of special interest. The inflection point, which occurs at the
78
steepest portion of the curve, coincides with the equivalence point. The steeper the curve, the
sharper the end point and the more accurate the titration will be. Titration curves be obtained
experimentally by measuring the pH with a pH meter after adding successive increments of titrant.
The routine steps to follow in plotting any titration curve are:
1. Write the chemical reaction for the titration.
2. Select the initial concentration, Co , and volume, Vo , of the sample to be titrated.
3. Select several volumes of titrant added, V, and calculate the fraction, f, of the sample which
is titrated at each point, f = V / VeP, where VeP is the volume of titrant required to reach the
equivalence point. It is convenient to select values of V such that f = 0, 0.1, 0.5, 0.9, 1.0, 1.1,
and 1.5.
4. Determine the concentration of the principal species present at each f value selected and
compute the pH.
5. Tabulate and plot the points selected.

Acid-Base Indicators:

Solutions of the common acids and bases are colorless, thus colored indicators are useful to detect
the end points of their titration. Acid-base indicators are intensely colored weak acids (or weak
bases) whose conjugate form has a different color. A small amount of the indicator is added to the
sample and is titrated along with it. An indicator is selected that is just half-titrated at the pH of the
equivalence point of the main titration. The indicator will exist half in its acid form and half in its
basic form and exhibit an intermediate color. This results if the pKa of the indicator is close to the
pH at the equivalence point, pHep

pHep = pKa + log CB / CA

Table :Typical Acid – Base Indicators

Indicator Acid form p H transistion Base form

Paramethyl red Red 1.0 to 3.0 Yellow
2,6 – Dinitrophenol Colorless 2.0 to 4.0 Yellow
Bromophenol blue Yellow 3.0 to 4.6 Blue
Methyl orange Red 3.1 to 4.4 Yellow
Bromocresol green Yellow 3.8 5.4 Blue
79
Bromomothymol blue Yellow 4.2 6.2 Yellow
Phenolphthalein Colorless 5.2 6.8 Purple
Thymolphthalein Colorless 6.0 7.6 Blue
Alizarin yellow R Yellow 8.0 9.6 Red
2,4,6 – Trinitrotoluene colorless 9.3 10.6 blue

Acid-base titrations in non-aqueous solvents

Most of the non-aqueous titrations involve neutralizations of organic bases or acids and the direct
determinations of acidic and basic functional groups. In addition to their importance in organic
analysis, non-aqueous acid-base titrations are used extensively in pharmaceutical analysis, for
example, to determine constituents present in antihistamines, antibiotics, and sulfonamides. Direct
titrations are possible in non-aqueous acidbase reactions since they proceed to completion within a
few microseconds.

There are numerous advantages offered by acid-base titrations in non-aqueous media, the most
important of which is that a much larger number of acids and bases can be titrated in nonaqueous
solvents than in aqueous solution. For example, a weak acid with a pKa of 9 or greater cannot be
determined accurately in water because of the competition of the solvent for the strong base titrant
(leveling effect). But the weak acid can be titrated in ethylenediamine, which is considerably more
basic than water. The availability of a large variety of non-aqueous solvents allows the choice of a
solvent that will assist but not interfere with a specific acid-base titration. A properly formulated
non-aqueous acid-base titration procedure provides results that are very accurate and often more
precise than those obtained from a corresponding titration in aqueous media. We will consider
briefly some of the practical considerations that apply to non-aqueous acid-base titrations.

Titrants: Since perchloric acid is the strongest mineral acid, acidic titrants normally consist of
solutions of perchloric acid in either anhydrous acetic acid or dioxane. Acetic acid solutions that are
0.1 to 1.0 Fin perchloric acid commonly are used. Perchloric acid is essentially 100% ionized in
acetic acid; it exists as the ion pair, H2OAc+ClO4 - as well as the solvated ion, H2OAc+ .

Basic titrants include quaternary tetraalkyl ammonium hydroxides, which are stronger bases than
the alkali metal hydroxides. Although the product salts of alkali metal ions are quite insoluble in

80
non-aqueous media, quaternary ammonium salts are very soluble. Tetrabutylammonium hydroxide
in isopropanol is the most commonly used base titrant. To insure that the titrant solvent does not
interfere with the principal reaction, the concentration of the titrant solution is usually high relative
to the concentration of the substance titrated. Only a small volume of titrant is added compared to
the volume of the titration mixture.

Choice of Solvent for Acid-Base Titration: The following considerations are pertinent to the
choice of a solvent for a specific non-aqueous acid-base titration.

1. The solvent should permit a large change in the solvated proton concentration near the
equivalence point. Other things being equal, the smaller the autoprotolysis constant, the better the
end point.

2. The substance to be titrated must be soluble, either in the solvent or in an excess of the titrant
which then may be back-titrated.

3. The product of the titration must be soluble, or if it is a precipitate, it must be compact and
crystalline and not gelatinous. Gelatinous precipitates tend to interfere with accurate end-point
determinations.

4. The solvent should not introduce interfering side reactions with either the substance to be titrated
or the titrant.

5. Preferably, the solvent should be inexpensive and easily purified. Many of these solvents are
toxic and require special precautions in handling.

Inert solvents may be added to either protogenic or protophilic solvents to modify differentiating
titrations. The presence of the inert solvent decreases the amphiprotic character of both acidic and
basic solvents. For example, butylamine and pyridine are both titrated as strong bases in glacial
acetic acid, but in a solvent which consists of 10% acetic acid and 90% chloroform, two distinct
equivalence points are noted corresponding to the neutralizations of butylamine and then pyridine.

Normally, the titration of a weak acid in an inert solvent involves the addition of a small volume of
concentrated base dissolved in an amphiprotic solvent such as an alcohol. Thus, after the first
addition of titrant, the system contains a small amount of amphiprotic solvent (the alcohol) in an
inert solvent. The "effective" autoprotolysis constant for the system is that of the amphiprotic
solvent mixed with the inert solvent. When the amount of amphiprotic solvent is small, the

81
"effective" autoprotolysis constant is small and the "pH" break is large near the equivalence point.
This is the reason why highly concentrated solutions of bases are used as titrants. Isopropyl alcohol
is commonly used as the solvent for tetra-alkylammonium hydroxides because it is the least acidic
of the amphiprotic solvents.

In summary, for titrations of weak acids and weak bases, inert solvents provide the largest break in
"pH" near the equivalence point. Inert solvents are also preferable in successive titrations of acids
and bases in mixtures. When solubility problems are encountered with inert solvents, protogenic
solvents are used for weak base titrations and protophilic solvents are used in weak acid titrations.

End Point Determination in Non-aqueous Titrations: Unfortunately, in non-aqueous media we

do not have accurate values for the dissociation constants of chemical indicators, nor do we have
quantitative relationships which relate the e.m.f. of electrochemical cells to constituent
concentrations. Let us briefly examine some of the indicator systems for non-aqueous titrations.

Potentiometric End Point: In a typical non-aqueous potentiometric acid-base titration, we might

use this cell: Glass Membrane Ag ‫׀‬AgCl, HCl (0.10 N) :Non-aqueous‫ ׀׀‬Saturated Calomel Solution
Electrode.

As in aqueous solution, in most non-aqueous solvents the glass electrode appears to respond to the
difference between the activity of the solvated proton in the solution inside the glass membrane and
the activity of solvated proton in the solution on the outside. However, a quantitative interpretation
of this potential difference is not obvious. In the first place, ion activities in one solvent cannot be
simply related to ion activities in another solvent. In addition, when we measure the e.m.f. of the
cell just shown, we include a sizable, unknown, liquid junction potential between the aqueous
calomel electrode and the non-aqueous solution. This liquid junction potential may vary
considerably during the course of a non-aqueous titration, depending on the nature of the solvent
and the titrants. Liquid junction potentials must remain reasonably constant during the course of the
titration for high precision. In many cases the e.m.f. of the above cell does vary approximately
linearly with the pSH2 of the non-aqueous solution. It should be pointed out, however, that the
e.m.f. variation in no way corresponds to the theoretical variation derived for aqueous solutions
from the Nernst equation. In other words, a change in 1.00 unit of pSH2 does not correspond to a
change in e.m.f. of 59.16 mV at 25°C in non-aqueous solvents. As a result, the pH scale of a pH
meter is meaningless in nonaqueous titrations and the millivolt scale(s) should be used. So, for
titrations in ethanol or glacial acetic acid, the glass-calomel electrode system appears to respond to
82
changes in -log [EtOH2 + ] and -log [H2OAc+ ], respectively (or the "pH" of these solutions).
When an acid is titrated in an inert solvent (e.g., a ketone) with t-butyl ammonium hydroxide in
isopropanol, the glass electrode responds to changes in the isopropanol solvated proton
concentration. In highly basic solvents, such as butylamine and ethylenediamine, the glass electrode
does not function as an indicator electrode and an antimony electrode is used instead.

Chemical Indicators: Indicators for non-aqueous acid-base titrations behave similarly to those that
are employed in aqueous titrations in either giving up or accepting protons and changing color in the
process. An indicator, therefore, may dissociate in solvent

SH HIn + SH ------- SH2 + + In color A color B

Clearly the predominant form of the indicator depends on the SH2 + concentration. So far, this story
is no different from that for an indicator in aqueous solution. However, in non-aqueous media many
solvents are available and the behavior of a particular indicator depends on the acidity, basicity, or
inertness of the solvent as well as its dielectric constant.

Some indicators have simple color changes; others successively pass through a wide range of color
shades. For example, as crystal violet is increasingly protonated in glacial acetic acid, it changes
from violet to blue-green to green to yellow. Methyl violet passes through similar changes. To
determine the color change that best indicates the end point, a potentiometric titration should be run
with the indicator present. The color change that occurs nearest the potentiometric equivalence point
should then be used as the end point in future titrations of the same kind. A list of indicators for
titrations of weak acids and weak bases is given in Table

Indicator Constituent titrated Solvent

Crystal violet or methyl violet Weak bases Acetic acid; acetonitrile
Methyl red Weak bases Dioxane; glycol - isopropanol
Dibenzalacetone Weak bases Nitromethane
Azo-violet Weak acids Dimethylformaide
Or
Thymol blue
Or
o- Nitroaniline

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 CHAPTER 10

GRAVIMETRIC METHODS OF ANALYSIS

Introduction

Gravimetry is process of isolation of a particular compound of the element in form of precipitate

and then taking its weight as possible as pure. The compound is isolated from a weighed portion of
the substance being analyzed. The element weight can be easily calculated from the formula of the
analyte and the relative atomic weights of the constituent compounds are known.

Gravimetric procedure advantages:

(a) Gravimetry is accurate and precise if properly calibrated electronic balances are employed;

(b) Errorsare easily checked as tests are carried out on filtrates for completion of precipitation
reaction and examination of obtained precipitates can be done forpresence of various impurities;

(c) Gravimetry is an absolute method, i.e. itinvolves direct measurement and there is no need of any
form of calibration;

(d) Gravimetry experimentscan be done with cheap apparatus.

Method of precipitation

The analyte which is being quantified is precipitated in a form which is very slightly soluble and
thus there is no considerable loss when the precipitate is filtered from the solution. The factors
which are responsible for a successful determinationare:

1. The formed precipitate should be insoluble that there is no considerable loss as collected by
filtration.
2. The obtained precipitate must possess a physical nature. 3. The precipitate should easily get
converted into a pure substance. Previously it was thought that the precipitate which was
obtained was chemically pure and it didn’t contain any impurities but this is not always right.
Purity ofprecipitate is dependent upon the various components in the solution and the conditions
of precipitation.

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Colloids

Colloidal particles range between 0.1 µmand 1 nm. These particles are not filterable. When beam of
light is passed through colloidal solution and then the solution is observed at right angles to the incident
light, a light of scatter is seen. This effect is known as Tyndall effect. Colloids are of two types:
lyophobic and lyophilic colloids. Dispersing a gel or flocculated solid to form sol is the process
Peptization. Stability oflyophobic colloids depends on the electrical charge on the particles. When
arsenic (III) sulphide solis formed by precipitation process with hydrogen sulphide in solution
containing an acid, the sulphide ions getadsorbed first, and then some hydrogen ions get adsorbed on the
primary layer of sulphide ions. The hydrogen ionswhich are secondarily adsorbed are known as 'counter-
ions'. Therefore, a double layer is formed. Surface of colloidal particles of arsenic(III)
sulphideisnegativelycharged, with positively charged counter-ions. These counter-ionsimpart
positivecharge to the liquid which surroundsthem. When current is passed through the solution, negative
charged particles shift towards anode. If electrical double layer gets destroyed, the sol becomesunstable,
and flocculation of the particles is seen, and thus surface area is reduced. Therefore, if solution of
bariumchloride is added, barium ions get adsorbed by theparticles; distribution of charge on surface gets
delivered and particles flocculation is observed. Minimum quantity of electrolyte which is required to
flocculatethe colloidal particles is known ascoagulation or flocculation value. Insuchsituations,
theprecipitate gets contaminated by adsorption of foreign materials on its surface. If the precipitate is
washed with water to remove the adsorbed substances, then a new problem arises. The concentration of
the electrolyte presents in the supernatant falls below coagulation value, and precipitate passes into
colloidalsolution again. This is known as peptisation, and this phenomenon is of great importance in
gravimetric determinations.

Supersaturation and formation of precipitate

Supersaturated solution is a solution containing a higher concentration of solute than correlate to

equilibrium solubility at a given heat. Supersaturation is an unstablestate and equilibrium stability can be
achieved by adding crystal of solute or of any other compound, ormechanically by means of shaking or
stirring. Von Weimarn stated that supersaturation plays a crucial role in determination of the size of
particle precipitate. He found out that the initial velocity of precipitation is proportional to (Q - S)/S,
where Q is total concentrationof substance whose precipitation is to be done, and S is equilibrium
solubility; (Q - S) indicates the super saturation at the moment precipitation starts. To obtain a

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precipitate with crystalline nature, (Q - S)/S should be small. Points to be considered before carrying out
process of precipitation are:

1. Hot solutions are generally employed to carry out precipitation, because solubility usually increases
with increase in temperature.

2. Precipitation is usually done in dilute solutions and precipitating reagent is slowly added with proper
mixing. Addition of reagent slowly if necessary

3. Process which is generally used to avoid supersaturation is doing precipitation from a homogeneous
solution. In this procedure the precipitating agent is generated within the solution.

Co-precipitation When a precipitate is formed and is separated by filtration, it contains some impurities:
itmay retaindifferentquantities of contaminants depending upon nature of precipitate and experimental
circumtances of precipitation. Precipitate infection by materials soluble in mother liquor istermed co-
precipitation. There are two types of co-precipitation. First one type is the contamination due to
adsorption at surfaceof particles of the precipitate which are exposed to the solution, and the second type
is concerned with the occlusion ofimpurities during growth of crystal from the nuclei. Contamination of
gelatinous precipitates is an example of surface adsorption. The another type of co-precipitation,
occlusion, occurs duringbuilding up of precipitate from nuclei. In addition to co-precipitation,
postprecipitationis also responsible for introduction of errors. Post-precipitation occurs on surface of
first precipitate afterit has been formed. When calcium is precipitated in the form of oxalate in presence
of magnesium, then magnesium oxalate is also formed which gets deposited on calcium oxalate
precipitate and contaminates the precipitate. Longer the precipitate is kept in this solution greater will be
the contamination.

Post-precipitation is different from co-precipitation in many ways:

 Contamination in case of post precipitation increases with time if the precipitate is allowed to
remain incontact with mother liquor, but in co-precipitation contamination decreases with time.
 Contamination in case of post precipitation increases if the solution isstirredvery fast.
Thisdoesn’t happen in case of co-precipitation.
 Magnitude of contamination is found to be much greaterin post precipitation. The process of
digestion influences the quality of the precipitate to a great extent.Digestion is generally done by
leaving the precipitate undisturbed for some hours in the presence of mother liquor, the process
is generally carried out at room temperature. The aim of this process is togeta precipitate in a

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form whose filtration can be done very easily so that the loss during filtering process is
minimized. Two changes are seen when the precipitate in contact with mother liquor. The small
particles having much greater solubility than the large particles Analytical Chemistry /
Instrumentation Fundamentals of Analytical Chemistry Gravimetric methods of analysis have a
tendency to pass into the solution, and these small particles with time re-deposit upon
largeparticles. Crystals are formed very rapidly and possess a very irregular shape and a large
surface; after digestion or ageing of the precipitate these crystals become denser with a decreased
surface area. Digestiongenerally reduces co-precipitation and increases size of thecrystals, so that
their filtration is easy.
Requirements for precipitation
a. Precipitation is usually done with dilute solutions. Solubility of precipitate, time needed to
filter the precipitate filtration and manydifferent stepswhich are carried out after obtaining the
precipitate as filtrate should be paid due attention. All this will be of immense help in reducing
the errors. b. Mixing of the reagents should be done very slowly continuous stirring. This helps
in keeping the degree of supersaturationlow and will also help in formation of largecrystals. The
precipitating reagent is always added in a slight excess to ensure complete precipitation.
Inmanycases order of mixing the reagents plays an important role.
c. Rate of precipitation increases in hot solutions. When temperature is high:
(i)An increaseinsolubility is seen whilesupersaturation decreases,
(ii) coagulation is enhanced decreasing formation of sol and
(iii) rate of crystal formation also increases, which leads to large and regular formed crystals.
d. Digestion of precipitates of crystalline nature should be done for as long as possible, the
precipitate is usually kept overnight to obtain a good precipitate with large particle size. Keeping
the precipitate for several hours for digestion is avoided in cases where postprecipitation may
occur.
e. Washing of the precipitate is done with asuitableelectrolyte solution. Washing with pure water
is avoided as it may causepeptisation.
f. If considerable contamination is still observed in the obtained precipitate, then, the precipitate
should be dissolvedin anappropriate solvent and then re-precipitation is done. The quantity of
impuritiesin the precipitate after re-precipitation is considerablyreduced and thus more reliable
results can be obtained.
Filtration of the obtained precipitate:

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Filtration of the precipitate is done precipitation and digestion are complete. Filtration is done
generally by using filter paper or filtering crucible. The most widely used medium for filtration is
a cellulose-based filter paper. As a filter paper is hygroscopic it is very difficult to dryit to a
constant weight. Therefore, in a quantitative determinationitis removed before taking weight of
the precipitate. This is done by ignition of the filter paper with great care. Afterigniting the filter
papersome amount of inorganic ash is left behind whichis responsible for erroneous resultsin the
final weight of the precipitate. Therefore, to reduce errors due to the non-combustible ash a low-
ash filter paper is employed. This type of filter paper is first treated by washing with an
acidmixture for removal of inorganic substances. Filtration is done by folding filter paper in
shape of a cone. This folded filter paper is then placed in a funnel. Then the filter paper is
moistened with water and the filter paper is pressed to the wall of the funnel. All of the
precipitate is not transferred at a single time. Small portions of the precipitate are transferred in
several steps. The first step is to remove a large amount of supernatant through filter paper
without the transfer of the precipitate. This prevents clogging of the filter paper. Another
medium of filtration is a filtering crucible. The most widely used crucible is a fritted glass
crucible which contains a porous disk filter made of glass. Gooch crucible is also a type of
filtering crucible having perforations at the bottom. The precipitate in this crucible is retained by
the glass fiber mat. The supernatant is decanted with the help of a vacuum pump.
Washing
Filtering aids in removal of most of the supernatant. But some amount of the supernatant, is
retained which should be removed to prevent introduction oferror in the result obtained.
Precipitate is rinsed carefully, without appreciable loss of the precipitate, to eliminate the
residual substances. To minimize solubility losses during rinsing of the precipitate, coldsolvents
or solutions which containorganic solvents are used as rinsing agents. pH of the
precipitateswhich contain acidic or basic ions should be properly adjusted as an improper pH
may lead to solubility losses. In some cases, where coagulation plays a crucial role in
determination of the particle size, addition of an electrolyte to rinse solution is done
toavoidreturning backof the precipitate in a state where the filtering device becomes unable to
retain the precipitate. This process is known aspeptization. The electrolyte employed for this
purpose should be volatile in nature and is easily removed when the precipitate is dried.
Drying
After separation of the obtained precipitate from its supernatant, drying of the precipitate is done
to eliminate traces of rinsingagentand contaminants which are volatile in nature. Temperature
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and procedure adopted fordrying depends onfiltration methodas well as the desired chemical
form of the precipitate. Generally, a temperatureof 110 °C is employedto removemoisture and
other volatilecontaminants. A drying oven serves this purpose efficiently. When a high
temperature isrequired then a muffle furnace or a bunsen burner is used. To make sure that the
precipitate has been completely dried the precipitate is dried many times and weighed till the
analyst obtains a constant weight. Fritted glass crucibles are sensitive to high temperatures
andthey can’t be employed for drying purposes which require temperature above 200 °C.
Composition of precipitate obtained after drying
Quantitative determination through gravimetric analysis requires that precipitate obtained
shouldhave a definite composition. Precipitates which contain volatile ions or appreciableamount
of moisture are generally dried by supplying a temperature enough for complete removal of the
volatile substances. For example, in determination of magnesium through gravimetry, a
precipitateof MgNH4PO4 × 6H2O is formed. This precipitate can’t be dried at lowtemperatures
without loss ofsubstantial amounts of ammonia and water. Instead, drying of the precipitate is
done at temperatures above 1000°C. The precipitate at such a high temperature gets
decomposedto magnesium pyrophosphate.
Applications of gravimetry Some of the applications of gravimetry are given below:
Inorganic analysis The most popular precipitating reagents for inorganic cations aresulfide,
oxalate, chromate, sulfate, halides and phosphate. Same reactions can be used for determination
of various inorganic anions by simple reversal of the analyte and the precipitating reagent.
Organic analysis Gravimetry is very useful in determination of different organic functional
groups. Quantitative calculations Stoichiometry is employed to determine relationship
betweenanalyte and the precipitate obtained in a reaction.
Qualitative applications Gravimetry methods have been widely used for qualitative
determination of inorganic andorganic analytes.
Evaluation of gravimetry Scale of operation: Scale of operation for precipitation
gravimetrydepends on sensitivity of analytical balance and sample availability. For an
accuracyof ±0.1% employing a balance having a sensitivity of ±0.1 mg, the weight of the
precipitatemust be at least 0.1 g. Due to this limitation, samples containing analyte in trace
amounts can’t be determined with high accuracy.
Accuracy: Relative errors of 0.1–0.2% are usuallyseen. When there is a difficulty in obtaining a
pure precipitatefree from contaminants, then standardization is done to determine the relationship
between mass of the precipitate andmass of the analyte.
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Precision: Precision obtained for a particular analysis is dependent on thequantity of sample and
precipitate.
Selectivity: Precipitating reagents generally are not selective for a single analyte. For example,
silver is not a selective precipitating reagentfor chloride as it can form precipitate with bromide
and iodide also. Time, cost and equipment Gravimetric methods are time-consumingand are
rarely used when large number of samples are to be analyzed. Few equipments are needed to
carry out the analysis. The technique is not expensive and does not require skilled professionals.

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