STUDY MATERIAL

BSc /BSc (Honours) – I BSc DSC

Analytical Chemistry
Unit –I 14 Hrs

Basic laboratory practices, calibration of glassware (pipette, burette and volumetric flask),
Sampling (solids and liquids), weighing, drying, dissolving, Acid treatment, Rules of work in
analytical laboratory, General rule for performing quantitative determinations (volumetric and
gravimetric), Safety in Chemical laboratory, Rules of fire prevention and accidents, First aid.
Precautions to be taken while handling toxic chemicals, concentrated/fuming acids and organic
solvents. (4 hrs)

Language of analytical chemistry: Definitions of analysis, determination, measurement,

techniques and methods. Significant figures, Classification of analytical techniques. Choice of an
analytical method. Errors and treatment of analytical data: Limitations of analytical methods –
Errors: Determinate and indeterminate errors, some important terms replicate, outlier, Accuracy,
precision, ways of expressing accuracy, absolute error, relative error, minimization of errors.
Statistical treatment of random errors, mean, median, range, standard deviation and variance.
External standard calibration. Numerical problems.
(6 hrs)

Titrimetric analysis: Basic principle of titrimetric analysis. Classification, preparation and

dilution of reagents/solutions. Equivalent masses of compounds Normality, Molarity and Mole
fraction. Use of N1V1= N2V2 formula, preparation of ppm level solutions from source materials
(salts), conversion factors. Numerical problems. (2hrs)

Acid-base titrimetry: Titration curves for strong acid vs. strong base, weak acid vs. strong base
and weak base vs. strong acid titrations. Titration curves, quantitative applications – selecting and
standardizing a titrant, inorganic analysis - alkalinity, acidity. (2hrs)

Introduction:

Everything in this universe, from subatomic particles to planets is made up of matter. Analytical
chemistry deals with obtaining information about composition and structure of matter. It is a
study of methods used to separate, identify and quantify matter. Though separation, a sample of
matter (analyte) is isolated. Identification of analyte is done through qualitative analysis. The
quantification determines the numerical amount or concentration through quantitative analysis.
Basic laboratory practices:

[Note: There are various terms like “basic laboratory practices,“safety in chemical laboratory”,
“rules of working in analytical laboratory” “precautions to be taken while handling toxic
chemicals”, etc., in the above syllabus. These terms are synonymous in their meaning and
generally regarded as safe laboratory practices. These are practices or rules of work to be
followed in any laboratory, chemical, analytical or otherwise. Hence the study material is written
for this part to include the concepts of all the above termsunder one common heading as “basic
laboratory practices”]
All analytical chemistry operations are carried out inside a specially designated space called a
laboratory. There are certain general or basic laboratory practices which should be followed by
every student, teacher or researcher in a laboratory. These are as follows:
1. Before entering a lab, it is absolutely essential to wear proper laboratory attire. These
include lab coat, hand gloves, eye protection goggles, face masks and PPE (if
necessary).
2. Consuming food in a lab is prohibited. Eating and drinking can increase the risk of
exposure to chemicals. It can also cause contamination of experiments and attract
pests.
3. It is important to maintain good hygiene in the lab. This involves washing hands
after handling chemicals, keeping personal items separate from lab work, not
applying cosmetics while in the lab, etc
 4. Every chemical and item of glassware used should be entered in a register so that it
becomes easier to check the stock at regular intervals.
5. Recording of observations, data collected, calculations, etc are imperative tosuccess
in lab work.
6. The person working in a lab should know how to use storage containers properly.
Chemicals should be stored in containers made of materials which will not react with
them. Highly inflammable chemicals in large volumes should be stored in fireproof
cabinets Acids should be placed in fuming chambers. Proper storage of batteries and
their disposal after use is also of crucial importance.
7. In a laboratory, labelling the work space is important. All containers should be
labelled with their contents so that anyone visiting the lab will know what hazards
may be present. Areas of risk should also be notified with proper labels.
8. One should never work alone in a laboratory setting. Having other researchers
around will help in noticing hazards and provide faster support in the event of an
emergency. It is important to notify your supervisor or teacher before entering the
lab and upon departure.
9. All lab members should be familiar with the use of safety equipment such as fire
extinguishers, first aid kit, etc. Calibration of volumetric glassware:

Calibration is a process of comparison between a known measurement standard for an instrument

and the observed measurement using that instrument. Calibration of measuring instruments has
following objectives:
1. It checks the accuracy of the instrument.
2. It determines the traceability of the measurement
3. It helps in repairing of the instrument which is out of calibration.

Calibration of pipette, standard flask and burette:

The general procedure is based on measuring the mass of water either contained or
delivered by the apparatus. This mass is then converted to volume by using the density of water.
Then the difference between expected volume and observed volume is found.

Note: For a detailed procedure about calibrating above glassware items refer to practical
 manual of practical record

Sampling:
Sampling is an integral step of chemical analyses. According to IUPAC, a sample is
defined as a portion of material selected from a large quantity of that material. Meaningful
analytical results can only be achieved if the test sample is completely representative of the
material to be analysed. Hence sampling is the process of collecting a smaller portion of a material
which is present in larger quantity such that the sample is representative of the larger quantity.
.The analyst must therefore know about the normal standard sampling procedures employed for
different type of materials.

A sample can be a solid, liquid or gas. It can be homogeneous or heterogeneous If it ishomogeneous

in nature then sampling will be easy since it can be collected as it is and used. But if it is a
homogenous, then it is necessary to combine a number of portions to ensure that arepresentative
sample is finally selected for analysis.

In most cases of chemical analyses, the sample collected may not be suitable for directanalysis.
Hence it must be converted to proper form which yields to proper analysis. The technique of
preparing a proper sample for analysis is called sample preparation.

If the analyte is a homogeneous liquid then sampling will be easy. But if it is a solid mixture,
then it is necessary to combine a number of portions to ensure that a representative sample is
finally selected for analysis,
In general, for solids, the sampling methods used are:

1. Solid–liquid extraction: In this method, a solvent which is able to dissolve the analyte is
added to the solid. The remaining insoluble particles are separated from the dissolved
fraction by filtration, decantation or centrifugation.
2. Soxhlet extraction: In this case the sample is placed in a disposable porouscontainer
known as a ‘thimble’. By continuous refluxing, an organic solvent is madeto through the
thimble and it dissolves the analyte molecules.
3. Ultrasonic radiation method: The finely divided sample is immersed in an ultrasonic
 bath containing a suitable solvent. Ultrasonic radiation is passed by meansof a probe which
gives the required energy to the sample molecules to dissolve in the solvent.
For liquids, the following sampling methods are used:

1. Solid phase extraction: Here the liquid is passed through a solid phase, which
selectively retains the analyte. Thereafter, the analyte can be eluted with another strong
solvent.
2. Liqui-liquid extraction: In this method, the sample is partitioned between two
immiscible phases. The extraction solvent and extraction conditions are chosen in such
a way that maximum difference in solubility is obtained.
3. Evaporation method: Here the liquid is removed by gentle heating at atmospheric
pressure with flowing air or inert gas in a rotary evaporator.
Weighing:

Weighing is of utmost importance in analytical chemistry work because it is essential to knowthe

amount of reagents to be used for preparing reagent solutions, for calibration of volumetric
glassware and also to use exact amounts of reactants during chemical syntheses. The expected
yield of a product during its chemical synthesis depends on the initial weight of the reactant since
there is a direct link between the number of moles and the number of molecules reacting.

In an analytical laboratory, all weighing is done using analytical balances which canrecord the
weight sensitively up to the fourth decimal place.

Chemicals are never weighed directly on the pan of a laboratory balance. Instead the mass is
determined by a process known as “weighing by difference’. A suitable container, which is
usually a small weighing bottle or a small beaker, is weighed empty first. The desiredchemical is
added to the container and the total mass of the combination is determined. Then by subtracting
the mass of the container from the total mass of container plus the chemical, the mass of the
chemical is found.
Following rules are important while weighing:

1. Before weighing make sure that the balance is calibrated and the pan is clean
2. Do not handle objects to be weighed with bare hands. Use tongs, gloves etc.
 3. Do not weigh hot or cold objects in the balance. Hot objects will lower the readings dueto
air byuancy and cold objects will give higher readings due to condensation of water
vapour
4. Never spill chemicals inside the analytical balance enclosure.
5. Do not overload the balance. Weigh objects which are below the general limit of the
balance.
Drying:

During lab work it is necessary to dry glassware, analytical standard compounds, synthesized
compounds, crucibles used in gravimetry and solvents. The term ‘drying’ generallymeans removal
of water from an object, usually a chemical, using the process of evaporation.
A solid sample can be dried by a variety of methods;

1. Heating the chemical

2. Using a drying agent in a closed container to absorb the solvent
3. Reducing the atmospheric pressure
4. Drying in electronic ovens
5. Drying in dessicators filled with a dessicant (drying agent) like CaCl2 at the bottom

Drying a liquid means removal of water from a liquid chemical or a solution of chemical in a
water-immiscible solvent. A solid drying agent like MgSO4, Na2SO4, CaCl2, etcis added to the
liquid or solution. The drying agent should be totally insoluble in the liquid, should not react
with it, absorb water quickly and be easily filterable.

Dissolving:

Before the analysis of a sample, the sample should be dissolved so that either the analyte passes
in to the solution leaving behind unwanted impurities or the impurities themselves passin to
solution so that analyte alone can be recovered.
Following methods are used to dissolve inorganic solids:

1. Acid treatment: Strong mineral acids are used to dissolve solids. Hydrochloric acid isa
general solvent for most metals. Nitric acid being a strong oxidizing agent will dissolve
 non-ferrous alloys and sulphides, Perchloric acid is also a good solvent for metals.
2. Fusion technique: This technique is used for inorganic materials which do not dissolvein
acids. Here the sample is mixed with an acidic or basic flux (eg:sodium carbonate) in the
ratio 1:10 and the mixture is heated in a crucible till the flux melts and becomes clear.
During this process, the insoluble materials in the sample react with the flux to form a
soluble product. The product is then cooled and the cooled solid is dissolved in a
suitable dilute acid or water.

General rules for performing quantitative determinations (volumetric and gravimetric):

Volumetric analysis is a widely-used quantitative analytical method. As the name implies,this

method involves the measurement of volume of a solution of known concentration whichis used
to determine the concentration of the analyte.
General rules for performing volumetric analysis are as follows:

1. Prepare a solution from an accurately weighed sample of the material to be analyzed.

2. Choose a substance that will react rapidly and completely with the analyte and
prepare a standard solution of this substance.
3. Place the standard solution in a burette and add it slowly to the unknown. This process
is called titration and the solution in the burette is called the titrant. Continue the
titration until the reaction is complete; that is, until the amount of reactant added is
exactly the amount required to react with all the constituent being analyzed. Thispoint is
called the equivalence point, and can be detected by adding an indicator to the unknown
solution before beginning the titration. An indicator is a substance thatgives a color change at
or near the equivalence point. The point at which the color change occurs is the end point of
the titration.
4. Measure the exact volume of standard solution required from burette readings before
and after the titration. Since the molarity of the standard solution is known, the
number of moles of titrant can be calculated. From a knowledge of the equation for
the reaction, the number of moles of constituent present in the sample can also be
calculated.
Gravimetric analysis is a technique through which the amount of an analyte can be determined
through the measurement of mass. Here the analyte is isolated in the form of a precipitate in
 pure state by adding a suitable precipitating agent. The precipitate is dried and weighed. From
the formula weight of the precipitate the weight of the analyte in the isolated precipitate is
calculated
General rules for performing gravimetric analysis are as follows:

1. Weigh a certain amount of the analyte in to beaker

2. Dissolve the analyte in a suitable solvent
3. Add a precipitating agent to the solution
4. Test for completion of precipitation by adding a drop of the precipitating agent and
looking for any sign of fresh precipitation
5. Filter the precipitate by vacuum filtration
6. Dry and weigh the precipitate
7. Use stoichiometry to determine the mass of the analyte

Rules of fire prevention and accidents:

Fire prevention requires understanding the properties of flammable chemicals along with
following other safe laboratory work procedures. Rules for fire prevention and accidents are given
below:
1. All flammable chemical should be properly labeled and safety data sheets should be
provided for them
2. Keep containers closed except when in use
3. Keep the flammable materials away from heat, sparks and sources of ignition
4. Maintain adequate ventilation to remove routine vapor emissions. A laboratory hood isa
must for this purpose
5. Minimize the quantity of flammable chemicals in work area
6. Installation of fire extinguishers and knowledge of using them is absolutely necessary

First Aid:

Accidents do not often happen in well-equipped chemistry laboratories if students understandsafe

laboratory procedures and are careful in following them. When an occasional accident does occur,
it is likely to be a minor one. The following information will be helpful to if an accident occurs:
1. A First Aid Box containing items like bandages , burn cream, antiseptic swipes, eye pad
dressing, etc should be placed in the laboratory
2. Burns – hold the affected skin under a stream of running water for at least 10 – 15 min.
Keep the wound open and apply burn ointment in case of minor burns. In case of strong
acid burns after washing with water rinse with dilute ammonia (1 – 2%) or sodium
bicarbonate solution.
3. Eyes – if corrosive liquid gets splashed into the eyes wash the eyes thoroughly with fresh
water. Seek medical help to rule out eye injury
4. Poisons – dilute the stomach contents by making the person drink 1 – 2 glasses of water
and try to induce vomiting before specific expert medical help can be provided.

Precautions to be taken while handling toxic chemicals, concentrated/fuming acids and

organic solvents:
Toxic chemicals:

1. Wear the appropriate PPE when handling toxic chemicals

2. Use the least hazardous chemical option for the task at hand and prepare only the
amount that is absolutely necessary for completing the job.
3. Wash work clothes separately from street clothes, or wear disposable clothing.
4. Wash your body thoroughly after using chemicals and before eating, drinking or using
the washroom.

Concentrated/fuming acids:

1. Work with acids and bases in well-ventilated areas or with devices thatremove the hazard,
such as local exhaust ventilation, fume hoods or similar capture devices.
2. Always add acid to water to prevent splattering
3. Clean-up spills promptly with appropriate materials.

Organic solvents:

1. Keeps tight lid on the container of the organic solvent whether it’s in useor not
2. Avoid inhaling the vapors of the solvent by keeping it away from you
 3. Avoid direct skin contact
4. Keep away plastic items like pens, key chains, spectacle frames from organic solvents

Language of Analytical Chemistry

Analytical chemistry methods refer to techniques used for the detection, identification,
characterization and quantification of chemical compounds.

Analysis - provides chemical or physical information about a sample. In an analysis we determine

the identity, the concentration, or the properties of an analyte. To make this determination we
measure one or more of the analytes chemical or physical properties

Analyte - The component in the sample of interest to us

Matrix - the remainder of the sample

Replicate - Replicate or replicate samples are portions of a material of approximately the same
size that are carried through an analytical procedure at thesame time and in the same way.

The terms analyze and determine have two different meanings. We say a sampleanalyzed for part
or all of its constituents. The substances measured are called theanalytes. The process of measuring
the analyte is called a determination.

The major types of analytical techniques used in chemistry determine the chemicalcomposition of
sample qualitatively and quantitatively.

A technique is any chemical or physical principle we can use to study an analyte.

A method is the application of a technique for a specific analyte in a specificmatrix
A procedure is a set of written directions telling us how to apply a method to aparticular sample,
including information on obtaining samples, handling interferents, and validating results
Analytical technique is a method that is used to determine a chemical or physical property of a
chemical substance, chemical element, or mixture. There area wide variety of techniques used for
analysis, from simple weighing to advanced techniques using highly specialized instrumentation.

The two main methods used are wet chemistry or classical method and instrumentmethods

Classical methods
The majority of the classical analytical methods rely on chemical reactions toperform an analysis.
Classical qualitative analysis is performed by adding one or a series of chemical reagents to the
analyte. By observing the chemical reactions and their products, one can deduce the identity of the
analyte. The added reagents are chosenso that they selectively react with one or a single class of
chemical compounds to form a distinctive reaction product.

Classical quantitative analysis can be divided into

1. Volumetric analysis
2. Gravimetric analysis

1. Titrimetric or Volumetric: technique to determine the concentration of the sample. The

substance is allowed to react with a suitable reagent taken as a standard solution and the
volume of the solution required for complete reaction is determined eg: acid-base,
redox,complex , precipitation reactions
2. Gravimetric: substance being determined is converted into an insolubleprecipitate which
is collected and weighed

Instrumental methods
They are based on the measurement of a physical property of the sample, for example, an electrical
property or the absorption of electromagnetic radiation.Instrumental techniques are generally more
sensitive and selective than the classical techniques but are less precise. They are usually more
rapid, may be automated, and may be capable of measuring more than one analyte at a time.
The instrumental methods of chemical analysis are divided into categories according to the
property of the analyte that is to be measured. Many of the methods can be used for both qualitative
and quantitative analysis. The major categories of instrumental methods are the spectral,
electroanalytical, and separatory
Electroanalytical Techniques:
Involves the measurement of such electrical properties as voltage, current, in anelectrochemical
cell containing the analyte. The four main categories are
1. Potentiometry- difference in electrode potential is measured
2. Coulometry – transferred charge is measured over time
3. Amperometry- cell’s current is measured over time
4. Voltammetry - cell’s current is measured while actively altering the cell’spotential
5. Electrogravimetry – electrolysis is carried and substance deposited on one of the
electrodes is measured

Spectroscopic Techniques:
These are based on the measurement of the interaction between electromagnetic radiation and
analyte atoms or molecules.
There are different applications such as Ultraviolet-visible spectroscopy, IR spectroscopy, Raman
spectroscopy, NMR spectroscopy, X-ray method, Atomic absorption spectroscopy, Atomic
emission spectroscopy, etc.,

Thermal analysis:
Based on weight measurements or energy changes as a function of temperature- Eg:
Thermogravimetric analysis (TG), Differential therml analysis (DTA). calorimetry etc.,

Radiochemical methods:
The spontaneous emissions of particles or electromagnetic radiation from unstable atomic nuclei
are monitored. The intensity of the emitted particles or electromagnetic radiation is used for
quantitative analysis, and the energy of the emissions is used for qualitative analysis

Separatory methods:
Chromatography and mass spectrometry are two such methods that are particularlyimportant for
chemical analysis
a) Chromatography techniques are particularly powerful for analyzing complex mixtures..
 Constituents are separated as they are pushed through (eluted from) a columnof appropriate
material that interacts with the analytes to varying degrees, and these aresensed with an
appropriate detector as they emerge from the column, to give a transient peak signal,
proportional to the amount of each.
b) Mass spectrometry: Measures mass to charge ratio of molecules using electric and
magnetic fields.

The advantages of instrumental methods over classical methods include:

1. The ability to perform trace analysis
2. Generally, large numbers of samples may be analyzed very quickly.
3. Many instrumental methods can be automated.
4. Most instrumental methods are multi-channel techniques
5. Less skill and training is usually required

Choice of / Selecting an Analytical Method

In order to select an analytical method, it is essential to define clearly the nature ofthe analytical
problem. In general, the following points should be considered whenchoosing an instrument for
any measurement.
Accuracy and precision required Available sample amount Concentration range of the analyte
Interference in sample
Physical and chemical properties of the sample matrixNumber of sample to be analyzed
Speed, ease, skill and cost of analysis

Limitations of Analytical Methods.

1. The function of the analyst is to obtain a result as near to the true value aspossible by the
correct application of the analytical procedure employed
2. The level of confidence in the results will be very small unless there is a knowledge of the
accuracy and precision of the method used as well as beingaware of the sources of error in
the measurement.
Outliers
Every experimentalist is familiar with the situation in which one (or possible more) of a set of
results appears to differ unreasonably from the other in the set.Such a measurement is called an
outlier.
For example if the following results were given for a titration:12.12, 12.15, 12.13, 13.14, 12.12 ml

Precision
1. A measure of how closely individual measurements agree with one another.
2. It describes the reproducibility of measurements.
3. Precision is readily determined by simply repeating the measurement on replicatesamples.
4. Precision of a set of replicate data may be expressed as standard deviation,variance, and
coefficient of variation.
Accuracy:
 Refers to how closely individual measurements agree with the correct or trueor accepted
value. It is expressed by the error.
 Accuracy is often more difficult to determine because the true value isusually unknown.
An accepted value must be used instead.

Examples
Let us understand this concept using an experiment, suppose the true mass for a beaker is 3.5g and
an analyst takes two measurements in an experiment and reportsthe masses as 3.45g and 3.46g for
the same beaker. This reported values are precisebut not accurate.

Accuracy expresses correctness of a measurement whereas Precision the reproducibility of a

measurement. Precision always accompanies accuracybut a high degree of precision does not
imply accuracy.

Eg: The actual concentration is 0.1122 N

Student-1 is getting – Accurate and Precise
0.1122 0.1121 0.1122 0.1122 0.1122N
Student – 2 is getting – Precise but not accurate
 0.1126 0.1125 0.1126 0.1126 0.1126 N

Error often denotes the estimated uncertainty in a measurement or experiment.

Ways of expressing Accuracy

Accuracy is expressed in terms of either absolute or relative error.

Absolute Error
The absolute error of a measurement is the difference between the measuredvalue and the true or
known or theoretical value.
If the measurement result is low, the sign is negative; if the measurement result ishigh, the sign is
positive.
Error =Theoretical value – Experimental value

Relative Error
The relative error of a measurement is the absolute error divided by the true value. Relative error
may be expressed in percent, parts per thousand, or parts per million,depending on the magnitude
of the result

Measurement of Errors
All measurements have errors associated with them. These errors fall into twocategories:
1. Determinate errors or Systematic errors – affect the accuracy of themeasurement,
or the closeness of the result to the “true” value;
2. Indeterminate errors or Random errors– affect the precision of themeasurements, or
the closeness of the results to each other
 Determinate errors
 Have a definite value,
 An assignable cause, and
 Are of the same magnitude for replicate measurements made in the sameway.
 They lead to bias in measurement results.

There are three types of determinate errors:

 Instrumental errors.
 Method errors
 Personal errors

a. Instrumental Errors
o These are caused by nonideal instrument behavior, by faulty calibrations, or by use under
inappropriate conditions
o Pipettes, burettes, and volumetric flasks may hold or deliver volumes slightlydifferent from
those indicated by their graduations.
o Calibration eliminates most systematic errors of this type.
o Electronic instruments can be influenced by noise, temperature, pH and are alsosubject to
systematic errors.
o Errors of these types usually are detectable and correctable

b. Method Errors
o The nonideal chemical or physical behavior of the reagents and reactions on which an
analysis is based often introduce systematic method errors.
o Such sources of nonideality include the slowness of some reactions, the incompleteness of
others, the instability of some species, the lack of specificity of most reagents, and the
possible occurrence of side reactions that interfere with themeasurement process.
o Errors inherent in a method are often difficult to detect and hence, these errors areusually
the most difficult to identify and correct.
o
c. Personal Errors
Result from the carelessness, inattention, or personal limitations of theexperimenter.
Many measurements require personal judgments.
Examples include estimating the position of a pointer between two scale divisionsthe colour of a
solution at the end point in a titration, or the level of a liquid with respect to a graduation in a
pipette or burette.

 Judgments of this type are often subject to systematic, unidirectional errors

 A universal source of personal error is prejudice, or bias.
 Number bias is another source of personal error that varies considerably fromperson to
person.
 The most frequent number bias encountered in estimating the position of a needleon a scale
involves a preference for the digits 0 and 5.
 Also common is a prejudice favoring small digits over large and even numbersover odd.
 Digital and computer displays on pH meters, laboratory balances, and other electronic
instruments eliminate number bias because no judgment is involved intaking a reading.

Elimination of Systematic (Instrumental and Personal) Error

1. Periodic calibration of equipment is always desirable because the response of most
instruments changes with time as a result of component aging, corrosion,or mistreatment.
2. Most personal errors can be minimized by careful, disciplined laboratory work.
3. It is a good habit to check instrument readings, notebook entries, andcalculations
systematically.
 4. Errors due to limitations of the experimenter can usually be avoided by carefullychoosing
the analytical method or using an automated procedure.
5. Running a blank determination.
6. Running a parallel determination

Indeterminate or Random Errors

 Random errors are always present during any measurement and can’t becontrolled.
 They arise from minor differences in sampling between different people, from how
people read a burette or interpret an endpoint color, and even fromelectrical noise.
 The contribution of random error to a measurement can be estimated througherror
propagation methods, by measuring the standard deviation of a series of measurements
that show a normal distribution, and by monitoring the behavior of a piece of equipment
over time.

In general, the magnitude of systematic errors greatly exceeds those of random errors,so that is
where analysts typically focus their attention to minimize overall error in a process.

Significant figures:
It is a digit (one of the ten numerals: 0,1 ,2,3….9) which denotes the quantity inthe place where it
stands
They are meaningful digits which are known with certainty.

Note:
The leading zeros are zeros occurring to the right of the decimal point which are not
considered as significant zeros.
The captive zeros are zeros between the non-zero digits; these captive zeros aresignificant.
The trailing zeros are zeros that are occurring at the end and are not significant.

Rules for determining the number of significant figures:

1. All non-zero digits are significant.
For example: - 359.2 cm - there are 4 significant figures
6575 cm there are four significant figures
 0.543 there are three significant figures
2. Zeros preceding to first non-zero digit are not significant.
For example: - 0.05 has one significant figure
0.0012 has two significant figures.
3. Zeros between two non-zero digits are significant.
For example: - 7.001 have four significant figures.
4.5006 have five significant figures
4. Zeros at the end or right of a number are significant provided they are on the rightside
of the decimal point.
For example: - 6.00 g has three significant figures.
6.00s only one significant figure.
0.500 has three significant figures.0.00500 – 3 significant figures
5. Scientific Notation
Scientific notation form: a x 10b, where “a” and “b” are integers, and "a" has to bebetween 1 and
10.

Disregard the “10b,” and determine the significant digits in “a.”

Eg: 5748 - The scientific notation for 5748 is 5.748 x 103

It has 4 significant digits.
b) 152 x 106 has 3 significant digits.
c) 14000 It can be written as 1.4 x 104 and has 3 significant figures
6. Exact numbers have an infinite number of significant figures.
For example:- In 3 balls or 40 eggs, there are infinite significant figures asthese are exact
numbers and can be represented by writing infinite number of zeros after placing a decimal
i.e.,3 = 3.000000
or 40 = 40.000000
π=22/7=3.1428….. has infinite significant figures

Rules for Using Significant Figures

1. In multi-step calculations, you may round at each step or only at the end.
2. Exact numbers, such as integers, are treated as if they have an infinite numberof significant
figures.
 3. In calculations, round up if the first digit to be discarded is greater than 5 andround down
if it is below 5. If the first discarded digit is 5, then round up if a nonzero digit follows it,
round down if it is followed by a zero.

Addition & Subtraction

For addition and subtraction, the answer should have the same number of decimalplaces as the term
with the fewest decimal places
i.e.,the result cannot have more digits to the right of the decimal point thaneither of the original
numbers.
For example: -
a) (12.11 + 18.0+ 1.012) = 31.122. = 31.1
(As 18.0 have only one digit after the decimal point therefore the resultwill be 31.1, one digit after
the decimal point.)
b) Y = 232.234 + 0.27 Find Y.

Y = 232.50

12.793 + 4.58 + 3.25794 = 20.63094 = 20.63

(Since 4.58 has 2 decimal places, which is the least number of decimal places.)

Multiplication & Division

For multiplication and division, the answer should have the same number ofsignificant figures as
the term with the fewest number of significant figures.

(Note: significant numbers are not considered in the final result likemultiplication or division
operation.)
a) 2.5×1.25 = 3.125 = 3.1
(Because 2.5 have two significant figures and the result should not havemore than two significant
figures.)
b) 56.937/0.46 = 130.29782609 =1.3 x 102
Since 0.46 has only 2 significant figures. Notice that 130 would be ambiguous, so
scientific notation is necessary in this situation.)
Mean
The mean, also called the arithmetic mean or the average.
It is obtained by dividing the sum of replicate measurements by the number ofmeasurements in the
set:

Median
is the middle value in a set of values.
If you have a set of N values and put them in either ascending or descending order.
For an odd value of N, the center number in the series is the median value.
For an even value of N, it is the average of the two central numbers.
The median is denoted by M and may be used as an estimate of the central value.
Standard Deviation
The variance is found by summing the squares of the deviations and dividing thatsum by n–1 (since
it is a sample instead of a population).
The square root of the variance gives a kind of average of the deviations from themean, which is
called a sample standard deviation.
It is denoted by the letter s.

Calculation of Standard Deviation

1. Calculate the mean of the numbers.
2. Find the deviations from the mean.
3. Square each deviation.
4. Sum the squared deviations.
5. Divide the sum in Step 4 by (n – 1)
6. Take the square root of the quotient in Step 5.

Range
The range of a set of measurements is the difference between the largest andsmallest values in the
data set.
Its major advantage is the ease with which it can be computed.It is very sensitive to the smallest
and largest data values
Its major shortcoming is its failure to provide information on the dispersion of thevalues between
the two end points.
Range = largest value - smallest value
Variance
Variance is a numerical value that shows how widely the individual figures in a setof data distribute
themselves about the mean. That is how far each number is from the mean, and thus from each
other.
A variance of zero value means all the data are identical.
Greater the variance, more are the values spread out about mean, hence from eachother.
Less the variance, less are the values spread out about mean, hence from each other,and variance
can’t be negative.
The main difference between population variance and sample variance relates tocalculation of
variance.
When the variance is to be calculated from population data, n is equal to the numberof items
When the variance is calculated from sample data 1 is to be deducted from n beforedividing the
sum of the squared deviations.
Thus, if sample data have 100 items, the denominator would be 100 – 1 = 99.

The value of variance calculated from sample data is higher than population data.

Sample variance (s2) is a measure of the degree to which the numbers in a list are spread out.
If the numbers in a list are all close to the expected values, the variance will be small. If they are
far away, the variance will be large.

Sample variance s2 = ∑ (x − x̅ )2 / n – 1

Steps:
1. Find the mean of the data.
2. Subtract the mean from each data point.
3. Take the summation of the squares of values obtained in the previous step.
4. Divide this value by n - 1.

Population variance σ2 = ∑ (x − x̅ )2 / n
The following data were collected as part of a quality control study for the analysisof sodium
in serum; results are concentrations of Na+ in mmol/L.
140, 143, 141, 137 ,132, 157, 143, 149, 118, 145 .
Report the mean, the median, the range, the standard deviation, and the variance for this data.
Mean = 140 +143+141+137+132+157+ 143+ 149+118+145 = 1405
10
=140.5 mmol/L

Median:
118 132 137 140 141 143 143 145 149 157
5 th value + 6th value = 142 mmol/L
2 2
Range: = Highest – Lowest = 157 – 118 = 39 mmol/L

Standard deviation

X x- xi (x- xi)2
140 -0.5 0.25
143 2.5 6.25
141 0.5 0.25
137 -3.5 12.25
132 -8.5 72.25
157 -16.5 272.25
143 2.5 6.25
149 8.5 72.25
118 -22.5 506.25
145 4.5 20.25

968.5

=√ 968.5/9 = 10.37
Sample variance s2 = ∑ (x − x̅ )2 / n – 1
= 968.5/9
= 107.173

External standard calibration

External standard calibration is one of the most common approaches to calibrationsSimplest
and most common form of calibration
An external standard is like the internal standard (known behaviour), but is not added to the
unknown. Rather it is run alone, as a sample, and usually at differentconcentrations, so you can
generate a standard curve.
For an external standard quantitation, known data from a calibration standard and unknown
data from the sample are combined to generate a quantitative report. It iscalled external standard
because the standard or known material is separate or external to the unknown material.

 It involves a simple comparison of instrument responses from the sample to theresponses

from the target compounds in the calibration standards.
 Sample peak areas (or peak heights) are compared to peak areas (or heights) of thestandards.
 The ratio of the detector response to the amount (mass) of analyte in the calibration
standard is defined as the calibration factor
Equation for Calibration Factor (External Standard Curve) CF = (Ax) / (Cx)Where: Ax = Area
of the compound
Cx = Concentration of the compound
Steps:
Prepare samples known quantities of analyte over a relevant range including blanksControls for
sample preparation/matrix should be used, matched to the unknown samples
Carry out and record measurements
Plot quantity/concentration of analyte vs. response
Linear regression with least squares analysis is used to determine response(expressed as y =
bx+a)
Repeat as and when appropriate (when it is likely that an unacceptable drift willhave occurred)
Again, the peak areas are related to the known amounts of external standard run.
Advantages
External standards do not correct for losses that may occur during preparation ofthe sample,
such as extraction, centrifugation, evaporation, etc.
This ‘external standard’ calibration model will work only when the instrument reproducibly
introduces fixed volume aliquots of sample into the system and thereis no mechanism by which
the response of one sample specie can influence the instrument response with respect to another
 May only need one calibration plot (of 5-10 samples) for 10’s to 100’s of unknownsamples
Can be easily automated
It can be applied to a wide variety of methods
Simple statistics will provide estimates of uncertainty for the method

Disadvantages
Requires care to match conditions and matrix to that of the unknown samplesDoes not control
for sudden changes in method performance

TITRIMETRIC ANALYSIS

Basic Principles of Titrimetric analysis:

Titrimetric analyses are the quantitative analytical methods in which the amount of reagent
consumed by the analytes completely is measured using reagent of known concentration. The
solution whose concentration is known as standard solution. The weight of the substance to be
determined is calculated from the volume of the standard solution used and the chemical
equation and relative molecular masses of the reacting compounds.

Titrant: The reagent of known concentration in the titrimetric analysis. Eg: Sodium oxalate

Titrand: The substance being titrated. Eg: KMnO4

The titrant is filled in burette and titrand is taken in conical flask. The point of completion of
the reaction is called as equivalence point or end point which can be detected using indicators.
Indicators are of three types: Internal indicator (phenolphthalein, methyl orange,
diphenylamine etc), external indicator (potassium ferricyanide) and self-indicator (KMnO4).

Types of titrimetric analyses:

 Volumetric titrimetry is a type of analysis in which the measured quantity is the volume of a
standard reagent whose concentration.
 Coulometric titrimetry is a type of titrimetry in which the measured quantity is the charge in
coulombs required to complete a reaction with the analyte. In this method, constant direct
electrical current of known magnitude that consumes analyte is considered as reagent. The time
required andthe total charge to complete the electrochemical reaction are measured.
 Potentiometric titration: titration in which potential between an indicator electrode and a
 reference electrode is measured.
 Amperometric titration: Titration in which the current which passes through the titration cell
between an indicator electrode and a depolarised reference electrode at a suitable applied e.m.f.
is measured.

Reagents:
 The reagents used for quantitative analysis should be of analytical grade or purest
form.
 To reduce the errors, reagent bottles should not be opened for long time and should not
be returned to the reagent bottle.
 Liquid reagents should be poured directly from the bottle and pipette should not be
inserted into the bottle.
Standard solutions:
 Reagent solution is prepared usually by weighing out approximate amount of the
substance required using watch glass or plastic container and then required amount of
solvent is added after being measured using measuring cylinder.
 Standard solution is prepared using standard flask or volumetric flask. A known amount
of substance (which is calculated using chemical formula) is weighed accurately in
weighing bottle which is already weighed and transferred into the standard flask using
stemmed funnel. During the transfer, no chemical particles should be lost. The funnel is
thoroughly washed using deionised water inside and out. The contents in the flask will be
dissolved using either swirling or shaking. Then the solution will be made upto the mark
in the standard flask. DO NOT HEAT THE GRADUATED or STANDARD FLASK.
 Solutions which are not affected by air, moisture or light can be prepared and stored in 1
L pr 2.5 L bottles.
 Solutions which are already prepared can be diluted using the following formula:
N1V1 = N2V2
Where, N1 and V1 are the known normality and volume of the standard
solution; N2 and V2 are the normality and volume of the solutions to be
diluted.

Equivalent masses:

 The equivalent of a substance is the amount of that substance in a chemical reaction,

which combines with, releases or replaces that amount of hydrogen which is combined
 with 3 grams of carbon – 12 in methane.
 A normal solution is one in which one equivalent of a defined species per litre
according to the specified reaction. Particularly, the amount of hydrogen replaced by
one equivalent of any substance or one equivalent amount of electricity. The reaction
to which the definition is applied should be clearly mentioned.

Normality = number of equivalents / number of litres

= number of milliequivalents / number of millilitres

If the solution volumes of two different substances A and B which exactly react with one
another are VA mL and VB mL respectively, then these two contains same number of
milliequivalents of A and B respectively.
i.e., VA x normalityA = VB x normalityB

Molarity:
The number of moles of the component (of solute) present in one litre (1 dm3) of the solution.
Molarity =Moles of solute / Volume of solvent = n / V

Molality:
The number of moles of the component (or solute) present in one kilogram of the solvent.
Molality =Moles of solute / Mass of solvent = n / m

Mole Fraction:
In a solution containing n1 moles of component 1, n2 moles of component 2, n3 moles of
component 3… and so on, the mole fraction xi of the ith component is defined as,
xi= ni / ∑ni = ni / (n1 + n2 + n3…ni)
The sum of mole fractions of the components of a mixture (or a solution) is unity.

Parts per million (ppm):

The concentration of a component A in solution in parts per million (ppm) is defined as,
(ppm)A = (mass of solute / Total mass) X 106
 ACID-BASE TITRATION & CURVES

The Concept of a Weak/Strong Acid and Weak/Strong Base:

A strong acid dissociates (or ionizes) completely in aqueous solution to form hydronium ions
(H3O+)

A weak acid does not dissociate completely in aqueous solution to form hydronium ions
(H3O+)

A strong base dissociates completely in aqueous solution to form hydroxide ions (OH-)

A weak base does not dissociate completely in aqueous solution to form hydroxide ions (OH-
).

Examples of weak/strong acids and bases:

Type Examples

StrongAcids
hydrochloric acid (HCl), sulfuric acid (H2SO4), nitric acid (HNO3).

Weak Acids acetic acid (CH3COOH), hydrofluoric acid (HF), oxalic acid (COOH)2

StrongBases sodium hydroxide (NaOH), potassium hydroxide (KOH), lithium

hydroxide (LiOH)

Weak Bases ammonium hydroxide (NH4OH), ammonia (NH3)

Weak acids and weak bases always exist as conjugate acid-base pairs in an aqueous solution
as represented below
 Here, HA is the acid and A- is termed as the conjugate base of HA

In the above reaction, A- is a base and HA is the conjugate acid of A-.

Note: Weak acids have strong conjugate bases, while weak bases have strong conjugate acids.
Asshown in the above two reactions, if HA is a weak acid, then its conjugate base A- will be a
strongbase. Similarly, if A- is a weak base, then its conjugate acid HA will be a strong acid.

The Concept of Equivalence:

• Molecular weight of HCl: 36.5 g, NaOH: 40.0g, NaCl: 58.5g, & H2O is 18.0g respectively.
• 36.5 parts by weight of HCl are chemically equivalent to 40.0 parts by the weight of NaOH.
• If to 36.5 g HCl, 100 g NaOH is added, only 40.0g alkali will react (which is chemically
equivalent to 36.5 g HCl) and rest 60.0g will remain unreacted. This is the principle of
CHEMICAL EQUIVALENCE
 Titration

What is ‘titration’?

Titration is a technique to determine the concentration of an unknown solution. In this process,

asolution of known concentration (titrant) is used to determine the concentration of an unknown
solution (titrand or analyte).
Typically, the titrant (the solution of known concentration) is added through a burette toa
known volume of the analyte (the solution of unknown concentration) until the reaction is
complete. Knowing the volume of titrant added allows us to determine the concentration of the
unknown analyte. Often, an indicator is used to signal the end ofthe reaction, the endpoint.

Terminologies Used in Titration/Titration Curves:

If you wish to find the concentration of an acid solution, you would titrate the acid solution
with asolution of a base (alkali) of known concentration – alkalimetry.
You titrate a base of unknown concentration with an acid of known concentration –
acidimetry.

Titrant: solution of a known concentration, which is added to another solution whose

concentration,has to be determined.

Titrand or analyte: the solution whose concentration has to be determined.

Equivalence point: point in titration at which the amount of titrant added is just enough to
completely neutralize the analyte solution. At the equivalence point in an acid-base titration,
moles of base = moles of acid and the solution only contains salt and water.

Indicator/Indicators:
The substance which indicates the end point of Titration by color change is called as an
indicator.

Properties of an indicator:
 Must have an easily observed colour change.
 Must change easily in the required pH range over the addition of ‘half’ a drop of
reagent
Titration curve
What is a titration curve?
 A titration curve is the plot of the pH of the analyte solution versus the volume of thetitrant
added as the titration progresses.

Types of Acid base Titrations

1. Titration of Strong acid – strong base

2. Titration of Weak acid – strong base

3. Titration of Strong acid – weak base

4. Titration of Weak acid- weak base

1. TITRATION OF A STRONG ACID WITH A STRONG BASE

(Strong- Strong Titration) (Running alkali into acid)
 Point 1: No NaOH added yet, so the pH of the analyte is low (it predominantly contains H3O+
from dissociation of HCl).

As NaOH is added dropwise, H3O+ slowly starts getting consumed by OH- superscript
producedby dissociation of NaOH. Analyte is still acidic due to predominance of H3O+.

Point 2: This is the pH recorded at a time point just before complete neutralization takes place.

Point 3: This is the equivalence point (halfway up the steep curve). At this point, moles of
NaOH added = moles of HCl in the analyte. At this point, H3O+ are completely neutralized by
OH- ions. The solution only has salt (NaCl) and water and therefore the pH is neutral i.e. pH =
7.

Point 4: Addition of NaOH continues, pH starts becoming basic because HCl has been
completely neutralized and now excess of OH- ions are present in the solution (from
dissociationof NaOH).

1) Titration of a weak acid with a strong base (Running alkali into acid):
Let’s assume our analyte is acetic acid CH3COOH (weak acid) and the titrant is sodium
hydroxide NaOH (strong base). If we start plotting the pH of the analyte against the
volume of NaOH that we are adding from the burette, we will get a titration curve as shown
below.
Point 1: No NaOH added yet, so the pH of the analyte is low (it predominantly contains H3O+
from dissociation of CH3COOH). But acetic acid is a weak acid, so the starting pH is higher
thanwhat we noticed in case 1 where we had a strong acid (HCl).
As NaOH is added dropwise, H3O+ superscript slowly starts getting consumed by OH-
(produced by dissociation of NaOH). But analyte is still acidic due to predominance of H3O+.

Point 2: This is the pH recorded at a time point just before complete neutralization takes place.

Point 3: This is the equivalence point (halfway up the steep curve). At this point, moles of
NaOH added = moles of CH3COOH in the analyte. The H3O+ ions are completely neutralized
by OH- ions. The solution contains only CH3COONa salt and H2O.

At the equivalence point the solution contains CH3COONa salt. This dissociates into acetate
ionsCH3COO- and sodium ions Na+. As you will recall from the discussion of strong/ weak
acids in the case 1, CH3COO- is conjugate base of the weak acid CH3COOH. So, CH3COO- is
relatively a strong base (weak acid has a strong conjugate base), and will thus react with H2O
to produce hydroxide ions (OH-) thus increasing the pH to ~ 9 at the equivalence point.
 Point 4: Beyond the equivalence point (when sodium hydroxide is in excess) the curveis
identical to HCl-NaOH titration curve (1) as shown below.

2) Titration of a strong acid with a weak base (Running alkali into acid):

Suppose our analyte is hydrochloric acid HCl (strong acid) and the titrant is ammonia NH3
(weakbase). If we start plotting the pH of the analyte against the volume of NH3 that we are
adding from the burette, we will get a titration curve as shown below.
 Point 1: No NH3 added yet, so the pH of the analyte is low (it predominantly contains H3O
fromdissociation of HCl).

As NH3 is added dropwise, H3O+ slowly starts getting consumed by NH3. Analyte is still acidic
due to predominance of H3O+ ions.

Point 2: This is the pH recorded at a time point just before complete neutralization takes place.

Point 3: This is the equivalence point (halfway up the steep curve). At this point, moles of NH3
added = moles of HCl in the analyte. The H3O+ ions are completely neutralized by NH3. But
again, do you spot a difference here??? In the case of a weak base versusa strong acid, the pH
is not neutral at the equivalence point. The solution is in fact acidic (pH ~ 5.5) at the
equivalence point.

Point 4: After the equivalence point, NH3 addition continues and is in excess, so the pH
increases.NH3 is a weak base so the pH is above 7, but is lower than what we saw with a strong
base NaOH(case 1).

3) Titration of a weak base with a weak acid (Running acid into alkali):
The common example of this would be ethanoic acid (acetic acid) and ammonia.

It so happens that these two are both about equally weak - in that case, the equivalence point
is approximately pH 7.
Here our analyte is NH3 (weak base) and the titrant is acetic acid CH3COOH (weak acid). If
we start plotting the pH of the analyte against the volume of acetic acid that we are adding from
the burette, we will get a titration curve as shown below.

If you notice there isn’t any steep bit in this plot. There is just what we call a ‘point of inflexion’
at the equivalence point. Lack of any steep change in pH throughout the titration renders
titrationof a weak base versus a weak acid difficult, and not much information can be extracted
from sucha curve.

Quantitative Applications:
Although many quantitative applications of acid–base titrimetry have been replaced by other
analyticalmethods, there are several important applications that continue to be listed as standard
methods. In thissection we review the general application of acid–base titrimetry to the analysis
of inorganic and organic compounds, with an emphasis on selected applications in
environmental and clinical analysis.First, however, we discuss the selection and standardization
of acidic and basic titrants.

Selecting and Standardizing a Titrant:

Most common acid–base titrants are not readily available as primary standards and must be
standardized before they can be used in a quantitative analysis. Standardization is
accomplished by titrating a known amount of an appropriate acidic or basic primary standard.

The majority of titrations involving basic analytes, whether conducted in aque- ous or
nonaqueous solvents, use HCl, HClO4, or H2SO4 as the titrant. Solutions of these titrants are
usually prepared bydiluting a commercially available concentrated stock solution and are stable
for extended periods of time. Since the concentrations of concentrated acids are known only
approximately, the titrant’s concentration is determined by standardizing against one of the
primary standard weak bases.

The most common strong base for titrating acidic analytes in aqueous solutions is NaOH.

Inorganic Analysis:
Acid–base titrimetry is a standard method for the quantitative analysis of many inorganic acids
and bases. Standard solutions of NaOH can be used in the analysis of inorganic acids such as
H3PO4 or H3AsO4, whereas standard solutions of HCl can be used for the analysis of inorganic
bases such asNa2CO3.

Inorganic acids and bases too weak to be analyzed by an aqueous acid–base titration can be
analyzed byadjusting the solvent or by an indirect analysis.

Acid–base titrimetry continues to be listed as the standard method for the

de- termination of alkalinity, acidity:

Acidity is a measure of a solution’s capacity to react with a strong base (usually sodium
hydroxide,NaOH) to a predetermined pH value. This measurement is based on the total acidic
constituent of asolution (strong and weak acids, hydolyzing salts, etc.) It is possible to have
highly acidic water buthave moderate pH values. Likewise, the pH of a sample can be very low
but have a relatively low acidity. Acidity is similar to a buffer in that the higher the acidity, the
more neutralizer is needed tocounteract it.

Alkalinity is the measure of a solution’s capacity to react with a strong acid (usually sulfuric
acid H2SO4) to a predetermined pH. The alkalinity of a solution is usually made up of
carbonate, bicarbonate, and hydroxides. Similar to acidity, the higher the alkalinity is, the more
neutralizing agent is needed to counteract it.