Fundamental Analytical Chemistry Module - Latest Edition Dec, 2019

Basic Analytical Chemistry as
a Tool for Instrumentation

Training
JIJE Laboglass Pvt. Ltd. Company

ADDIS ABABA, ETHIOPIA
DECEMBER, 2019
Table of Contents
CONTENTS PAGE
LIST OF TABLES ................................................................................................................................................II

1. INTRODUCTION TO ANALYTICAL CHEMISTRY ............................................................................1
1.1. THE NATURE OF ANALYTICAL CHEMISTRY........................................................................................1
1.2. THE ROLE OF ANALYTICAL CHEMISTRY ............................................................................................4
1.3. CLASSIFICATION OF CHEMICAL ANALYSIS .........................................................................................5
1.4. METHODS OF ANALYSIS .......................................................................................................................9
1.4.1. Gravimetric Methods of Analysis .....................................................................................................9
TYPES OF GRAVIMETRIC METHODS ..........................................................................................................9
1.4.2. Titrimetric Methods of Analysis .......................................................................................................9
TYPES OF TITRIMETRIC METHODS .......................................................................................................... 10
1.4.3. Spectroscopic Methods of Analysis ................................................................................................ 10
SPECTROSCOPY BASED ON ABSORPTION ............................................................................................... 11
SPECTROSCOPY BASED ON EMISSION ..................................................................................................... 11
SPECTROSCOPY BASED ON SCATTERING ............................................................................................... 11
1.4.4. Electrochemical Methods of Analysis ............................................................................................. 12
1.4.5. Chromatographic and Electrophoretic Methods of Analysis ......................................................... 12
1.5. BASIC TERMINOLOGIES IN ANALYTICAL CHEMISTRY ..................................................................... 13
1.6. Error in Chemical Analysis ............................................................................................................ 17
1.6.1. Types of Error ................................................................................................................................ 17
1.6.1.1. Random (Indeterminate) Errors ........................................................................................................... 17
1.6.1.2. Systematic (Determinate) Errors .......................................................................................................... 18
1.6.1.3. Gross Errors ......................................................................................................................................... 19
2. CONCENTRATION AND METHODS OF EXPRESSING CONCENTRATION .............................. 19
2.1. MOLARITY (M) ................................................................................................................................... 19
2.2. NORMALITY (N).................................................................................................................................. 20
2.3. MOLALITY (M) .................................................................................................................................... 20
2.4. MOLE-FRACTION (X) .......................................................................................................................... 20
2.5. PERCENTAGE CONCENTRATION ........................................................................................................ 20
2.6. WEIGHT/ WEIGHT (W/W %) .............................................................................................................. 21
2.7. VOLUME/ VOLUME (V/V %) ............................................................................................................... 21
2.8. WEIGHT/ VOLUME (W/V %)............................................................................................................... 21
2.9. PARTS PER MILLION (PPM) ................................................................................................................ 21
2.10. PARTS PER BILLION (PPB) .................................................................................................................. 22
2.11. FORMAL SOLUTION ............................................................................................................................ 22
2.12. EQUIVALENT WEIGHT ....................................................................................................................... 24
3. VARIABILITY IN EQUIVALENT WEIGHTS ...................................................................................... 24
3.1. EQUIVALENT WEIGHT AND VALENCY ............................................................................................... 25
3.2. EQUIVALENT WEIGHT OF ACID, BASE AND SALT ............................................................................. 25
3.3. GRAM-EQUIVALENT WEIGHT OF ACID, BASE AND SALT ................................................................. 26
3.4. EQUIVALENT WEIGHT OF AN OXIDANT ............................................................................................ 26
3.5. ATOMIC WEIGHT AND ATOMIC MASS UNIT ..................................................................................... 27
3.6. MOLECULAR WEIGHT........................................................................................................................ 27
3.7. GRAM MOLECULE OR GRAM MOLE.................................................................................................. 28
3.7.1. Molar Volume ................................................................................................................................. 28
i
3.7.2. Mole-Concept ................................................................................................................................. 28
3.8. MASS AND WEIGHT ............................................................................................................................ 29
3.9. AVOGADRO’S HYPOTHESIS AND AVOGADRO’S NUMBER ................................................................. 29
3.9.1. Moles and Molar Solutions (unit = M = moles/L) ......................................................................... 29
4. PREPARATION OF STANDARD SOLUTIONS AND STANDARDIZATION ................................. 32
5. DILUTION OF SOLUTIONS AND DILUTION METHODS ............................................................... 41
6. VOLUMETRIC ANALYSIS ..................................................................................................................... 46
6.2.2 Theory of Acid Base Titration ........................................................................................................ 48
7. INDICATORS ............................................................................................................................................. 51
8. REFERENCES ........................................................................................................................................... 57
List of Tables
Table 1 Recommended Pretreatments of Primary Standards before using them 34
Table 2: Acid base indicators 56
List of Figures
Figure 1 Flow diagram showing the steps in a quantitative analysis ......................................... 2
Figure 2 The relationship between analytical chemistry, other branches of chemistry ............. 5
ii
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Fundamental Analytical Chemistry and Solution Preparation Rev. No: 01
1. Introduction to Analytical Chemistry
“Analytical chemistry is what analytical chemists do.”

“Unless our knowledge is measured and expressed in numbers, it does not amount to much.”
- Lord Kelvin
Analytical chemistry is concerned with the chemical characterization of matter and the answer
to two important questions: what is it (qualitative analysis) and how much is it (quantitative
analysis). Chemicals make up everything we use or consume, and knowledge of the chemical
composition of many substances is important in our daily lives.
Analytical Chemistry provides the methods and tools needed for insight into our material world
. . . for answering four basic questions about a material sample:
✓ What? ✓ What arrangement, structure or
✓ Where? form?
✓ How much?
These covers qualitative, spatial, quantitative, and speciation aspects of analytical science.
1.1. The Nature of Analytical Chemistry
Analytical chemistry is a measurement science consisting of a set of powerful ideas and

methods that are useful in all fields of science and medicine.” Analytical chemistry is
separating, identifying and determining the relative amount of the components in samples.
Qualitative analysis reveals identity of the elements and compounds in a sample while
quantitative analysis indicates the amount of each substance in a sample. The analysis extent
can be complete, elemental or partial based on the intended purpose of the analysis.
There are different analysis methods based on sample kinds and the nature of analytes in
samples:
✓ Wet Chemistry (Classical)
▪ Gravimetric
• ‘Last’ measurement is a mass
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▪ Volumetric
• ‘Last’ measurement is a volume
✓ Instrumental Methods
▪ Electroanalytical
• Measurement of electrical property
▪ Spectrophotometric
• Interaction of light and matter
All methods take advantage of ‘differentiating characteristic’:
• What property changes with the presence/ absence of analyte.
• How (mathematically) does this change relate to the amount off the analyte
A quantitative typical analysis approach is as indicated in the following analysis flow diagram:
Figure 1 Flow diagram showing the steps in a quantitative analysis
There are a number of possible paths through the steps in a quantitative analysis. In the simplest

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example represented by the central vertical pathway, we select a method, acquire and process
the sample, dissolve the sample in a suitable solvent, measure a property of the analyte, calculate
the results, and estimate the reliability of the results. Depending on the complexity of the sample
and the chosen method, various other pathways may also be necessary. Each step in the diagram
explained as:
a) Choosing a method
✓ Accuracy desired
✓ Economic factors
✓ Complexity of the sample and the number of components in the sample
b) Acquiring the sample
✓ Assay: the process of determining how much of a given sample is the material indicated
by its name.
✓ Sampling: the process of collecting a small mass of a material whose composition
accurately represents the bulk of the material being sampled.
c) Processing the sample
✓ Preparing laboratory samples
✓ Defining replicate samples
✓ Preparing solutions: physical and chemical changes
d) Eliminating interferences
✓ Specific method: measure the desired substance accurately in the presence of any
possible combination of foreign substances.
✓ Selective method: determine any of a small group of ion or compounds in the presence
of certain foreign ions or compounds.
✓ Separation technique:
▪ Precipitation
▪ Electrodeposition
▪ Solvent extraction
▪ Ion exchange
▪ Chromatography

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e) Calibration and measurement

f) Calculating results
g) Evaluating results by estimating their reliability
1.2. The Role of Analytical Chemistry
Analytical chemists use science and technology to solve practical problems. Analytical
chemistry is applied in all areas of science, industry, and medicine.
✓ The concentrations of O2 and of CO2 in blood samples.
✓ Quantities of hydrocarbons, NOx, and CO in automobile exhaust gases for emission-
control devices.
✓ Quantitative measurements of ionized Ca in blood serum help diagnose parathyroid
disease in humans.
✓ Quantitative determination of N in foods: protein content and thus their nutritional
value.
✓ Analysis of steel during its production for carbon, nickel, and chromium to achieve a
desired strength, hardness, corrosion resistance, and ductility.
✓ The mercaptan content of household gas supplies to warn of dangerous leaks.
✓ Farmers tailor fertilization and irrigation schedules to meet changing plant needs during
the growing to state few areas in which analytical chemistry is applicable.

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Figure 2 The relationship between analytical chemistry, other branches of chemistry
The central location of analytical chemistry in the diagram signifies its importance and the
breadth of its interactions with many other disciplines.
1.3. Classification of Chemical Analysis
There are a very large number of techniques used in chemical analysis. It can be very useful to
classify the measurement process according to a variety of criteria:
✓ By the type of analytical technique – classical or instrumental techniques;
✓ By the nature of the measurement data generated – single-channel or multi-channel
techniques; and
✓ By the quantitation method (by which the analyte concentration is calculated) – relative
or absolute techniques.

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Classical Vs. Instrumental Techniques
In classical analysis, the signal depends on the chemical properties of the sample: a reagent
reacts completely with the analyte, and the relationship between the measured signal and the
analyte concentration is determined by chemical stoichiometry. In instrumental analysis, some
physical property of the sample is measured, such as the electrical potential difference between
two electrodes immersed in a solution of the sample, or the ability of the sample to absorb light.
Classical methods are most useful for accurate and precise measurements of analyte
concentrations at 0.1% level or higher. On the other hand, some specialized instrumental
techniques are capable of detecting individual atoms or molecules in a sample! Analysis at the
ppm (μg/mL) and even ppb (ng/mL) level is routine.
The advantages of instrumental methods over classical methods include:
✓ The ability to perform trace analysis, as we have mentioned.
✓ Generally, large numbers of samples may be analyzed very quickly.
✓ Many instrumental methods can be automated.
✓ Most instrumental methods are multi-channel techniques.
✓ Less skill and training are usually required to perform instrumental analysis than
classical analysis.
Because of these advantages, instrumental methods of analysis have revolutionized the field of
analytical chemistry, as well as many other scientific fields. However, they have not entirely
supplanted classical analytical methods, due to the fact that the latter are generally more
accurate and precise, and more suitable for the analysis of the major constituents of a chemical
sample. In addition, the cost of many analytical instruments can be quite high. Instrumental
analysis can be further classified according to the principles by which the measurement signal
is generated. A few of the methods are listed below.
Electrochemical methods of analysis, in which the analyte participates in a redox reaction or
other process. In potentiometric analysis, the analyte is part of a galvanic cell, which generates
a voltage due to a drive to thermodynamic equilibrium. The magnitude of the voltage generated
by the galvanic cell depends on the concentration of analyte in the sample solution. In
voltammetry analysis, the analyte is part of an electrolytic cell. Current flows when voltage is

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applied to the cell due to the participation of the analyte in a redox reaction; the conditions of
the electrolytic cell are such that the magnitude of the current is directly proportional to the
concentration of analyte in the sample solution.
Spectrochemical methods of analysis, in which the analyte interacts with electromagnetic
radiation. Most of the methods in this category are based on the measurement of the amount of
light absorbed by a sample; such absorption-based techniques include atomic absorption,
molecular absorption, and NMR methods. The rest of the methods are generally based on the
measurement of light emitted or scattered by a sample; these emission-based techniques include
atomic emission, molecular fluorescence, and Raman scatter methods.
The technique of mass spectroscopy is a powerful method for analysis in which the analyte is
ionized and subsequently detected. Although in common usage, the term “spectroscopy” is not
really appropriate to describe this method, since electromagnetic radiation is not usually
involved in mass spectroscopy. Perhaps the most important use of mass spectrometers in
quantitative analysis is as a gas or liquid chromatographic detector. A more recent innovation
is the use of an inductively coupled plasma (ICP) as an ion source for a mass spectrometer; this
combination (ICP-MS) is a powerful tool for elemental analysis.
Although they do not actually generate a signal in and of themselves, some of the more
sophisticated separation techniques are usually considered “instrumental methods.” These
techniques include chromatography and electrophoresis. These techniques will separate a
chemical sample into its individual components, which are then typically detected by one of the
methods listed above.
Finally, we should note that a number of methods that are based on stoichiometry, and so must
be considered “classical,” still have a significant “instrumental” aspect to their nature. In
particular, the techniques of electrogravimetry, and potentiostatic and amperostatic coulometry
are relatively sophisticated classical methods that have a significant instrumental component.
And let us not forget that instrumental methods can be used for endpoint detection in titrimetric
analysis. Even though potentiostatic titrimetry uses an instrumental method of endpoint
detection, it is still considered a classical method.

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Single-Channel Vs. Multi-Channel Techniques
So now we have classified analytical methods according to the method by which they generate
the measurement data. Another useful distinction between analytical techniques is based on the
information content of the data generated by the analysis:
Single-channel techniques will generate a single number for each analysis of the sample.
Examples include gravimetric and potentiometric analysis. In the former, the signal is a single
mass measurement (e.g., mass of the precipitate) and in the latter method the signal is a single
voltage value.
Multi-channel techniques will generate a series of numbers for a single analysis. Multi-channel
techniques are characterized by the ability to obtain measurements while changing some
independently controllable parameter. For example, in a molecular absorption method, an
absorption spectrum may be generated, in which the absorbance of a sample is monitored as a
function of the wavelength of the light transmitted through the sample. Measurement of the
sample thus produces a series of absorbance values.
Any multi-channel technique can thus produce a plot of some type when analyzing a single
sample, where the signal is observed as a function of some other variable: absorbance as a
function of wavelength (in molecular absorbance spectroscopy), electrode potential as a
function of added titrant volume (potentiometric titrimetry), diffusion current as a function of
applied potential (voltammetry), etc. Multi-channel methods provide a lot more data – and
information – than single-channel techniques.
Multi-channel methods have two important advantages over their single-channel counterparts:
1. They provide the ability to perform multicomponent analysis. In other words, the
concentrations of more than one analyte in a single sample may be determined.
2. Multi-channel methods can detect, and sometimes correct for, the presence of a number of
types of interferences in the sample. If uncorrected, the presence of the interference will result
in biased estimates of analyte concentration.

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1.4. Methods of Analysis
1.4.1. Gravimetric Methods of Analysis
Gravimetry encompasses all techniques in which we measure mass or a change in mass. Mass
is used as a signal in gravimetric analysis. When we step on a scale after exercising, we are
making, in a sense, a gravimetric determination of our mass. Measuring mass is the most
fundamental of all analytical measurements, and gravimetry is unquestionably the oldest
analytical technique.
Types of Gravimetric Methods
• Precipitation gravimetry: A gravimetric method in which the signal is the mass of a

precipitate.
• Electrogravimetry: A gravimetric method in which the signal is the mass of an
electrodeposit on the cathode or anode in an electrochemical cell.
• Volatilization gravimetry: A gravimetric method in which the loss of a volatile species
gives rise to the signal.
• Particulate gravimetry: A gravimetric method in which the mass of a particulate analyte
is determined following its separation from its matrix.
1.4.2. Titrimetric Methods of Analysis
Titrimetry is any method in which volume is the signal. Titrant in titrimetric analysis is the
reagent added to a solution containing the analyte and whose volume is the signal. Equivalence
point in titrimetric analysis is the point in a titration where stoichiometrically equivalent
amounts of analyte and titrant react. End point is the point in a titration where we stop adding
titrant. Indicator is a colored compound whose change in color signals the end point of a
titration. Titration error is the determinate error in a titration due to the difference between the
end point and the equivalence point.
Titrimetric, in which we measure the volume of a reagent reacting stoichiometrically with the
analyte, first appeared as an analytical method in the early eighteenth century. Unlike

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gravimetry, titrimetry initially did not receive wide acceptance as an analytical technique. Many
prominent late-nineteenth century analytical chemists preferred gravimetry over titrimetry and
few of the standard texts from that era include titrimetric methods. By the early twentieth
century, however, titrimetry began to replace gravimetry as the most commonly used analytical
method.
Unlike gravimetry, the growth and acceptance of titrimetry required a deeper understanding of
stoichiometry, thermodynamics, and chemical equilibria. By the early twentieth century the
accuracy and precision of titrimetric methods were comparable to that of gravimetry,
establishing titrimetry as an accepted analytical technique.
Types of Titrimetric Methods
Titrimetric methods are classified into four groups based on the type of reaction involved. These
groups are acid–base titrations, in which an acidic or basic titrant reacts with an analyte that
is a base or an acid; complexometric titrations involving a metal–ligand complexation
reaction; redox titrations, where the titrant is an oxidizing or reducing agent; and precipitation
titrations, in which the analyte and titrant react to form a precipitate.
• Acid–base titration is a titration in which the reaction between the analyte and titrant
is an acid–base reaction.
• Complexation titration is a titration in which the reaction between the analyte and
titrant is a complexation reaction.
• Redox titration is a titration in which the reaction between the analyte and titrant is an
oxidation/reduction reaction.
• Precipitation titration is a titration in which the reaction between the analyte and titrant
involves a precipitation.
1.4.3. Spectroscopic Methods of Analysis
Spectroscopy is a branch of science that studies the interaction between electromagnetic

radiation and matter. Spectrometric methods employ measurement of the intensity of radiation
with a photoelectric transducer or other types of electronic device.

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Spectroscopy Based on Absorption
In absorption spectroscopy a beam of electromagnetic radiation passes through a sample. Much

of the radiation is transmitted without a loss in intensity. At selected frequencies, however, the
radiation’s intensity is attenuated. This process of attenuation is called absorption.
UV-Vis, Infrared & Atomic Absorption instrumental techniques are typical examples of
absorption spectroscopy methods.
Spectroscopy Based on Emission
The release of a photon following thermal excitation is called emission, and that following the
absorption of a photon is called photoluminescence. In chemiluminescence and
bioluminescence, excitation results from a chemical or biochemical reaction, respectively.
Molecular photoluminescence (fluorescence and phosphorescence) and atomic emission
spectroscopy are typical examples of this measurement technique.
Spectroscopy Based on Scattering
The blue color of the sky during the day and the red color of the sun at sunset result from the
scattering of light by small particles of dust, molecules of water, and other gases in the
atmosphere. The efficiency with which light is scattered depends on its wavelength. The sky is
blue because violet and blue light are scattered to a greater extent than other, longer wavelengths
of light. For the same reason, the sun appears to be red when observed at sunset because red
light is less efficiently scattered and, therefore, transmitted to a greater extent than other
wavelengths of light.
Turbidimetry and nephelometry are two related techniques in which an incident source of
radiation is elastically scattered by a suspension of colloidal particles. In turbidimetry, the
detector is placed in line with the radiation source, and the decrease in the radiation’s
transmitted power is measured. In nephelometry, scattered radiation is measured at an angle of
90° to the radiation source.

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1.4.4. Electrochemical Methods of Analysis
Analytical methods in which a measurement of potential, current, or charge in an

electrochemical cell serves as the analytical signal is electrochemical method of analysis.
Potentiometric, coulometric and voltametric methods of analysis are typical examples of this
category of measurement method.
1.4.5. Chromatographic and Electrophoretic Methods of Analysis
Chromatography is a separation in which solutes partition between a mobile and stationary

phase.
In gas chromatography (GC) the sample, which may be a gas or liquid, is injected into a stream
of an inert gaseous mobile phase (often called the carrier gas). The sample is carried through a
packed or capillary column where the sample’s components separate based on their ability to
distribute themselves between the mobile and stationary phases.
In high pressure liquid chromatography (HPLC), a liquid sample, or a solid sample dissolved
in a suitable solvent, is carried through a chromatographic column by a liquid mobile phase.
Separation is determined by solute/stationary-phase interactions, including liquid–solid
adsorption, liquid-liquid partitioning, ion exchange and size exclusion, and by solute/mobile-
phase interactions.
Electrophoresis is another class of separation technique in which analytes are separated based
on their ability to move through a conductive medium, usually an aqueous buffer, in response
to an applied electric field. In the absence of other effects, cations migrate toward the electric
field’s negatively charged cathode, and anions migrate toward the positively charged anode.
More highly charged ions and ions of smaller size, which means they have a higher charge-to-
size ratio, migrate at a faster rate than larger ions, or ions of lower charge. Neutral species do
not experience the electric field and remain stationary. Under normal conditions, even neutral
species and anions migrate toward the cathode. In either case, differences in their rate of
migration allow for the separation of complex mixtures of analytes.

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1.5. Basic Terminologies in Analytical Chemistry
Blank: A blank value is obtained as a result of analysis of a specimen which does not, as far as
possible, contain the analyte (s) in question. Use of various types of blanks (to which no analyte
has been added) enables assessment of how much of the measured instrument response is
attributable to the analyte and how much to other causes. Various types of blank are available to
the user:
i. Reagent blanks: Reagents used during the analytical process (including solvents used for
extraction or dissolution) are analyzed in isolation in order to see whether they contribute
to the measurement signal. The measurement result arising from the analyte can then be
corrected accordingly.
ii. Sample blanks: These are essentially matrices with no analyte. They may be difficult to
obtain but such materials give the best estimate of the effects of interferences that would
be encountered in the analysis of test samples (Eurachem, 1998).
Fortified Sample: An artificially contaminated sample to which a known concentration of
analyte has been added (also known as a spiked sample) which is used to test the accuracy
(especially the efficiency of recovery) of an analytical method.
Recovery: The extraction efficiency of an analytical process, reported as (a percentage of) the
known amount of analyte carried through the sample extraction and processing steps of the
method.
𝐶 𝑠𝑝𝑖𝑘𝑒𝑑 − 𝐶 𝑢𝑛𝑠𝑝𝑖𝑘𝑒𝑑
%𝑅 = 𝑥 100
𝐶 𝑜𝑓 𝑎𝑛𝑎𝑙𝑦𝑡𝑒 𝑠𝑝𝑖𝑘𝑒𝑑
Limit of Detection: A statistical statement about the smallest amount of analyte that can be
determined with confidence.
𝐿𝑂𝐷 = 𝑀𝑒𝑎𝑛 + 3𝑆𝐷
Limit of Quantitation, Limit of Determination: Refers to the smallest analyte concentration or

mass, which can be quantitatively analyzed with a reasonable reliability by a given procedure.

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When an analytical result is below this limit it is reported as less than (<). Another term used is
Reporting limit.
𝐿𝑂𝑄 = 𝑀𝑒𝑎𝑛 + 10𝑆𝐷
Matrix: The predominant material, component or substrate which contains the analyte of
interest.
Analyte: The component of a sample or test item which embodies a quantity or quality that is
ultimately determined directly or indirectly. The term ‘analyte’ is applied to any substance or
material to be analyzed.
Matrix Effects: The direct or indirect alteration or interference in response due to the presence
of unintended analytes (for analysis) or other interfering substances in the sample.
Calibration: Operation that, under specified conditions in a first step, establishes a relation
between the quantity values with measurement uncertainties provided by measurement standards
and corresponding indications with associated measurement uncertainties and, in a second step,
uses this information to establish a relation for obtaining a measurement result from an
indication. A calibration may be expressed by a statement, calibration function, calibration
diagram, calibration curve, or calibration table. In some cases, it may consist of an additive or
multiplicative correction of the indication with associated measurement uncertainty. Calibration
should not be confused with adjustment of a measuring system, often mistakenly called “self-
calibration”, or with verification of calibration (JCGM200:2008).
Traceability: A process whereby the indication of a measuring instrument (or a material
measure) can be compared, in one of more stages, with an international or national standard to
the measurand in question.
Certified Reference Material: Reference material, accompanied by documentation issued by an
authoritative body and providing one or more specified property values with associated
uncertainties and traceability’s, using valid procedures (JCGM200:2008).
Reference Material: Material, sufficiently homogenous and stable with reference to specified
properties, which has been established to be fit for its intended use in measurement of nominal
properties (JCGM200:2008).

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Fitness for Purpose: Degree to which data produced by a measurement process enables a user
to make technically and administratively correct decisions for a stated purpose (IUPAC, 2000).
Method Validation: The process of establishing the performance characteristics and limitations
of a method and the identification of the influences which may change these characteristics and
to what extent.
Which analytes can it determine in which matrices in the presence of which interferences?
Within these conditions what levels of precision and accuracy can be achieved? The process for
verifying that a method is fit for purpose; i.e. for use of solving a particular analytical problem
(Eurachem, 1998)
Repeatability: Condition of measurement, out of a set of conditions that includes the same
measurement procedure, same operators, same measuring system, same operating conditions and
same location, and replicate measurements on the same or similar objects over a short period of
time. Measurement precision under a set of repeatability conditions of measurement
(JCGM200:2008).
Reproducibility: Condition of measurement, out of a set of conditions that includes different
operators, measuring systems, locations, and replicate measurements on the same or similar
objects (JCGM200:2008).
Precision: Closeness of agreement between indications or measured quantity values obtained by
replicate measurements on the same or similar objects under specified conditions.
Measurement precision is used to define measurement repeatability, intermediate measurement
precision, and measurement reproducibility (JCGM200:2008).
Trueness: Closeness of agreement between the average of an infinite number of replicate
measured quantity values and a reference value (JCGM200:2008).
Trueness needs to be determined by replicate testing of a known standard or benchmarking
against proficiency testing results.
Measurement Uncertainty: Result of the evaluation aimed at characterizing the range within
which the true value of a measurand is estimated to lie, generally with a given likelihood.
NOTE: Uncertainty of measurement comprises, in general, many components. Some of these
components may be estimated on the basis of the statistical distribution of the results of series of

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measurements and can be characterized by experimental standard deviations. Estimates of other

components can only be based on experience or other information.
Measurement uncertainty specified as an upper limit and decided on the basis of the intended
use of measurement results (JCGM200:2008) represents the greatest allowed uncertainty for a
particular usage of that result.
Robustness: Robustness of the method is its ability to remain unaffected by small changes in
parameters that are required to control the performance of the method such as temperature,
humidity, mass, time, etc.
Ruggedness: Evaluation of ruggedness is based on a comparison of the standard deviation
obtained when the method is performed in replicate by different test personnel over a period of
time using different standards/reagents and equipment, where possible.
Stability Studies: The stability of culture media, stock solutions, calibration and QC standards,
reference samples and critical reagents shall be verified throughout the duration of their specified
shelf-lives. The easiest and least invasive approach to this requirement is to establish
conservative shelf-lives and measure the method’s performance using the QC system, the
warning and control limits of which are based on the trueness and precision capability of the
method, over the length of each of the designated shelf-lives. The shelf-lives can then be
lengthened in small increments, if desirable, with continued monitoring.
Assessment of stability will be based on t-tests of the mean and f-tests of the variance of the
means and variances over the prescribed period of each of the designated shelf-lives.
Sensitivity: Sensitivity of a method measures the method’s ability to discriminate between small
differences in the determinant’s quantity or concentration.
Selectivity: Property of a measuring system, used with a specified measurement procedure,
whereby it provides measured quantity values for one or more measurands such that the values
of each measurand are independent of other measurands or other quantities in the phenomenon,
body, or substance being investigated (JCGM200:2008).
Working Range: The range of an analytical procedure is the interval between the upper and
lower concentration (amounts) of analyte in the sample (including these concentrations) for
which it has been demonstrated that the analytical procedure has a suitable level of precision,

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accuracy, and linearity. The working range shall be determined by establishing an experiment
where replicate tests are performed on spiked samples or standards over a wide range of
concentrations or measurable quantities and the trueness, precision and linearity are evaluated
for acceptance against customer or laboratory pre-defined requirements.
Linearity: The linearity of an analytical procedure is its ability (within a given range) to obtain
test results which are directly proportional to the concentration (amount) of analyte in the sample.
If the method is a comparative method the linearity of the calibration curve shall be evaluated
using a 6-point (blank and 5 standards) curve. Acceptability of linearity shall be performed by
regression analysis and least mean square fit. In addition, the analytical response from the curve
shall be tested under reproducibility conditions (measure at least 5 -10 times over a period of
time) and the reproducibility standard deviation shall be used to control the equipment set-up
under normal operating conditions using 2 and 3 control lines and the established correlation
coefficient.
1.6. Error in Chemical Analysis
Error is the collective noun for any departure of the result from the "true" value.
1.6.1. Types of Error
i. Random (Indeterminate) Error

ii. Systematic (Determinate) Error
iii. Gross Errors
1.6.1.1. Random (Indeterminate) Errors
These are errors, usually small, which give rise to a spread of results around the average results.
In other words, they define the repeatability or reproducibility of the procedures. They are
unavoidable due to the fact that every physical measurement has limitation, i.e., some uncertainty
using the utmost of care, the analyst can only obtain a weight to the uncertainty of the balance
or deliver a volume to the uncertainty of the glass pipette.

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E.g., with care, an analyst can measure a 1.0000-gram weight (true value) to an accuracy of
±0.0001 grams where a value of 1.0001 to 0.999 grams would be within the random error of
measurement.
1.6.1.2. Systematic (Determinate) Errors
The term 'bias' is sometimes used when defining and describing a systematic error. The measured
value is described as being biased high or low when a systematic error is present.
Some analysts prefer the term 'determinate' instead of systematic because it is more descriptive
in stating that this type of error can be determined. A systematic error can be estimated, but it
cannot be known with certainty because the true value cannot be known. Systematic errors can
therefore be avoided, i.e., they are determinate.
e.g. A chemical standard that has an assigned value that is different from the true value will
always bias the measurements either high or low.
A dirty glass pipette will always deliver less than the intended volume of liquid are errors that
always have the same magnitude and sign, resulting in a bias of the measured values from the
true value. An example would be a ruler missing the first 1 mm of its length – it will consistently
give lengths that are 1 mm short.
Sources of Systematic Error
✓ Instrument error need frequent calibration - both for apparatus such as volumetric flasks,
burettes etc., but also for electronic devices such as spectrometers.
✓ Method error due to inadequacies in physical or chemical behaviour of reagents or
reactions (e.g. slow or incomplete reactions). For example, nicotinic acid does not react
completely under normal Kjeldahl conditions for nitrogen determination method and an
accepted reference value. The method bias may depend on the analyte level.
✓ Personal error e.g. insensitivity to colour changes; tendency to estimate scale readings
to improve precision; preconceived idea of “true” value.
How the above errors minimised?

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1.6.1.3. Gross Errors
Gross errors are undetected mistakes that cause a measurement to be very much farther from the
mean measurement than other measurements; are errors that are so serious (i.e. large in
magnitude) that they cannot be attributed to either systematic or random errors associated with
the sample, instrument, or procedure. An example would be writing down a value of 100 when
the reading was actually 1.00. If included in calculations, gross errors will tend to affect both
accuracy and precision.
2. Concentration and Methods of Expressing Concentration
Here, units of concentration and how to convert from one concentration unit to the other unit of
concentration is discussed. It will be important to understand a few terms dealing with solutions.
✓ Solution: a homogeneous mixture of one or more substances (the solutes) dissolved in
another substance (solvent).
✓ Solute: component of a solution present in the lesser amount.
✓ Solvent: component of a solution present in greater amount.
✓ Concentration of a Solution: means grams of solute dissolved per liter of solution.
Usually it is expressed in terms of normality and molarity.
Expression of Concentration
2.1. Molarity (M)
Is the number of moles of solute per volume of solution in liter.
𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑚𝑜𝑙𝑒𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒

𝑀𝑜𝑙𝑎𝑟𝑖𝑡𝑦 (𝑀) = 𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 𝑖𝑛 𝑙𝑖𝑡𝑒𝑟
Note:
✓ Number of moles of solute (n) is the mass of the solute taken per its molecular weight.
✓ Molarity is the most common concentration unit involved in calculations dealing with
volumetric stoichiometry.

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2.2. Normality (N)
Is the number of grams of solute divided by equivalent weight of the solute per volume in liter
of solution.
𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓𝑔𝑟𝑎𝑚𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒 𝑖𝑛 1000 𝑚𝑙 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛

𝑁𝑜𝑟𝑚𝑎𝑙𝑖𝑡𝑦 = 𝑒𝑞𝑢𝑖𝑣𝑎𝑙𝑒𝑛𝑡 𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑡ℎ𝑒 𝑠𝑜𝑙𝑢𝑡𝑒
Note: Equivalent weight of solute is molecular weight divided by number of equivalents of the
solute.
2.3. Molality (m)

Is the number of moles of solute per kilogram of solvent.
𝑛𝑢𝑚𝑏𝑒𝑟𝑜𝑓𝑚𝑜𝑙𝑒𝑠𝑜𝑓𝑠𝑜𝑙𝑢𝑡𝑒
𝑀𝑜𝑙𝑎𝑙𝑖𝑡𝑦 =
𝑚𝑎𝑠𝑠 𝑜𝑓𝑠𝑜𝑙𝑣𝑒𝑛𝑡 𝑖𝑛 𝑘𝑖 𝑙𝑜𝑔 𝑟 𝑎𝑚
Note: Molality is often used as the concentration unit involved in calculations dealing with
colligative properties, such as freezing point depression, boiling point elevation and osmotic
pressure.
2.4. Mole-fraction (x)
Is the ratio of the number of moles of one component to the total mole.
𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓𝑚𝑜𝑙𝑒𝑠 𝑜𝑓 𝐴
𝑀𝑜𝑙𝑒 𝑓𝑟𝑎𝑐𝑡𝑖𝑜𝑛 𝑜𝑓𝐴 (𝑥𝐴 ) =
𝑡𝑜𝑡𝑎𝑙 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑚𝑜𝑙𝑒𝑠
2.5. Percentage Concentration
This unit of concentration is often used for concentrated solutions, typically acids and bases. If
you were to look on a bottle of a concentrated acid or base solution the concentration expressed
as a weight/weight percent.

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2.6. Weight/ Weight (w/w %)
Expresses the mass of solute contained in 100 units of mass of solution.
𝑚𝑎𝑠𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒
𝑤 ⁄𝑤 % = 𝑚𝑎𝑠𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 𝑥100%
2.7. Volume/ Volume (v/v %)
Expresses the volume of liquid solute contained in 100 units of volume of solution.
𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒
𝑣/𝑣% = 𝑥100%
𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
2.8. Weight/ Volume (w/v %)
Expresses the mass of solute contained in 100 units of volume of solution.
𝑚𝑎𝑠𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒
𝑤/𝑣% = 𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 𝑥100%
2.9. Parts per Million (ppm)
This unit of concentration may be expressed in a number of ways. It is often used to express the
concentration of very dilute solutions. The “Technical” definition of parts per million is:
𝑔𝑟𝑎𝑚𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒
𝑝𝑝𝑚 = 𝑔𝑟𝑎𝑚𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 𝑥10−6
Since the amount of solute relative to the amount of solvent is typically very small, the density
of the solution is to a first approximation the same as the density of the solvent. For this reason,
parts per million may also be expressed in the following two ways:
𝑚𝑔 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒 𝑚𝑔 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒
𝑝𝑝𝑚 = 𝐿 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 Or 𝑝𝑝𝑚 = 𝐾𝑔 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛

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2.10. Parts per Billion (ppb)
This concentration unit is also used for very dilute solutions. The “technical” definition is as
follows:
𝑔𝑟𝑎𝑚𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒
𝑝𝑝𝑏 = 𝑔𝑟𝑎𝑚𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 𝑥10−9
Owing to the dilute nature of the solution, once again, the density of the solution will be about
the same as the density of the solvent. Thus, we may also express parts per billion as:
𝜇𝑔 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒 𝜇𝑔 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒
𝑝𝑝𝑏 = 𝐿 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 Or 𝑝𝑝𝑏 = 𝐾𝑔 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
2.11. Formal Solution
A formal solution is one in which contains a formula weight of a solute in a liter of the solution.
It is denoted by F. In most of the cases formula weights and molecular weights are identical but
sometimes the true molecular weight of a compound is a multiple of the weight expressed by its
formula as ordinary written in a chemical reaction.
𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑔𝑟𝑎𝑚𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒 𝑝𝑒𝑟 𝑙𝑖𝑡𝑒𝑟 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛

𝐹𝑜𝑟𝑚𝑎𝑙𝑖𝑡𝑦 (𝐹) =
𝑓𝑜𝑟𝑚𝑢𝑙𝑎 𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒
Factor of Solution: The factor of a solution is a number with which the concentration of the
proposed standard solution is to be multiplied to indicate the actual concentration of the prepared
solution. It is not always possible to weigh out exact amount of solute to prepare a solution of
exact concentration. In fact, an amount nearest to the weight required is accurately weighed. For
example, to prepare 1 liter of (N/10) Na2CO3 solution 5.3 g is required. Let the actual weight
taken be 5.45g. Now, 5.3 g of Na2CO3 when present in 1 liter, the concentration is (N/10). 5.45
g of Na2CO3 when present in 1 liter, the concentration is 5.45/5.3(N/10) or 1.028(N/10).
Here 1.028 is the normality factor.
𝑤𝑒𝑖𝑔ℎ𝑡 𝑎𝑐𝑡𝑢𝑎𝑙𝑙𝑦 𝑡𝑎𝑘𝑒𝑛

𝐹𝑎𝑐𝑡𝑜𝑟 𝑜𝑓 𝑎 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 (𝐹) =
𝑤𝑒𝑖𝑔ℎ𝑡 𝑟𝑒𝑞𝑢𝑖𝑟𝑒𝑑 𝑡𝑜 𝑏𝑒 𝑡𝑎𝑘𝑒𝑛

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You may find it necessary to be able to convert from one concentration unit to another. The key
to solving this type of problem is to realize that you may make an assumption to get started. You
may need to know the density of the solution, which would be given in the problem. Then, by
using dimensional analysis, you try to get to the units of the concentration unit you are seeking
to find. To get started, assume the quantity of solution found in the denominator unit of the
concentration unit you are trying to convert. For example, if you are trying to convert
weight/weight percent to molarity, assume 100 grams of solution. If you are trying to convert
molarity to weight/weight percent, assume 1 liter of solution. Let’s look at a typical example.
Suppose you are given a concentrated solution of HCl which is known to be 37.0 % HCl and has
a solution density of 1.19 g/ml. What is the molarity, molality, and mole fraction of HCl?
As stated above, begin with the assumption of 100 g of solution. With this assumption, you now
know a few other facts. In 100 g of solution, 37.0 g is due to HCl (grams of solute) and 63 g is
due to water (grams of solvent).
To find molarity, we need to determine the moles of HCl (solute) per liter of solution. First,
convert the known amount of HCl (37.0 g) to moles:
1 𝑚𝑜𝑙 𝐻𝐶𝑙
𝑚𝑜𝑙 𝐻𝐶𝑙 = 37.0𝑔 𝐻𝐶𝑙 𝑥 = 1.01 𝑚𝑜𝑙 𝐻𝐶𝑙
36.5𝑔 𝐻𝐶𝑙
Next, convert the known mass of solution, 100 g solution, to liters of solution, using the density
of the solution:
1𝑚𝑙 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 1𝐿 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
𝐿 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 = 100𝑔 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 𝑥 𝑥 = 0.0840𝐿 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
1.19𝑔 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 1000𝑚𝐿 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
Since the moles of solute (HCl) and volume of solution in liters is now known, calculate the
molarity (M) as the moles of solute per liter of solution:
1.01𝑚𝑜𝑙 𝐻𝐶𝑙
𝑀= = 12.0 𝑚𝑜𝑙 𝐻𝐶𝑙/𝐿 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
0.0840𝐿 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
From the information above, let’s find the molality of the HCl solution. The mole of solute is
already known (1.01 mol HCL). We need to find the kilogram of solvent:

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1𝐾𝑔 𝐻 𝑂
63.0𝑔𝐻2 𝑂𝑥 1000𝑔 𝐻2 = 0.0630𝐾𝑔 𝐻2 𝑂
2𝑂
Since molality (m) is defined as the moles of solute per kilogram of solvent, it becomes easy to
find the molality:
𝑚 = 0.0630𝐾𝑔 𝐻 = 16.0𝑚𝑜𝑙 𝐻𝐶𝑙/𝐾𝑔 𝐻2 𝑂
2𝑂
Finally, let’s tackle the mole fraction of HCl. The mole of HCl is known to be 1.01 moles. We
need to find the moles of H2O:
1𝑚𝑜𝑙 𝐻2 𝑂
𝑚𝑜𝑙 𝐻2 𝑂 = 63.0𝑔 𝐻2 𝑂𝑥 = 3.50𝑚𝑜𝑙 𝐻2 𝑂
18.0𝑔 𝐻2 𝑂
Since the moles of solute (HCl) and the moles of solvent (H2O) are known, the mole fraction of
HCl may be calculated:
𝑋𝐻𝐶𝑙 = = 0.224
1.01𝑚𝑜𝑙 𝐻𝐶𝑙 + 3.50𝑚𝑜𝑙 𝐻2 𝑂
2.12. Equivalent Weight
The equivalent weight of a substance is defined as the weight of an element or compound which
combines with or displaces from combination 8.00 parts by weight of oxygen, or 1.008 parts by
weight of hydrogen or 35.45 parts by weight of chlorine.
3. Variability in Equivalent Weights
The equivalent weight of an element is decided on the basis of combination in its compounds.
An element having variable valency can combine with an element with single valency to form
more than one compound. As a result, the equivalent weight of the element in the compounds
will be different since the ratio of the weights of the two elements in the compounds is different.
But if the element has only one valency, then it would form only one compound with the second
element i.e. the ratio of the weight of the two elements in the compounds would be fixed. Hence
the equivalent weight of the element will also be fixed. Many elements like Cu, Fe, N, P, etc.

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show more than one valency. The equivalent weight of such elements, therefore, varies from one
compound to the other. For example, iron forms two oxides viz. FeO (ferrous oxide) and Fe2O3
(ferric oxide). In FeO the equivalent weight of iron is 28 and in Fe2O3 it is 18.33.
3.1. Equivalent Weight and Valency
Let V and E be the valency and equivalent weight of the element respectively and A be the
atomic weight. Now one atom of the element will combine with V atoms of hydrogen, since
valency is the number of hydrogen atoms which an atom of the element combines with. The
weight of V atoms of hydrogen is V (since atomic weight of hydrogen = 1).
Now, 1 part by weight of hydrogen combines with A/V parts of the element. Therefore, A/V is
the equivalent weight of the element, i.e. E = A/V or A = E x V, i.e. Atomic weight = Equivalent
Weight x Valency. When V = 1, A = E, i.e. for monovalent elements atomic weight is the same
as the equivalent weight.
3.2. Equivalent Weight of Acid, Base and Salt
The equivalent weight of an acid is the number of parts by weight of the acid containing one part
by weight of replaceable hydrogen. Hence,
𝑀𝑊 𝑜𝑓 𝑡ℎ𝑒 𝐴𝑐𝑖𝑑
𝐸𝑞. 𝑤𝑡 𝑜𝑓𝐴𝑐𝑖𝑑 =
𝑁𝑜 𝑜𝑓 𝑟𝑒𝑝𝑙𝑎𝑐𝑒𝑎𝑏𝑙𝑒 𝐻 𝑎𝑡𝑜𝑚𝑠 𝑖𝑛 𝑜𝑛𝑒 𝑚𝑜𝑙. 𝑜𝑓 𝑡ℎ𝑒 𝐴𝑐𝑖𝑑
𝑀𝑊 𝑜𝑓 𝑡ℎ𝑒 𝐴𝑐𝑖𝑑
=
𝐵𝑎𝑠𝑖𝑐𝑖𝑡𝑦 𝑜𝑓 𝑡ℎ𝑒 𝐴𝑐𝑖𝑑
The equivalent weight of a base may be defined as the number of parts by weight of the base that
neutralizes one equivalent of an acid.
𝑀𝑊 𝑜𝑓 𝑡ℎ𝑒 𝐵𝑎𝑠𝑒
𝐸𝑞. 𝑤𝑡 𝑜𝑓 𝑎 𝐵𝑎𝑠𝑒 =
𝑁𝑜 𝑜𝑓 𝑟𝑒𝑝𝑙𝑎𝑐𝑒𝑎𝑏𝑙𝑒 𝑂𝐻 − 𝑔𝑟𝑜𝑢𝑝𝑠 𝑖𝑛 𝑜𝑛𝑒 𝑚𝑜𝑙. 𝑜𝑓 𝑡ℎ𝑒 𝐵𝑎𝑠𝑒
𝑀𝑊 𝑜𝑓 𝑡ℎ𝑒 𝐵𝑎𝑠𝑒
=
𝐴𝑐𝑖𝑑𝑖𝑡𝑦 𝑜𝑓 𝑡ℎ𝑒 𝐵𝑎𝑠𝑒
The equivalent weight of a salt may be defined as the number of parts by weight of the salt
containing one equivalent part of the metal present in the salt. Thus, equivalent weight of a salt

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may be obtained by dividing the molecular weight of the salt by the total valency of the metal
atom or atoms present in one molecule of the salt. Hence,
𝑀𝑊𝑜𝑓𝑡ℎ𝑒 𝑆𝑎𝑙𝑡
𝐸𝑞. 𝑤𝑡 𝑜𝑓𝑎 𝑠𝑎𝑙𝑡 =
𝑁𝑜 𝑜𝑓 𝑚𝑒𝑡𝑎𝑙 𝑎𝑡𝑜𝑚𝑠 𝑖𝑛 𝑜𝑛𝑒 𝑚𝑜𝑙. 𝑜𝑓 𝑡ℎ𝑒 𝑆𝑎𝑙𝑡 𝑥 𝑣𝑎𝑙𝑒𝑛𝑐𝑦 𝑜𝑓 𝑡ℎ𝑒 𝑚𝑒𝑡𝑎𝑙 𝑎𝑡𝑜𝑚 (𝑠)
3.3. Gram-Equivalent Weight of Acid, Base and Salt
𝐺𝑟𝑎𝑚 𝑀𝑊 𝑜𝑓 𝑡ℎ𝑒 𝐴𝑐𝑖𝑑

𝐺𝑟𝑎𝑚 𝐸𝑞. 𝑊𝑡 𝑜𝑓 𝑎𝑛 𝐴𝑐𝑖𝑑 =
𝐵𝑎𝑠𝑖𝑐𝑖𝑡𝑦 𝑜𝑓 𝑡ℎ𝑒 𝐴𝑐𝑖𝑑
𝐺𝑟𝑎𝑚 𝑀𝑊𝑜𝑓𝑡ℎ𝑒 𝐵𝑎𝑠𝑒

𝐺𝑟𝑎𝑚 𝐸𝑞. 𝑊𝑡 𝑜𝑓𝑎 𝐵𝑎𝑠𝑒 =
𝐴𝑐𝑖𝑑𝑖𝑡𝑦 𝑜𝑓 𝑡ℎ𝑒 𝐵𝑎𝑠𝑒
𝐺𝑟𝑎𝑚 𝑀𝑊 𝑜𝑓 𝑡ℎ𝑒 𝑆𝑎𝑙𝑡

𝐺𝑟𝑎𝑚 𝐸𝑞. 𝑊𝑡 𝑜𝑓 𝑎 𝑆𝑎𝑙𝑡 =
𝑇𝑜𝑡𝑎𝑙 𝑝𝑜𝑠𝑖𝑡𝑖𝑣𝑒 𝑣𝑎𝑙𝑒𝑛𝑐𝑦 𝑖𝑛 𝑡ℎ𝑒 𝑓𝑜𝑟𝑚𝑢𝑙𝑎 𝑜𝑓 𝑡ℎ𝑒 𝑠𝑎𝑙𝑡
3.4. Equivalent Weight of an Oxidant
The equivalent weight of an oxidant or reductant is the mole divided by the number of electrons
which one mole of the substance gains or losses in the reaction.
For example,
− + − 2+
𝑀𝑛𝑂4 − 𝐾𝑀𝑛𝑂4
𝑀𝑛𝑂4 + 8𝐻 + 5𝑒 → 𝑀𝑛 + 4𝐻2 𝑂; 𝐸𝑞. = =
5 5
2−
𝐶𝑟 𝑂
2 7 𝐾2 𝐶𝑟2 𝑂7
𝐶𝑟2 𝑂7 2− ⥂ +14𝐻 + + 6𝑒 − → 2𝐶𝑟 3+ + 7𝐻2 𝑂; 𝐸𝑞. = =
6 6
2+
𝐹𝑒 𝐹𝑒𝑆𝑂 4
𝐹𝑒 2+ → 𝐹𝑒 3+ + 𝑒 − ; 𝐸𝑞. = =
1 1
2−
𝐶2 𝑂4 𝐻2 𝐶2 𝑂4
𝐶2 𝑂4 2− → 2𝐶𝑂2 + 2𝑒 − ; 𝐸𝑞. = =
2 2
𝑆𝑂3 2− 𝑁𝑎2 𝑆𝑂3
𝑆𝑂3 2− + 𝐻2 𝑂 → 𝑆𝑂4 2− + 2𝐻 + + 2𝑒 − ; 𝐸𝑞. = =
2 2
2−
𝑆 𝑂
2 3 𝑁𝑎 𝑆 𝑂
2 2 3
2𝑆2 𝑂3 2− → 𝑆4 𝑂6 + 2𝑒 − ; 𝐸𝑞. = =
1 1
Milliequivalents per Liter
A solution containing milligram equivalent (1/1000 g equivalent) of a substance in a liter of

solution is expressed as meq/liter.

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3.5. Atomic Weight and Atomic Mass Unit
The atomic weight of an element may be defined as a number that gives the mass of one atom
of that element compared with mass of one atom of oxygen taken as exactly 16 or mass of one
atom of carbon taken as exactly 12.
𝑚𝑎𝑠𝑠 𝑜𝑓𝑜𝑛𝑒 𝑎𝑡𝑜𝑚 𝑜𝑓 𝑡ℎ𝑒 𝑒𝑙𝑒𝑚𝑒𝑛𝑡

𝐴𝑡. 𝑊𝑡 = 𝑥 16
𝑚𝑎𝑠𝑠 𝑜𝑓 𝑜𝑛𝑒 𝑎𝑡𝑜𝑚 𝑜𝑓 𝑜𝑥𝑦𝑔𝑒𝑛
𝑚𝑎𝑠𝑠 𝑜𝑓𝑜𝑛𝑒 𝑎𝑡𝑜𝑚 𝑜𝑓 𝑡ℎ𝑒 𝑒𝑙𝑒𝑚𝑒𝑛𝑡

= 𝑥 12
𝑚𝑎𝑠𝑠 𝑜𝑓 𝑜𝑛𝑒 𝑎𝑡𝑜𝑚 𝑜𝑓 𝑐𝑎𝑟𝑏𝑜𝑛
The atomic mass unit (a.m.u) is the unit by which atomic weight is expressed.
1 a.m.u = 1/16 x mass of one oxygen atom.

= 1/12 x mass of one carbon atom. Therefore, atomic weight of oxygen = 16 a.m.u.
A gram atom of an element is the quantity of the element, the weight of which in grams is
numerically equal to the atomic weight of the element. Thus, a gram atom of oxygen is 16 grams.
3.6. Molecular Weight
The molecular weight of a substance is a number that gives the weight of one molecule of that
substance compared with the weight of one atom of oxygen taken as exactly 16 (or one atom of
carbon taken as exactly 12).
𝑊𝑡 𝑜𝑓 𝑜𝑛𝑒 𝑚𝑜𝑙𝑒𝑐𝑢𝑙𝑒 𝑜𝑓 𝑡ℎ𝑒 𝑠𝑢𝑏𝑠 𝑡𝑎𝑛 𝑐 𝑒

𝑀𝑊 = 𝑥 16
𝑊𝑡 𝑜𝑓 𝑜𝑛𝑒 𝑎𝑡𝑜𝑚 𝑜𝑓 𝑐𝑎𝑟𝑏𝑜𝑛
𝑊𝑡 𝑜𝑓 𝑜𝑛𝑒 𝑚𝑜𝑙𝑒𝑐𝑢𝑙𝑒 𝑜𝑓 𝑡ℎ𝑒 𝑠𝑢𝑏𝑠 𝑡𝑎𝑛 𝑐 𝑒

= 𝑥 12
𝑤𝑡 𝑜𝑓 𝑜𝑛𝑒 𝑎𝑡𝑜𝑚 𝑜𝑓 𝑐𝑎𝑟𝑏𝑜𝑛
Molecular weight of a substance can be obtained by adding atomic weights of the various atoms
of which a molecule is composed. Thus, the molecular weight of chlorine from its formula Cl2
is weight of 2Cl atoms
= 2 x 35.5 = 71.

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3.7. Gram Molecule or Gram Mole
The weight of a substance in grams which is equal to its molecular weight is known as gram-
molecule or gram mole. The molecular weight of water is 18. Therefore, one-gram mole of water
weighs 18 grams.
3.7.1. Molar Volume
A gram mole of any gas at STP, i.e. standard temperature and pressure (273 K and 760 mm Hg),
occupies a volume of 22.4 liters. This is the molar volume. Thus 2 gram of hydrogen gas or 32
gram of oxygen gas will occupy a volume of 22.4 liters at STP.
3.7.2. Mole-Concept
Gram molecule of any substance is considered as mole. Both gram molecule and gram atom can
be represented by mole. The term ‘mole’ of a substance may be defined as the weight in grams
which contain 6.023 x 1023 molecules. In the case of mono atomic molecule one mole represents
the weight in grams of the element which contains 6.023 x 1023 atoms of the element.
In chemical analysis and calculation, the weights of reacting substances are considered through
the molecules and the atoms of the substances taking part in the chemical reaction. The term
‘mole’ indicates at the same time the weight of the reacting substances as well as the number of
molecules present in them. Here lies the significance of mole.
The term mole may be considered as follows:
✓ 1 mole = gram-molecule = molecular weight in gram.
✓ For gases, 1 mole = 22.4 liters at STP.
✓ 1 mole = 6.023 x 1023 molecules or atoms = Avogadro’s number.
Besides, mole is also used to represent one-gram ion and one Faraday of electricity. One mole
of oxygen is the amount of oxygen in grams which contains Avogadro’s number of oxygen
atoms, i.e. 6.023 x 1023 atoms. One mole of oxygen is the molecular weight of oxygen expressed
in grams (i.e. 16 grams of oxygen).
One mole of nitrogen molecules means one-gram molecule of nitrogen or 28 grams of nitrogen
or 6.023x1023 molecules of nitrogen.

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One mole of ammonium ions means one-gram ion of ammonium ions or 18 grams of ammonium
ions or 6.023 x 1023 ammonium ions.
3.8. Mass and Weight
The ‘mass’ of a substance is a definite property which can be used as a measure of quantity. The
‘weight’ of the substance is the force which results from the interaction of the gravitational force
on the substance. Weight is thus the product of mass and acceleration due to gravity.
3.9. Avogadro’s Hypothesis and Avogadro’s Number
Avogadro’s hypothesis (1811) states “equal volumes of all gases under STP contain equal
number of molecules”. Thus, according to this hypothesis if one liter of hydrogen gas contains
‘n’ molecules at 0oC and 760 mm Hg pressure, one liter of any other gas (e.g. ammonia, chlorine,
oxygen, nitrogen, carbon dioxide, etc.) would contain ‘n’ molecules under the same conditions
of temperature and pressure. Experimental results have verified the above stated fact.
Avogadro’s number is the number of molecules present in one gram-molecule of a substance.
One-gram molecule of any gas at STP occupies a volume of 22.4 liters. So, Avogadro’s number
also denotes the number of molecules present in 22.4 liters of any gas at 0oC and 760 mm Hg
pressure. This number is a constant and its value is 6.023 x 1023. This means that molecular
weight of any substance expressed in grams contains 6.023 x 1023 atoms. Thus 1 gram-molecule
hydrogen (2g) and 1-gram molecule oxygen (32 g) though differ in weight, each of them contains
equal number of molecules i.e. 6.023 x 1023.
3.9.1. Moles and Molar Solutions (unit = M = moles/L)
Sometimes it may be more efficient to use molarity when calculating concentrations. A mole is
defined as one-gram molecular weight of an element or compound, and comprised of exactly
6.023 x 1023 atoms or molecules (this is called Avogadro’s number). The mole is therefore a unit
expressing the amount of a chemical. The mass (g) of one mole of an element is called its
molecular weight (MW). When working with compounds, the mass of one mole of the
compound is called the formula weight (FW). The distinction between MW and FW is not

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always simple, however, and the terms are routinely used interchangeably in practice. Formula
(or molecular) weight is always given as part of the information on the label of a chemical bottle.
The number of moles in an arbitrary mass of a dry reagent can be calculated as:
𝑁𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑚𝑜𝑙𝑒𝑠 = 𝑤𝑒𝑖𝑔ℎ𝑡 (𝑔)/ 𝑚𝑜𝑙𝑒𝑐𝑢𝑙𝑎𝑟 𝑤𝑒𝑖𝑔ℎ𝑡 (𝑔)
Molarity is the unit used to describe the number of moles of a chemical or compounds in one
liter (L) of solution and is thus a unit of concentration. By this definition, a 1.0 Molar (1.0 M)
solution is equivalent to one formula weight (FW = g/mole) of a compound dissolved in 1 liter
(1.0 L) of solvent (usually water).
Example 1: To prepare a liter of a simple molar solution from a dry reagent:
Multiply the formula weight (or MW) by the desired molarity to determine how many grams of
reagent to use:
Chemical FW = 194.3 g/mole; to make 0.15 M solution use
194.3 g/mole x 0.15 moles/L = 29.145 g/L
Example 2: To prepare a specific volume of a specific molar solution from a dry reagent:
A chemical has a FW of 180 g/mole and you need 25 ml (0.025 L) of 0.15 M (M = moles/L)
solution. How many grams of the chemical must be dissolved in 25 ml water to make this
solution?
#grams/desired volume (L) = desired molarity (mole/L) x FW (g/mole)
By algebraic rearrangement,
#grams = desired volume (L) x desired molarity (mole/L) x FW (g/mole)
#grams = 0.025 L x 0.15 mole/L x 180 g/mole
After canceling the units,
#grams = 0.675 g so, you need 0.675 g/25 ml

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Percent Solutions (% = parts per hundred or grams/100 ml)
Many reagents are mixed as percent concentrations as weight per volume for dry reagent OR
volume per volume for solutions. When working with a dry reagent it is mixed as dry mass (g)
per volume and can be simply calculated as the % concentration (expressed as a proportion or
ratio) x volume needed = mass of reagent to use.
Example 1: If you want to make 200 ml of 3 % NaCl you would dissolve 0.03 g/ml x 200 ml =
6.0 g NaCl in 200 ml water.
When using liquid reagents, the percent concentration is based upon volume per volume, and is
similarly calculated as % concentration x volume needed = volume of reagent to use.
Example 2: If you want to make 2 L of 70% acetone you would mix 0.70 ml/ml x 2000 ml =
1400 ml acetone with 600 ml water.
To convert from % solution to molarity, multiply the % solution by 10 to express the percent
solution grams/L, then divides by the formula weight.
𝑔𝑟𝑎𝑚𝑠 𝑜𝑓 𝑟𝑒𝑎𝑔𝑒𝑛𝑡⁄
( 100 𝑚𝑙 )
𝑀𝑜𝑙𝑎𝑟𝑖𝑡𝑦 = × 10
𝐹𝑊
Example 3: Convert a 6.5 % solution of a chemical with FW = 325.6 to molarity,
[(6.5 g/100 ml) x 10] / 325.6 g/mole = [65 g/L] / 325.6g/mole = 0.1996 M
To convert from molarity to percent solution, multiply the molarity by the FW and divide by 10:
𝑚𝑜𝑙𝑎𝑟𝑖𝑡𝑦 × 𝐹𝑊
%𝑆𝑜𝑙𝑢𝑡𝑖𝑜𝑛 =
10
Example 4: Convert a 0.0045 M solution of a chemical having FW 178.7 to percent solution:

[0.0045 moles/L x 178.7 g/mole] / 10 = 0.08 % solution.
Concentrated Stock Solutions - Using "X" Units

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Stock solutions of stable compounds are routinely maintained in labs as more concentrated
solutions that can be diluted to working concentrations when used in typical applications. The
usual working concentration is denoted as 1X. A solution 20 times more concentrated would be
denoted as 20X and would require a 1:20 dilution to restore the typical working concentration.
Example 5: A 1X solution of a compound has a molar concentration of 0.05 M for its typical
use in a lab procedure. A 20X stock would be prepared at a concentration of 20 x 0.05 M = 1.0
M. A 30X stock would be 30 x 0.05 M = 1.5 M.
Normality (N): Conversion to Molarity
𝑁𝑜𝑟𝑚𝑎𝑙𝑖𝑡𝑦 = 𝑛 𝑥 𝑀
Where, n = number of protons (H+) in a molecule of the acid.
Example 6: In the formula for concentrated sulphuric (36 N H2SO4), there are two protons, so,
its molarity = N/2. So, 36N H2SO4 = 36/2 = 18 M.
4. Preparation of Standard Solutions and Standardization
Standard solutions are solutions of accurately known concentration. There are two methods for
preparing standard solutions.
1. A quantity of the required substance is weighed out accurately on an analytical balance,
dissolved in a measuring (volumetric) flask, and the volume is made up to the mark with water.
If the weight of dissolved substance (g) and the volume of solution (V) are known, it is easy to
calculate the titer of the solution. This is evidently:
𝑔 𝑔
𝑇 = 𝑉 ( ⁄𝑚𝑙 )
Some primary standard solutions are prepared from their respective pure solid crystals by the
stated way. Obviously, this method of preparing standard solutions is very often not applicable
for some others. It cannot be used for preparing standard solutions of such substances as HCl,
NaOH, etc. In fact, the exact concentration of an aqueous HCl solution is never known.

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Therefore, even if an exact weight of such a solution is taken, it is impossible to calculate the
weight of hydrogen chloride it contains. The same applies to NaOH, which readily absorbs CO2
and water vapor from the air, with a change of weight. Therefore, the amount of NaOH in a
weighed sample is not known exactly either.
These examples show that the method described above can be used for preparation of standard
solutions only of substances which satisfy the following conditions:
✓ The substance must be chemically pure, i.e., it must not contain impurities in amounts
which could affect analytical precision (not over 0.05 – 0.1 %).
✓ The substance must be stable on keeping; both in the solid state and in solution, as
otherwise its composition would cease to correspond to its formula.
✓ The gram-equivalent of the substance should be as large as possible, so that the precision
in determining the normality of the solution is high.
Substances satisfying these conditions are known as primary, as all other substances are
standardized against them.
2. If the substance does not satisfy all the above conditions, a solution of approximately the
required normality is first prepared. At the same time a standard solution of some suitable
primary substance is prepared as described above. One of these solutions is then titrated with the
other, and the exact concentration (titre) of the approximate solution is calculated, the
concentration of the primary standard solution being known.
For example, the titer of a NaOH solution can be found by titrating a solution of oxalic acid with
it. Oxalic acid is a crystalline substance which can be obtained chemically pure by
recrystallization so that it exactly corresponds to its formula H2C2O4.2H2O. The titre of the oxalic
acid solution can therefore be found by dividing the exact weight of oxalic acid taken by the
solution volume.
A standard solution, the titer of which is found by titration (as in the case of NaOH) or by
gravimetric analysis of the solution is described as standardized.
The great importance of primary substances in volumetric analysis is evident from all this. The
better they satisfy the above conditions, the more accurately can the titers of the “working”
solutions used in analysis be found, and less will the analytical errors be.

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It should be noted that working solutions are not always standardized against primary standards.
For example, the titer of a NaOH solution can also be found by titration with HCl solution which
has been standardized against a suitable primary standard. This method is convenient in that
fewer primary standards are needed and therefore the time required for their purification is saved.
However, it is less accurate because the errors in determinations of the separate titers are
cumulative.
Solutions are sometimes standardized by gravimetric analysis. For example, the titer of an HCl
solution can be found from the weight of the AgCl precipitate formed by the action of AgNO3
solution on a known weight of the HCl solution. Similarly, to find the titer of an H2SO4 solution
we can add BaCl2 solution to a definite volume of it and weigh the BaSO4 precipitate formed,
etc.
The acids generally used are hydrochloric acid and sulfuric acid. Hydrochloric acid is preferred
in the preparation of standard acid since all the chlorides are soluble in water. All the sulfates
are not easily soluble. The sulfates of calcite and barium are very sparingly soluble in water. This
section provides some information on how to prepare standard solutions.
Table 1 Recommended Pretreatments of Primary Standards before using them
Recommended Pretreatments
S/N Primary Standards Temperature, oC Time, hour
1 Calcium carbonate 105 2
2 Sodium carbonate 270 ± 10 2
3 Sodium chloride ~ 200 24
4 Sodium oxalate 105 2
5 Potassium hydrogen phthalate 105 2
6 Potassium iodate 180 3
7 Arsenic trioxide (poison) 105 – 110 3
8 Potassium dichromate 150 - 200 3

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Examples:
Preparation of 0.1N Hydrochloric Acid (HCl)
HCl: MW = 36.47
EW = 36.47
Specific gravity = 1.18
Commercially available concentrated hydrochloric acid has strength of 36 to 39 percent and is

10.5 to 12 N.
1. Measure by means of graduated cylinder 9 ml of analytical reagent concentrated HCl.

2. Pour into 1-liter volumetric flask.
3. Dilute to 1 liter with distilled water and mix well.
Standardization with 0.1N Sodium Carbonate
1. Dry pure analytical reagent sodium carbonate (anhydrous) Na2CO3 in a crucible in an oven
at 280oC for 30 minutes. Cover the cylinder and place it in a desiccator.
2. Weigh exactly 1.3249g dried sodium carbonate in analytical balance. Dissolve in water,
dilute to 250ml in 250 ml volumetric flask and mix well. The equivalent weight of sodium
carbonate is 52.997. Therefore, to prepare 0.1 N sodium carbonate in a liter, 5.2997g will
have to be dissolved in 1-liter water.
3. Rinse a clean burette with the acid to be titrated. Fill the burette with the acid.
4. Pipette 25 ml of the sodium carbonate solution in to a 100ml Erlenmeyer flask.
5. Add 3 drops methyl orange indicator.
6. Titrate with the acid until the color of the solution becomes faint yellow. Continue addition
of acid drop by drop till the color changes to orange or faint pink, note the burette reading.
Repeat the titration 2 more times. The difference in reading between titration should not be
more than 0.1 ml. Calculate the normality of the acid using the formula:

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𝑁1𝑉1 = 𝑁2𝑉2
Where N1 is the normality of sodium carbonate,

V1 is the ml of sodium carbonate and
V2 is the ml of acid required for titration.
Then, N2 = N1V1 = 25x0.1

V2 V2
Preparation of 0.1N Standard Sodium Hydroxide
Generally, sodium hydroxide will contain small amounts of sodium carbonate. Even the
analytical reagent quality (AR) may contain about 1 to 2 percent sodium carbonate. If it is
necessary to obtain carbonate free sodium hydroxide, the carbonate will have to be precipitated
with barium chloride. Otherwise, make a concentrated solution of sodium hydroxide and leave
it for some time. The carbonate will form a crust on the surface of the solution. However, the
AR quality chemical can be used without removing the carbonates first for all practical purposes.
1. Weigh 4.2 g AR sodium hydroxide (NaOH), M.W = 40, E. W= 40) in a beaker, dissolve in
water and dilute to 1 litre with freshly boiled and cooled (to remove carbon dioxide) distilled
water.
2. Mix thoroughly by shaking and pour the solution in to a bottle and close the bottle tight with
a rubber stopper.
Standardization of Sodium Hydroxide with Potassium Hydrogen Phthalate
Potassium hydrogen phthalate is a primary standard. It has a formula KH (C2H4O4) and

molecular weight of 204.22 and equivalent weight of the same (204.22).
1. Dry some Potassium hydrogen phthalate in an oven and cool in a desiccator.
2. Weigh exactly 5.1055 g of Potassium hydrogen phthalate, dissolve in water and dilute to 250
ml in a volumetric flask. Mix well.
3. Pipette 25 ml potassium hydrogen phthalate solution in to a 100 ml Erlenmeyer flask.

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4. Add 2 drops phenolphthalein indicator.

5. Titrate with sodium hydroxide solution till the color turns pink. Note the burette reading.
Repeat 3 times and take the average.
KH(C8H4O4) + NaOH KNa(C8H4O4) + H2O
Calculate the normality of NaOH as N = (25 x 0.1) / ml NaOH required for titration.
Preparation of 0.1 N Silver Nitrate Solutions
AgNO3: MW = 169.87
EW = 169.87
1. Dry some finely powdered silver nitrate in an oven at 120oC for 2 hours and cool in a
desiccator.
2. Weigh 8.496 g and dissolve in water.
3. Make up to 500 ml in a volumetric flask. Store in a brown colored bottle (amber glass).
Standardization of Silver Nitrate with Sodium Chloride
NaCl: M.W = 58.46

E.W = 58.46
1. Prepare 0.1 N NaCl by dissolving 2.923 g pure, A.B. sodium chloride in water and diluting
to 500 ml.
2. Dissolve 5 g potassium chromate (K2CrO4) in 200 ml water. Add a few ml silver nitrate
solutions. A red precipitate will appear. Keep for a day in a dark place; filter.
3. Pipette 25 ml 0.1 N sodium chloride solution in to a 100 ml Erlenmeyer flask. Add 1 ml
potassium chromate solution.
4. Titrate with silver nitrate until a faint red color persists. Note the reading. Calculate the
normality of silver nitrate in the same way as the above standards.

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Preparation of 0.1N Potassium Permanganate
KMnO4: M.W = 158.04

No of Eqv. = 5
E.W = 31.608
1. Weigh 3.2 g A.R quality potassium permanganate. Transfer to 1 litre beaker. and make to
the mark with distilled water
2. Stir till the entire solid dissolves. Cover the beaker with a watch glass.
3. Heat to boiling on a hot plate. Boil gently for about 30 minutes. Cool. Filter. Store in a dark
colored bottle.
4. Standardize with sodium oxalate.
Preparation of 0.5N Ferrous Ammonium Sulfate
Fe (NH4) 2(SO4)2.6H2O: M.W = 392.13

E.W = 392.13
1. Weigh 196.065g pure AR ferrous ammonium sulfate and dissolve in water.

2. Add 20 ml con. Sulfuric acid and cool.
3. Dilute to 1 litre with water.
Preparation of 1N Potassium Dichromate
K2Cr2O7; M.W = 294.22

No of Eqv = 6
E.W = 49.03
1. Grind about 100 g potassium dichromate (A.R. grade) in a glass mortar.

2. Dry for 30 minutes at 150oC in an oven. Cool in a desiccator.
3. Weigh 49.03 g and transfer to a 1 litre volumetric flask. Dissolve in water and dilute to 1
litre. Mix well.

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Standardize with Potassium Dichromate
1. Pipette 10 ml 1 N potassium dichromate in to a 500 ml Erlenmeyer flask.

2. Add 100 ml distilled water and 10 ml phosphoric acid.
3. Add 1 ml diphenylamine indicator. The color of the solution will turn deep blue.
4. Titrate slowly with ferrous ammonium sulfate till the color changes to emerald green (bright
green or also called pea green).
5. Note the burette reading and calculate the normality of ferrous ammonium sulfate.
Preparation of 0.1N EDTA
Disodium ethylene diamine tetra acetic acid:

(Na2H2C10H12O8N2. 2H2O): M.W = 372.25
E.W = 186.125
1. Dissolve 18.6125 g pure EDTA in water.
2. Dilute to 1 litre in a volumetric flask. Shake well.
3. Standardize with calcium standard
Preparation of 0.1N Calcium Carbonate Solution from Calcium Carbonate
CaCO3; M.W = 100.09

E.W = 50.04
1. Weigh 5.005g pure dry calcium carbonate powder and place in a beaker.
2. Add enough 1N (1:12) hydrochloric acid drop by drop to dissolve the carbonates.
3. Wash down the side of the beaker with a stream of water.
4. Transfer to a 1 litre volumetric flask. Make up to 1 litre with water and mix well.
Preparation of 0.1N Sulfuric Acid (H2SO4)
H2SO4: MW = 98
EW = 49
Specific gravity = 1.84

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Normality = 36
1. Take about 500 ml distilled water in a beaker. Add 2.8 ml concentrated H2SO4 to it slowly.
Pour the acid through the sides of the beaker.
2. Mix by stirring and let the solution become cool.
3. Transfer to a 1 litre volumetric flask and dilute to 1 litre with water
4. Standardize as for HCl.
Calculation of the Amount of an Acid Needed to Prepare Standard Solution
E.g. Prepare 1000 ml 0.5 N sulfuric acid

Sulfuric acid, formula = H2SO4.
Molecular weight (M.W.) = 2+32+64 =98.
Equivalent weight (E.W) = 98/2 = 49.
% Acid in the commercial acid = 96 (read from the label on bottle).
Specific gravity = 1.84.
To prepare 1 liter of 1 N sulfuric acid, 49 g acid is needed, provided the acid is 100 percent
strong. But the acid is only 96% strong. Therefore, the weight of acid needed to prepare 1 litre
1N solution would be 49x100/96
 Weight of acid for 1 litre 0.5N acid will be:

49 x 100 x 0.5 = 25.52 g
96
 Volume of acid needed = weight /specific gravity:
= 49 x 100 x 0.5 x 1 ml = 25.52 = 13.87 ml

96 1.84 1.84
In general, the following formula can be used:
𝐸𝑞. 𝑊𝑡. 𝑥 100 𝑥 𝑁 𝑥 𝑉

𝑚𝑙 𝑜𝑓 𝑎𝑐𝑖𝑑 𝑛𝑒𝑒𝑑𝑒𝑑 (𝑚𝑙) =
% 𝐴𝑐𝑖𝑑 𝑥 𝑆𝑝. 𝐺𝑟 𝑜𝑓 𝐴𝑐𝑖𝑑

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Where: E.W is the equivalent weight of the acid.

N is the normality needed.
V is the volume in litre of the solution needed.
Sp.gr is the specific gravity of the concentrated acid to be used.
5. Dilution of Solutions and Dilution Methods
Because solutions in science are often much more concentrated than are desired or can be
managed for a given protocol, it is frequently necessary to dilute these solutions to a desired
level. This requires a working knowledge of the principles of diluting, dilution factors,
concentration factors and the calculations involved. High dilutions are usually expressed
exponentially (i.e. a solution which has been diluted a million-fold is termed a 106 dilution, or is
10-6 concentration.)
Definitions
✓ Aliquot: a measured sub-volume of original sample.

✓ Diluent: material with which the sample is diluted.
✓ Dilution factor (DF): ratio of final volume/aliquot volume (Final volume = Aliquot +
Diluent).
✓ Concentration factor (CF): ratio of aliquot volume divided by the final volume (inverse
of the dilution factor).
How to Calculate Dilution Solutions
A dilution solution contains solute (or stock solution, aliquot) and a solvent (called diluent).
These two components proportionally combine to create a dilution. You can identify a dilution
solution by the amount of solute in the total volume, expressed as a proportion. For example, a
chemical may be prepared in a 1:10 dilution of alcohol, indicating that a 10 mL bottle contains
one milliliter of chemical and nine milliliters of alcohol. We can calculate the necessary volume
of each component to prepare a dilution solution and calculate the dilution factor based on known
volumes of the solution.

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Instructions
Things you’ll need:
✓ Graduated cylinder.
✓ Measuring cup (large and small).
✓ Calculator.
Prepare Dilution Solution
Step 1
✓ Write down the final volume of the solution--for example, 30 mL.
Step 2
✓ Write down the desired dilution in the form of a proportion--for example, 1:20 dilution,
also known as the dilution factor.
Step 3
✓ Convert the dilution factor to a fraction with the first number as the numerator and the
second number as the denominator. For example, a 1:20 dilution converts to a 1/20
dilution factor.
Step 4
✓ Write down the formula to determine the needed volume of the stock solution, also known
as the aliquot: final volume (in step 1) x dilution factor (step 3) -- for example, 30 mL x
1/20 from steps 1 and 3, respectively.
Step 5
✓ Calculate the amount of stock solution required--for example, 30 mL x 1/20 = 1.5 mL.
Step 6
✓ Subtract the amount of stock solution (step 5) from the total volume (step 1) to calculate
the volume of diluent required--for example, 30 mL - 1.5 mL = 28.5 mL.
Step 7
✓ Measure the amount of stock solution required (step 5) and dispense this into a large
measuring cup. For example, dispense 1.5 mL of aliquot into a 50-mL measuring cup.

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Step 8
✓ Measure the amount of diluent required (step 6) and dispense this into the large measuring
cup. For example, dispense 28.5 mL of diluent into the 50-mL measuring cup containing
1.5 mL aliquot to obtain the desired dilution solution.
Calculate Dilution
Step 1
✓ Measure the total volume of the solution--for example, 25 mL.
Step 2
✓ Write down the volume of stock solution (or aliquot) or diluent used in the solution--for
example, 10 mL of aliquot; alternatively, 20 mL diluent.
Step 3
✓ Subtract the diluent volume (step 2) from the total volume of the solution (step 1) to
calculate the volume of aliquot used, if the volume of stock solution (aliquot) isn't known-
-for example, 25 mL total volume - 20 mL diluent = 5 mL aliquot.
Step 4
✓ Write down the proportion for the dilution--dilution factor = volume of aliquot: total
volume.
Step 5
✓ Insert the known values to calculate the dilution factor and simplify. For example:
Aliquot from step 2 = 1 mL. Dilution factor = 10 mL:25 mL simplifies to 2:5.
Aliquot from step 3 = 5 mL. Dilution factor = 5 mL: 25 mL = simplifies to 1:5.
How to Make Simple Dilutions
Simple Dilution (Dilution Factor Method Based on Ratios)
A simple dilution is one in which a unit volume of a liquid material of interest is combined with
an appropriate volume of a solvent liquid to achieve the desired concentration. The dilution
factor is the total number of unit volumes in which your material will be dissolved. The diluted
material must then be thoroughly mixed to achieve the true dilution. For example, a 1:5 dilution

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(verbalize as "1 to 5" dilution) entails combining 1unit volume of solute (the material to be
diluted) + 4unit volumes of the solvent medium (hence, 1 + 4 = 5 = dilution factor). The dilution
factor is frequently expressed using exponents: 1:5 would be 5e-1 (5-1); 1:100 would be 10e-2
(10-2), and so on.
Example 1: Frozen orange juice concentrate is usually diluted with 4 additional cans of cold
water (the dilution solvent) giving a dilution factor of 5, i.e., the orange concentrate represents
one-unit volume to which you have added 4 more cans (same unit volumes) of water. So, the
orange concentrate is now distributed through 5-unit volumes. This would be called a 1:5
dilution, and the Orange Juice is now 1/5 as concentrated as it was originally. So, in a simple
dilution, add one less unit volume of solvent than the desired dilution factor value.
Example 2: Suppose you must prepare 400 ml of a disinfectant that requires 1:8 dilutions from
a concentrated stock solution with water. Divide the volume needed by the dilution factor (400
ml / 8 = 50 ml) to determine the unit volume. The dilution is then done as 50 ml concentrated
disinfectant + 350 ml water.
Serial Dilution
A serial dilution is simply a series of simple dilutions which amplifies the dilution factor quickly
beginning with a small initial quantity of material (i.e., bacterial culture, a chemical, orange juice,
etc.). The source of dilution material for each step comes from the diluted material of the
previous. In a serial dilution the total dilution factor at any point is the product of the individual
dilution factors in each step up to it.
𝐹𝑖𝑛𝑎𝑙 𝐷𝑖𝑙𝑢𝑡𝑖𝑜𝑛 𝐹𝑎𝑐𝑡𝑜𝑟 (𝐷𝐹) = 𝐷𝐹1 × 𝐷𝐹2 × 𝐷𝐹3 . ..
Example: In a typical microbiology exercise the students perform a three step 1:100 serial
dilution of a bacterial culture (see figure below) in the process of quantifying the number of
viable bacteria in a culture (see figure below). Each step in this example uses a 1 ml total volume.
The initial step combines 1-unit volume of bacterial culture (10 l) with 99-unit volumes of broth
(990 l) = 1:100 dilution. In the second step, 1-unit volume of the 1:100 dilution is combined
with 99-unit volumes of broth now yielding a total dilution of 1:100x100 = 1:10,000 dilution.

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Repeated again (the third step) the total dilution would be 1:100x10, 000 = 1:1,000,000 total
dilution. The concentration of bacteria is now one million times less than in the original sample.
Making fixed volumes of specific concentrations from liquid reagents
𝑉1𝐶1 = 𝑉2𝐶2
Very often you will need to make a specific volume of known concentration from stock solutions,
or perhaps due to limited availability of liquid materials (some chemicals are very expensive and
are only sold and used in small quantities, e.g., micrograms), or to limit the amount of chemical
waste. The formula below is a quick approach to calculating such dilutions, where V = volume,
C = concentration; in whatever units you are working.
(𝑆𝑡𝑜𝑐𝑘 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 𝑎𝑡𝑡𝑟𝑖𝑏𝑢𝑡𝑒𝑠) 𝑽𝟏𝑪𝟏 = 𝑽𝟐𝑽𝟐 (𝑁𝑒𝑤 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 𝑎𝑡𝑡𝑟𝑖𝑏𝑢𝑡𝑒𝑠)
Example: Suppose you have 3 ml of a stock solution of 100 mg/ml ampicillin (= C1) and you
want to make 200 l (= V2) of solution having 25 mg/ ml (= C2). You need to know what volume
(V1) of the stock to use as part of the 200 l total volume needed.
V1 = the volume of stock you’ll start with. This is our unknown.
C1 = 100 mg/ ml in the stock solution

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V2 = total volume needed at the new concentration = 200 ul = 0.2 ml

C2 = the new concentration = 25 mg/ ml
By algebraic rearrangement:
V1 = (V2 x C2) / C1
V1 = (0.2 ml x 25 mg/ml) / 100 mg/ml and after canceling the units,
V1 = 0.05 ml or 50 l
So, you would take 0.05 ml = 50 l of stock solution and dilute it with 150 l of solvent to get
the 200 l of 25 mg/ml solution needed (remember that the amount of solvent used is based upon
the final volume needed, so you have to subtract the starting volume from the final to calculate
it.)
6. Volumetric Analysis
It is a general term for a method in quantitative chemical analysis in which the amount of a
substance is determined by the measurement of the volume that the substance occupies. It is
commonly used to determine the unknown concentration of a known reactant.
Volumetric analysis is often referred to as titration, a laboratory technique in which one
substance of know concentration and volume is used to react with another substance of unknown
concentration. For use in titrimetric analysis a reaction must satisfy the following conditions:
✓ There must be a simple reaction that can be expressed by a chemical equation; the
substance to be determined must react completely with the reagent in the stoichiometric
or equivalent proportions.
✓ The reaction kinetics must be rapid. In certain cases, addition of a catalyst increases the
speed of reaction.
✓ There must be a marked change in free energy leading to alteration in some physical or
chemical properties of the solution at the end point.
✓ An indicator must be available which should sharply define the end point of reaction by
a change in color or formation of precipitate etc.

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Definition of Terms
Titration: A process in which a standard reagent is added to a solution of analyte until the
reaction between the two is judged complete. The reagent of known concentration is known as
titrant and the substance being titrated is known as titrand.
Standardization: A process to determine the concentration of a solution of known concentration
by titrating with a primary standard
End Point
✓ The point at which the reaction is observed to be completed is the end point
✓ The end point in volumetric method of analysis is the signal that tells the analyst to stop
adding reagent and make the final reading on the burette.
✓ Endpoint is observed with the help of indicator
Equivalent Point
✓ The point at which an equivalent or stoichiometric amount of titrant is added to the
analyte based on the stoichiometric equation.
Types of Volumetric Analysis
The reactions employed in titrimetric analysis may be broadly divided into two main classes:
✓ Those in which no change in oxidation states occurs; these are dependent upon
combination of ions.
✓ Oxidation – reduction (redox) reactions involving electron transfer or change of
oxidation states.
For simplicity and convenience, the above two broad categories are subdivided into four main
classes:

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Neutralization Reactions or Acidimetry and Alkalimetry
The two terms, (acidimetry and alkalimetry) are complementary. They involve determination
of concentration of acid or an alkali solution by titration against a standard solution of alkali or
an acid as case may be. If the concentrations in normality of the alkali and the acid solutions are
NA and NB respectively and VA ml of the alkali exactly neutralizes VB ml of the acid. Then,
𝑉𝑎 . 𝑁𝑎 = 𝑉𝑏 . 𝑉𝑏
This is the fundamental equation of acidimetry-alkalimetry. If the strength of one is known, the
other can be calculated out.
6.2.2 Theory of Acid Base Titration
Neutralization indicators are substances which exhibit different colors according to the H+ ion
concentration or pH of the solution to which they are added. It is thus possible to have an idea
of the pH of a given solution by adding a little of suitable indicator to the same. Moreover, if an
indicator is present during the progress of a titration of an alkali with an acid, the color change
of the indicator reveals the end point of titration. Most of the indicators have a predominantly
‘acid color’ and a predominantly ‘alkaline color’ in the lower and higher ranges of pH. The chief
characteristics of such indicators is that the change from a predominantly ‘acid color’ to a
predominantly ‘alkaline color’ is not sudden and abrupt, but takes place within a small interval
of pH, termed as color change interval of the indicators. For most acid-base titration we can
therefore select an indicator which exhibits a distinct color change at pH close to that obtaining
at the equivalence point.

The terms “acidimetry” and “alkalimetry” are derived from the words acid and alkali. In determination of an acid
the solution is titrated with an alkali solution the volume of which is measured by means of a burette. This accounts
for the name alkalimetry. The name of the method for determining the amount of alkali, acidimetry, is derived
analogously. However, this terminology is not universally accepted; some writers (for example, Treadwell) call
determination of acids acidimetry, and determination of alkalies, alkalimetry.

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Indicators are either very weak organic acids or bases which always exist in two tautomeric
forms. One of the tautomers is in the non-electrolytic forms and scarcely ionizes but the other is
an electrolyte and hence ionizable.
Let HIn’ be the non-ionizable tautomeric form and HIn be the ionizable form of phenolphthalein
where In- represents the indicator ion. The color of HIn and In- are the same but different from
HIn’. In aqueous solution there would exist two equilibrium.
HIn’ HIn; 𝐾𝑡 = 𝑎𝐻𝐼𝑛 ⁄𝑎𝐻𝐼𝑛′
HIn H+ + In-; 𝐾𝐷 = 𝑎𝐻 + 𝑎𝐼𝑛+ ⁄𝑎𝐻𝐼𝑛
In acid solutions the dissociation will be suppressed and the whole of the indicator ion shall
remain in undissociated form i.e. as HIn’ and HIn, which have different colors. The indicators
will be applicable only if the equilibrium constant Kt is small that the undissociated indicator
mostly exists as HIn’. Multiplying the above two equilibrium constants by each other gives:
𝑎𝐻 + . 𝑎𝐼𝑛−
𝐾𝑡 . 𝐾𝐷 = = 𝐾𝑖𝑛
𝑎𝐻𝐼𝑛
Where Kin is called the indicator constant, it has been assumed that the concentration (hence
activity) of HIn is very small. Therefore, aHIn is the activity of practically the entire undissociated
form. The color of HIn’ is the color which the indicator will show in acid medium. In presence
of alkali, H+ ions will be removed as H2O, dissociation will be almost complete and indicator
will present mostly as In-. Hence indicator ion shall have ‘alkali’ color. Rewriting the above
equation,
𝑎
𝑎𝐻 + = 𝐾𝑖𝑛 . 𝑎𝐻𝐼𝑛−
𝐼𝑛
𝐶 𝛾
𝑎𝐻 + = 𝐾𝑖𝑛 𝐶𝐻𝐼𝑛− . 𝛾𝐻𝐼𝑛−
𝐼𝑛 𝐼𝑛

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Or
𝐶𝐼𝑛−
𝑝𝐻 = 𝑝𝐾𝑖𝑛 + 𝑙𝑜𝑔 + 𝑙𝑜𝑔 𝛾𝐼𝑛− ; [𝛾𝐻𝐼𝑛 = 1]
𝐶𝐻𝐼𝑛
Log In- may be evaluated with extended Debye-Huckel law, but for most purposes, it is small
and can be neglected, so that:
𝐶 − 𝐶𝑖𝑜𝑛𝑖𝑧𝑒𝑑 𝑓𝑜𝑟𝑚 𝑤𝑖𝑡ℎ 𝑎𝑙𝑘𝑎𝑙𝑖𝑛𝑒 𝑐𝑜𝑙𝑜𝑟

𝑝𝐻 = 𝑝𝐾𝑖𝑛 + 𝑙𝑜𝑔 𝐶 𝐼𝑛 Or 𝑝𝐻 = 𝑝𝐾𝑖𝑛 + 𝑙𝑜𝑔 𝐶
𝐻𝐼𝑁 𝑛𝑜𝑛−𝑖𝑜𝑛𝑖𝑧𝑒𝑑 𝑓𝑜𝑟𝑚 𝑤𝑖𝑡ℎ 𝑎𝑐𝑖𝑑 𝑐𝑜𝑙𝑜𝑟
It is seen that at a given pH, indicator will exist in a definite ratio of concentrations of ionized
and non-ionized form. Both the forms are present in any pH, but human eye can discern the color
distinctly when one predominates. It has been found that the acid color, namely that of HIn’, is
𝐶𝐻𝐼𝑛
detected when > 10, i.e. when pH = pKin – 1 and the alkaline color can be detected when
𝐶𝐼𝑛−
𝐶𝐼𝑛−
> 10, i.e. pH = pKin + 1.
𝐶𝐻𝐼𝑛
As we titrate acid with base, the pH changes. The indicator will change from one color to another
within the pH range (pKin + 1) to (pKin – 1) within two units of pH. Thus, phenolphthalein has a
pKin value of 8.96 and hence its color change takes place in the pH range 7.96 to 9.96. Methyl
red has a pHin value of 5.1, so its color would change in the pH range 4.1 to 6.1.
Principle of Acidimetry and Alkalimetry
Acidimetry is the method of determining the concentration of an acid by titrating with a standard
solution (i.e. known concentration) of an alkali. The method with help of which the concentration
of an alkali solution is found out by titrating with a standard solution of an acid is alkalimetry.
The law of normality can be stated as: “Equal volumes of solutions of two reacting substances
(acids and bases) of the same concentration expressed in normality exactly neutralize each other”
i.e., V volume of X normal solution of any acid will exactly neutralize V volume of X normal
solution of any base. The concentration of any solution decreases with dilution. In fact, the
concentration and volume of a solution are inversely proportional.

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1
Therefore, volume (V) of a solution  𝑐𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 (𝑠) 𝑜𝑓 𝑡ℎ𝑒 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
If V1 volumes of an acid of exact concentration C1 neutralizes V2 volumes of a base of

𝑉 𝐶
concentration C2, then, 𝑉1 = 𝐶2 or V1C1 = V2C2.
2 1
Complex Formation Reaction
These depend upon the combination of ions other than hydrogen or hydroxide ions, to form a
soluble slightly dissociated ion or compound. Ethylene diamine tetra acetic acid, mostly as
disodium salt of EDTA, is a very important reagent for complex formation. The use of metal
ion-indicators has highly enhanced its importance in titrimetry.
Precipitation Reaction
Such reactions depend upon the combination of ions resulting in formation of a simple precipitate
as in the titration of silver ion with a solution of a chloride. No change in oxidation state occurs.
Oxidation-Reduction Reaction
All reactions involving change in oxidation number or transfer of electrons among the reacting
substances fall under this category. The standard solutions are either oxidizing or reducing
agents. The principal oxidizing agents are potassium dichromate, potassium permanganate,
potassium iodate, etc., while common reducing agents are sodium thiosulfate, Iron (II) and tin
(II) compounds, chromium (II) chloride or sulfate etc.
7. Indicators
A substance which indicates the ‘end point’ or completion of reaction is known as indicator. It
is an auxiliary reagent which helps in the visual determination of the completion of titration.
Generally, three types of indicators are used in volumetric analysis.

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Internal Indicator
The indicators are added into the solution where the reaction occurs, which gives a clear visual
change in the solution being titrated, e.g. methyl orange, methyl red, phenolphthalein, etc.
The internal indicators are also divided into the following groups according to their use in
different types of reactions.
Indicators used in Acid-Alkali Neutralization Reaction:
Some examples of such indicators are methyl red, phenolphthalein, methyl orange, bromothymol
blue, etc.
These are normally known as hydrogen ion or acid-base indicators. Actually, such indicators are
organic dyes (weak organic acids or bases) which change s color s within limits with variation
in pH-value of the solution to which it is added. The important characteristics of these indicators
are that the change from a predominantly ‘acid’ color to a predominantly ‘alkaline’ color is not
sudden and abrupt, but takes place within a small interval of pH termed as the color change
interval of the indicator. It is advisable, to select the indicator which exhibits a distinct color
change at a pH close to that obtaining at the equivalence for acid-base titration.
Indicators used in Precipitation Reaction:
In the titration of sodium chloride with silver nitrate, a small quantity of K 2CrO4 solution is
added to serve as indicator. At the end point the chromate ion combines with Ag+ ions to form
sparingly soluble Ag2CrO4 (brick red color). Both AgCl and Ag2CrO4 are insoluble, but they are
precipitated only when their respective solubility products are reached (AgCl: 1.2 x 10-10,
Ag2CrO4: 1.7 x 10-12). As long as chloride ions are present in sufficient concentrations in the
titration mixture, only AgCl is precipitated and no Ag2CrO4 precipitates. When all Cl- ions are
removed as AgCl only then a red precipitate of Ag2CrO4 appears and gives a reddish or brownish
end point.

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Redox Indicators:
Redox indicator is a substance which possesses a different color in the oxidized form and a
different color in the reduced form. Some organic dye stuff belongs to this class. In order to get
a sharp color change at the end point the indicator chosen for a particular titration must have its
standard potential (Eo) values in between the standard potential of the oxidation potential of the
oxidation-reduction systems being titrated against each other. Examples are diphenylamine,
methylene blue, diphenylamineazo sulfonic acid, etc. Redox indicator is a substance which
possesses a different color in the oxidized form and a different color in the reduced form. Some
organic dye stuffs belong to this class. In order to get a sharp color change at the end point the
indicator chosen for a particular titration must have its standard potential (Eo) values in between
the standard potential of the oxidation-reduction systems being titrated against each other.
Examples are diphenylamine, methylene blue, diphenylamineazo sulfonic acid, etc.
In a redox titration of a reductant with an oxidant there occurs a continuous change of potential
of a solution due to a gradual change of concentration of the species involved in the reaction. It
can be easily shown that the potential changes abruptly in the neighborhood of equivalence point
and is dependent upon the standard potentials of the two oxidation-reduction systems involved
but independent of the concentrations. In general for the reaction a, 𝑎𝑂𝑥𝐼 + 𝑏𝑅𝑒 𝑑𝐼𝐼 → 𝑏𝑂𝑥𝐼𝐼 +
𝑎𝑅𝑒 𝑑𝐼 the oxidation potential at the equivalence point is given by:
(𝑏𝐸𝐼 𝑜 +𝑎𝐸𝐼𝐼 𝑜 )
𝐸𝑜 = ( ), where, EIo and EIIo are the standard oxidation potentials
𝑎+𝑏
of the systems RedI/ OxI and RedII/ OxII.

An oxidation-reduction indicator is a substance which can mark the sudden change in the
oxidation potential in the neighborhood of the equivalence point in a redox titration. The ideal
redox indicator will be one with an oxidation potential intermediate between that of the solution
titrated and that of the titrand. A redox indicator is a compound which exhibits different colors
in the reduced and oxidized forms.
𝐼𝑛𝑟𝑒𝑑 → 𝐼𝑛𝑜𝑥
At a potential E, the ratio of concentrations of the oxidized and reduced forms is given by:

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𝑅𝑇 [𝐼𝑛 ]
𝐸 = 𝐸𝐼𝑛 𝑜 − 𝑛𝐹 𝑙𝑛 [𝐼𝑛 𝑜𝑥 ]
𝑟𝑒𝑑
Where EIno is the standard oxidation potential if the color intensities of the two forms are
comparable a practical estimate of the color change interval corresponds to change of the
[𝐼𝑛 ]
ratio[𝐼𝑛 𝑜𝑥 ]from 10 to 1/10. The interval of potential is thus;
𝑟𝑒𝑑
𝐸 𝑜 𝐼𝑛 ±0.059
𝐸=( ) 𝑣𝑜𝑙𝑡 𝑎𝑡 25𝑜 𝐶
𝑛
For a color change at the end point, EoIn should differ by about at least 0.15 V from the standard
(formal) potentials of the other systems involved in the reaction.
The earliest known redox indicator is diphenylamine used for titration of Fe (II) with potassium
dichromate. An intense blue violet coloration is produced at the end point. The action of
diphenylamine (I) depends upon its oxidation first into colorless diphenyl benzidine (II) which
is the real indicator and reversibly further oxidized to diphenyl benzidine (III), (violet). Formal
oxidation potential of (II)/ (III) system is E’oIn = - 0.76 volt and n = 2.
Hence the potential range of color change of the indicator is (-0.76  0.059/2) = -0.73 volt to -
0.79 volt. Above -0.73 volt the reduced form is predominant and the solution is colorless while
at -0.79 volt or below the oxidized form predominates and the solution assumes blue-violet color.
Now standard oxidation potential (Eo) of Fe2+/Fe3+ and 2Cr3+/Cr2O72- systems are -0.77 and -
1.33, and the sharp fall of potential near the equivalence point occurs in the range -0.944 volt to
-1.302 volt. Evidently diphenylamine is not a suitable indicator, However addition of phosphoric
acid complexes Fe (III) as Fe (HPO4) + ion. This increases the formal potential of Fe (II)/Fe
(III) system so that the equivalence point potential coincides more nearly with that of the
indicator. The limits of sharp change of potential for color change of diphenylamine sulfonate is
to some extent preferable, firstly because it is water soluble and secondly, it’s formal oxidation
potential (-0.85 V in 0.5 M H2SO4) is somewhat lower than that of diphenylamine.

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External Indicator
These are not added to the reacting medium. For example, potassium ferricyanide, K3Fe (CN)
6 is used as an indicator in the titration of potassium dichromate with ferrous sulfate in an acid
medium. K3Fe (CN) 6 reacts with Fe2+ ion and forms a deep blue color compound of ferro-
ferricyanide.
2𝐾3 [𝐹𝑒(𝐶𝑁)6 ] + 3𝐹𝑒 ++ → 𝐹𝑒3 [𝐹𝑒(𝐶𝑁)6 ]2

𝑑𝑒𝑒𝑝𝑏𝑙𝑢𝑒
The indicator gives blue color with Fe++ ions, hence when all the ferrous ions are oxidized in
solution, it will not give the blue color.
Self-Indicator
When one of the reactants itself acts as an indicator by visual color change, is called a self-
indicator or auto-indicator. KMnO4 acts as an auto-indicator whenever titrated with oxalic acid
or ferrous sulfate in presence of dilute H2SO4.
Furthermore, there are adsorption and complex forming indicators. In iodometric titration starch
solution is used as an indicator which forms a complex with I2, which has a very dark blue color.
Choice of Indicators
Strong Acid against a Strong Base
For example, titration of HCl and NaOH. At the initial stage of titration, the pH changes very
slowly and increases up to pH= 4. But with addition of small amount of alkali like 0.01 mL
approx., pH value reaches to about 7. This shows the neutralization of acid. Further addition of
small amount of alkali increases the concentration of hydrogen ions and the pH value rises to
about 9. There is a rapid increase of pH from about 4 to 9 when the end point approaches.
Indicators like methyl orange, methyl red and phenolphthalein could be used for these kinds of
neutralization reactions which show the color change within the pH range of 4 to 10.

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Weak Acid against Strong Base
Example: Titration of acetic acid against NaOH. The end point of this titration lies between pH
8 and 10 due to the formation of sodium acetate and water. So, phenolphthalein is a good
indicator for this.
Strong Acid against Weak Base
For example: the titration of ammonium hydroxide with HCl. The end point is in the range of 6
to 4 which shows the acidic pH of solution. Methyl orange is a good indicator for this titration.
Table 2: Acid Base Indicators
Indicator Color change on pH Range of color Color change on basic

acidic side change side
Methyl Violet Yellow 0.0 - 1.6 Violet
Bromophenol Blue Yellow 3.0 - 4.6 Blue
Methyl Orange Red 3.1 - 4.4 Yellow
Methyl Red Red 4.4 - 6.3 Yellow
Litmus Red 5.0 - 8.0 Blue
Bromothymol Blue Yellow 6.0 - 7.6 Blue
Phenolphthalein Colorless 8.3 - 10.0 Pink
Alizarin Yellow Yellow 10.1 - 12.0 Red

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8. References
1. Chem.74(5), pp 835855, http://www.iupac.org/publications/pac/2002/7405/x0835.html

2. Douglas, S. A., Donald, W. M., F, H. J., & Stanley, R. C. (2004). Fundamentals of Analytical
Chemistry (Eighth ed.). Thomson Books.
3. John, K. (2002). Analytical chmistry for technicians . CRC press LLC.
4. Eurachem Guide (1998), The fitness for purpose of analytical Methods. A laboratory guide
to method validation and related topics, Edition 1.0, http://eurachem.ul.pt/guides/mval.html
5. F, F. W., & D, K. (2000). Principle and practice of analytical chemistry.
6. David, H. (2000). Modern analytical chemistry. Depauw university: MC Graw Hill.
7. IUPAC (2002), Harmonized guidelines for single laboratory validations of methods of
analysis, Pure Appl.
8. JCGM: 200 (2008), International Vocabulary of Metrology (VIM) – Basic and general
concepts and associated terms, 3rd edition, BIPM, Sèvres, www.bipm.org/vim2008

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LIST OF TABLES ................................................................................................................................................II

1. Introduction to Analytical Chemistry

“Analytical chemistry is what analytical chemists do.”

1.1. The Nature of Analytical Chemistry

Analytical chemistry is a measurement science consisting of a set of powerful ideas and

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Figure 1 Flow diagram showing the steps in a quantitative analysis

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e) Calibration and measurement

1.2. The Role of Analytical Chemistry

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Figure 2 The relationship between analytical chemistry, other branches of chemistry

1.3. Classification of Chemical Analysis

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Classical Vs. Instrumental Techniques

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Single-Channel Vs. Multi-Channel Techniques

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1.4. Methods of Analysis

1.4.1. Gravimetric Methods of Analysis

Types of Gravimetric Methods

• Precipitation gravimetry: A gravimetric method in which the signal is the mass of a

1.4.2. Titrimetric Methods of Analysis

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Types of Titrimetric Methods

1.4.3. Spectroscopic Methods of Analysis

Spectroscopy is a branch of science that studies the interaction between electromagnetic

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Spectroscopy Based on Absorption

In absorption spectroscopy a beam of electromagnetic radiation passes through a sample. Much

Spectroscopy Based on Emission

Spectroscopy Based on Scattering

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1.4.4. Electrochemical Methods of Analysis

Analytical methods in which a measurement of potential, current, or charge in an

1.4.5. Chromatographic and Electrophoretic Methods of Analysis

Chromatography is a separation in which solutes partition between a mobile and stationary

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1.5. Basic Terminologies in Analytical Chemistry

𝐿𝑂𝐷 = 𝑀𝑒𝑎𝑛 + 3𝑆𝐷

Limit of Quantitation, Limit of Determination: Refers to the smallest analyte concentration or

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𝐿𝑂𝑄 = 𝑀𝑒𝑎𝑛 + 10𝑆𝐷

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measurements and can be characterized by experimental standard deviations. Estimates of other

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1.6. Error in Chemical Analysis

1.6.1. Types of Error

i. Random (Indeterminate) Error

1.6.1.1. Random (Indeterminate) Errors

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1.6.1.2. Systematic (Determinate) Errors

Sources of Systematic Error

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1.6.1.3. Gross Errors

2. Concentration and Methods of Expressing Concentration

2.1. Molarity (M)