ANALYTICAL CHEMISTRY
Analytical chemistry is the branch of chemistry that deals with the analysis of different
substances. Analytical chemistry involves the separation, identification, and the
quantification of matter. It involves the use of classical methods along with modern methods
involving the use of scientific instruments. There are four major areas of analytical
chemistry that are of importance in their application to diverse scientific disciplines. These
areas are spectroscopy, acid-base methods, potentiometry, and chromatography. Analytical
chemistry deals with the solving of qualitative and quantitative problems. Analytical
chemistry is the science of obtaining, processing, and communicating information about the
composition and structure of matter. In other words, it is the art and science of determining
what matter is and how much of it exists. Analytical chemistry involves the following
methods:
The process of separation isolates the required chemical species which is to be analysed
from a mixture.
The identification of the analyze substance is achieved via the method of qualitative analysis.
The concentration of the analyte in a given mixture can be determined with the method of
quantitative analysis.
Today, the field of analytical chemistry generally involves the use of modern, sophisticated
instruments. However, the principles upon which these instruments are built can be traced
to more traditional techniques.
Branches of Analytical Chemistry (CHEMICAL ANALYSIS)
Chemical analysis is a method of Separation, Identification and Quantification of different
components present in the given sample by using different analytical techniques.
Two sub-branches come under Chemical analysis namely quantitative analysis and
qualitative analysis which can be explained as follows. These two methods form the
backbone of many educational labs of analytical chemistry.
1. Qualitative Analysis
Quality means the standard or the feature of one substance. Hence, Qualitative analysis
method deals with the determination of the quality of a particular compound, irrespective of
its quantity or concentration. In simpler words, the qualitative analysis does not measure
the amount of the substance but measures the quality of that material. One of the best
examples of this type of method is the observation of a chemical reaction, whether there will
be a change in colour or not. The qualitative analysis method can be measured in different
ways such as Chemical tests, flame tests, etc. Several such tests are widely used in salt
analysis (identification of the cation & anion of inorganic salts).
2. Quantitative Analysis
Quantitative Analysis is a method of determining the absolute or relative quantity regarding
the concentration of one or more substances present in a sample or compound.
For example, take a sample of an unknown solid substance. The chemists first use
“qualitative” methods to identify what type of compound is present in the sample; then he
adopts the quantitative analysis procedure to determine the exact amount or the quantity of
the compound present in the sample. Some Quantitative analysis techniques
include Gravimetric Analysis and Volumetric analysis, Titrimetric analysis, Spectro
photometric Analysis etc. Quantitative analysis, is further classified into two types,
(a). Classical methods of analysis & (b). Instrumental methods of analysis
(a)._ Classical Methods / Wet chemical Analysis method
There exist many classical methods of checking for the presence or absence of a particular
compound in a given analyte. One such example is the acid test for gold. Another example of
a classical method for qualitative analysis is the Kastle-Meyer test which employs
phenolphthalein as an indicator to check for the presence of haemoglobin in the given
analyte. Flame tests can be used to check for the presence of specific elements in an analyte
by exposing it to a flame and observing the change in the colour of the flame.
Gravimetric analysis is a classical method of quantitative analysis, which can be used in
analytical chemistry to determine the amount of water in a hydrate by heating it and
calculating the weight of the water lost. One of the better known classical methods of
quantitative analysis is volumetric analysis (also known as titration). In the titration
method, a reactant is added to the analyte till an equivalence point is obtained.
The classical methods of analysis is usually relies on chemical reactions between the
material being analyzed (the analyte) and a reagent that is added to the analyte.
Ex: Gravimetric methods, Titrimetric methods, etc.,.
(b)._ INSTRUMENTAL METHODS
In contrast to Classical methods, Instrumental methods of analysis typically depends on the
measurement of a physical property of the particular analyte by using an instrument. It
involves the use of an instrument, other than a balance, to perform the analysis. In some
cases, the instrument is used to characterize a chemical reaction between the analyte and an
added reagent; in others, it is used to measure a property of the analyte. Mass Spectroscopy
involves the measurement of the interaction between electromagnetic radiation and the
atoms or molecules belonging to a sample. With the help of electric fields and magnetic
fields, the method of mass spectroscopy is used to measure the ratio of the mass of the
molecule to its charge. A common instrumental method used in the field of analytical
chemistry is electrochemical analysis. In this method, the analyte is placed in an
electrochemical cell and the voltage or the current flowing through it is measured.
The interaction between the analyte and energy in the form of heat is studied in the
discipline of analytical chemistry known as calorimetry. A calorimeter is an instrument that
is used to measure the heat of a chemical reaction. Example:- Conductometric titrations, pH
metric titrations, Potentiometric titrations, Spectroscopic methods, Thermal Methods,
Electro analytical methods, etc.,.
The difference between qualitative and quantitative analysis in chemistry is that the
qualitative analysis does not measure the amount of the substance but measures the quality of
that material whereas quantitative analysis in chemistry gives the absolute or relative quantity
regarding the concentration of one or more substances present in a sample or compound
TYPES OF INSTRUMENTAL METHODS
1.. SPECTRO CHEMICAL METHOD – It Is the study of the interaction between matter and
radiated energy. Spectroscopy concept is now expanded to comprise any interaction with
radiative energy as a function of its wavelength or frequency. Spectroscopic data is often
represented by a spectrum, a plot of the response of interest as a function of wavelength or
frequency. Examples: -Atomic absorption spectroscopy -Flame emission spectroscopy
-Visible and ultraviolet spectroscopy -Infrared spectroscopy -Nuclear magnetic resonance
spectroscopy.
2.. ELECTRO CHEMICAL METHOD- It Is the study of chemical reactions which take place at
the interface of an electrode, usually a solid metal or a semiconductor, and an ionic
conductor, the electrolyte.
These reactions involve electric charges moving between the electrodes and the electrolyte
(or ionic species in a solution). Thus electrochemistry deals with the interaction between
electrical energy and chemical change, and involve measurement of potential, current, or
charge in an electrochemical cell that serves as the analytical signal. Examples: -
Potentiometric methods -Coulometric methods -Voltammetric methods
3.. THERMAL METHOD – It Is a branch of materials science where the properties of
materials are studied as they change with temperature. Several methods are commonly
used, these are distinguished from one another by the property which is measure.
Examples: -Dielectric thermal analysis (DEA) Measure dielectric permittivity and loss factor
, Differential thermal analysis (DTA) Measure temperature difference , Differential scanning
calorimetry (DSC) Measure heat difference, Dilatometry (DIL) Measure volume,
Thermogravimetric analysis (TGA) Measure mass.
4.. SEPARATION METHOD – It Is a method to achieve any mass transfer phenomenon that
converts a mixture of substances into two or more distinct product mixtures or its pure
constituents. Separations are carried out based on differences in chemical properties, or
physical properties such as size, shape, mass, density, boiling and melting points, solubility,
or chemical affinity, between the constituents of a mixture, and are often classified according
to the particular differences they use to achieve separation. Usually there is only physical
movement, no substantial chemical modification. Examples: -Solvent extraction,
Chromatography, Electrophoresis.
APPLICATION OF ANALYTICAL CHEMISTRY
1.. The shelf lives of many medicines are determined with the help of analytical chemistry.
2.. It is used to check for the presence of adulterants in drugs. 3.. Soil can be tested to check
for appropriate concentrations of minerals and nutrients that are necessary for plant
growth. 4.. It is employed in the process of chromatography where the blood samples of a
person are classified. 5.. The concentration of the pesticide residues and the contaminants
in a given food sample can also be determined via analytical chemistry. 6.. It also has many
important applications in medicine, with its use in the testing of cholesterol and glucose
levels in a blood sample. 7.. Analytical chemistry is an integral part of forensic science,
clinical analysis, and even environmental analysis.
CLASSIFYING ANALYTICAL TECHNIQUES
Consider the two graduated cylinders in, each of which contains a solution of 0.1 M Cu(NO3)2.
Cylinder 1 contains 10 mL, or 1.0×10−4 moles of Cu2+, and cylinder 2 contains 20 mL, or
2.0×10−4 moles of Cu2+. If a technique responds to the absolute amount of analyte in the
sample, then the signal due to the analyte SA SA=kAnA -----------1
where nA is the moles or grams of analyte in the sample, and kA is a proportionality constant.
Because cylinder 2 contains twice as many moles of Cu2+ as cylinder 1, analyzing the contents
of cylinder 2 gives a signal twice as large as that for cylinder 1.
A second class of analytical techniques are those that respond to the analyte’s concentration,
CA SA=kACA ---------------2
Since the solutions in both cylinders have the same concentration of Cu2+, their analysis yields
identical signals.
A technique that responds to the absolute amount of analyte is a total analysis technique.
Mass and volume are the most common signals for a total analysis technique, and the
corresponding techniques are gravimetry and titrimetry. With a few exceptions, the signal for a
total analysis technique is the result of one or more chemical reactions, the stoichiometry of
which determines the value of kA in Equation 1.
Spectroscopy and electrochemistry, in which an optical or an electrical signal is proportional to
the relative amount of analyte in a sample, are examples of concentration techniques. The
relationship between the signal and the analyte’s concentration is a theoretical function that
depends on experimental conditions and the instrumentation used to measure the signal. For
this reason the value of kA in Equation 2 is determined experimentally.
Selecting an Analytical Method A method is the application of a technique to a specific
analyte in a specific matrix. We can develop an analytical method for determining the
concentration of lead in drinking water using any of the techniques mentioned in the previous
section. A gravimetric method, for example, might precipitate the lead as PbSO4 or PbCrO4, and
use the precipitate’s mass as the analytical signal. Lead forms several soluble complexes, which
we can use to design a complexation titrimetric method. we can use graphite furnace atomic
absorption spectroscopy to determine the concentration of lead in drinking water. Finally, the
availability of multiple oxidation states (Pb0 , Pb2+, Pb4+ ) makes electrochemical methods
feasible. The requirements of the analysis determine the best method. In choosing a method,
consideration is given to some or all the following design criteria: accuracy, precision,
sensitivity, selectivity, robustness, ruggedness, scale of operation, analysis time, availability of
equipment, and cost.
ACCURACY Accuracy is how closely the result of an experiment agrees with the “true” or
expected result. We can express accuracy as an absolute error, e
e = obtained result - expected result
or as a percentage relative error, %er
%er = ((obtained result − expected result) / expected result) × 100
Analytical methods may be divided into three groups based on the magnitude of their relative
errors.3 When an experimental result is within 1% of the correct result, the analytical method
is highly ac- curate. Methods resulting in relative errors between 1% and 5% are moderately
accurate, but methods of low accuracy produce rel- ative errors greater than 5%.
A method’s accuracy depends on many things, including the signal’s source, the value of kA in a
solution and the ease of handling samples without loss or contamination. In general, methods
relying on total analysis techniques , such as gravimetry and titrimetry, produce results of
higher accuracy because we can measure mass and volume with high accuracy.
PRECISION When a sample is analyzed several times, the individual results are rarely the
same. Instead, the results are randomly scattered. Precision is a measure of this variability. The
closer the agreement between individual analyses, the more precise the results.
a Searching for chemicals, reagents, or apparatus can waste your time and delay the reaction-
b the weight of an object of 100g to within ± 0.01 mg. analytical balances are frequently used in
For example, in determining the concentration of K+ in serum, the results shown in Figure a
are more precise than those in Figure b. It is important to realize that precision does not imply
accuracy. That the data in Figure a are more precise does not mean that the first set of results is
more accurate. In fact, both sets of results may be very inaccurate. As with accuracy, precision
depends on those factors affecting the relationship between the signal and the analyte (eqn 1
and 2). Of particular importance are the uncertainty in measuring the signal and the ease of
handling samples reproducibly. In most cases the signal for a total analysis method can be
measured with a higher precision than the corresponding signal for a concentration method.
SENSITIVITY The ability to demonstrate that two samples have different amounts of analyte is
an essential part of many analyses. A method’s sensitivity is a measure of its ability to establish
that such differences are significant. Sensitivity is often confused with a method’s detection
limit, which is the smallest amount of analyte that we can determine with confidence.
Sensitivity is equivalent to the proportionality constant, kA, in equation 1 and equation 2 If DSA
is the smallest difference that we can measure between two signals, then the smallest
detectable difference in the absolute amount or relative amount of analyte is
∆nA = ∆SA/kA or ∆CA = ∆SA/Ka
SPECIFICITY & SELECTIVITY An analytical method is specific if its signal depends only on the
analyte. 4 Although specificity is the ideal, few analytical methods are completely free from the
influence of interfering species. When an interferent contributes to the signal, we expand
equation 3.1 and equation 3.2 to include its contribution to the sample’s signal, Ssamp
Ssamp = SA + SI = kAnA + kInI  Ssamp = SA + SI = kACA + Kici
where SI is the interferent’s contribution to the signal, kI is the interferent’s sensitivity, and nI
and CI are the moles (or grams) and concentration of the interferent in the sample.
Selectivity is a measure of a method’s freedom from interferences. The selectivity of a method
for the interferent relative to the analyte is defined by a selectivity coefficient, KA,I
KA,I = kI / kA
which may be positive or negative depending on the sign of kI and kA. The selectivity
coefficient is greater than +1 or less than –1 when the method is more selective for the
interferent than for the analyte. Determining the selectivity coefficient’s value is easy if we
already know the values for kA and kI.
ROBUSTNESS & RIGIDNESS For a method to be useful it must provide reliable results.
Unfortunately, methods are subject to a variety of chemical and physical interferences that
contribute uncertainty to the analysis. When a method is relatively free from chemical
interferences, we can use it on many analytes in a wide variety of sample matrices. Such
methods are considered robust. Random variations in experimental conditions also introduces
uncertainty. If a method’s sensitivity, k, is too dependent on experimental conditions, such as
temperature, acidity, or reaction time, then a slight change in any of these conditions may give
a significantly different result. A rugged method is relatively insensitive to changes in
experimental conditions.
SCALE OF OPERATION Another way to narrow the choice of methods is to consider three
potential limitations: the amount of sample available for the analysis, the expected
concentration of analyte in the samples, and the minimum amount of analyte that will produce
a measurable signal. Collectively, these limitations define the analytical method’s scale of
operations.
NEATNESS & CLEANLINESS
The objectives of Neatness & Cleanliness include prevention of possible contamination and
cross-contamination Contamination by a variety of substances contaminants (e.g. microbes,
previous
products, residues of cleaning agents, airborne materials (e.g. dust and particulate matter),
lubricants and ancillary material, such as disinfectants Also decomposition residues from
product or detergents.
Importance of Cleanliness in Laboratories :- A laboratory is one of the potentially dangerous
places that should be taken care of adequately. The accuracy of the laboratory results is highly
proportional to the hygiene conditions of the laboratory. An untidy and cluttered workplace
can negatively influence the experimenter and add unnecessary stress.
A wide range of different glass, ceramic or plastic vessels are used in laboratories to perform
experiments and analyses, to culture tissue, take specimens and many other applications.
Ensuring that standards of apparatus cleanliness do not jeopardize the outcome of experiments
is of paramount importance in laboratories and requires complex treatment processes and
excellent efficiency. Good laboratory practice demands clean glassware, because the most
carefully executed piece of work may give a flawed result if dirty glassware is used. In all
instances glassware must be - physically clean, - it must be chemically clean, - must be
microbiologically clean or sterile.
Keeping the laboratory glassware physically neat and clean, free of grease, and bacteria is
therefore of the utmost importance.
Here are some reasons why you should prioritize the cleanliness of laboratories
Provide accurate results An unclean laboratory is as harmful as an inaccurate result. The
apparatus, especially the glassware, used in labs must be cleaned after every use. The remnants
of previous tests, moisture, or even dust particles can alter the laboratory results. This can
result in a waste of money and energy.
Avoid contamination To get highly reliable and accurate results, some test samples are stored
under optimal environmental conditions. This needs an environment that protects the samples
from cross-contamination from other samples and the absence of dust particles in the vicinity
of the samples. Using IPA to clean the surfaces will help to minimize cross-contamination as it
removes all kinds of contaminants in laboratories.
Comply with the safety regulations
The laboratories are potentially dangerous places governed by strict health and safety policies.
To comply with the health and safety policies, they should be kept clean and properly sanitized.
Ignoring these would lead to the cancellation of license or the permanent shut down of your
lab.
Stay organized
To arrange your pieces of equipment in the proper configuration, you have to organize your
laboratory to the optimal level. You must remove leftover debris and residue to keep the labs
clean and sanitized. An organized laboratory keeps everything within your reach and helps you
to conduct experiments at a smooth pace.
Improve the work efficiency
The overall efficiency of a laboratory test is affected by the cleanliness and hygiene of a
laboratory. A well-organized laboratory can make you motivated and save your time from
locating various bottles, apparatus, etc. It de-clutters the laboratory and helps you to glide from
one step to the other without confusion.
Prevention of breakages
Bottles and reagents kept haphazardly are prone to breakages when they are handled
carelessly. They should be arranged on their shelves with proper labels. Cluttering the debris
with bottles and glassware can lead to accidental spills, slips, rollovers, and breakages.
Saves money
Maintaining proper hygiene in your laboratory can curb extra expenditure on lab-equipments.
An untidy and unorganized workplace adds extra cost to the experiments. Accidental
breakages and spillages can waste money spent on lab apparatus and chemicals. Contaminants
can affect the accuracy of the results which would require repetition of the experiment.
process. It can also slow down your workflow and reduce the efficiency of your workplace.
ANALYTICAL BALANCE FULL
Analytical balances are an extremely accurate laboratory balance created to precisely
measure the mass of an object. Offering readability up to 0.00001 grams (0.01 mg), It detects
laboratories. Providing such accurate measurement means that the balance is highly
sensitive. Analytical balances are therefore designed with draft shields to provide protection
from external environments such as air flows and dust which might affect the precision.
Draw the DIAGRAM
Balance plate (Pan)- A container to bold the sample material for mass measurement.
Weights- It enables the calibration of analytical scales. Power button (on/off button)- It is
used to activate or deactivate the balance. ‘Re-zero’ or ‘Tare’ button- It is used to rebalance
the system and bring it back to neutral (zero) ‘Mode’ button- It is used to configure the
measurement conversion system such that the conversion system can be changed as necessary.
Draft shields- These are incorporated into the design of analytical balances to offer protection
from outside factors such as air flows and dust that could compromise precision.
Level adjustment feet- These enable the balance to be brought to the reference position.
These are movable legs. The leveling bubble, spirit level, or plumb bob determines the
reference position. Level indicator- It checks the balance in level. Display panel- It displays
various information such as results, errors, information for function settings, and function in
progress.
WEIGHING PROCEDURE
Select a proper location
Several external environmental factors influence the measurement. Hence, it’s necessary to
carry out the weighing procedures in a suitable location.
Choose a stable and horizontal location free from external disturbances., Avoid direct sunlight
and make sure there are no extreme temperature changes., Refrain from touching magnetic or
magnetic field-generating objects or equipment., The environment should be as dust-free as
possible., Steer clear of air currents produced by ventilators, air conditioners, open doors, and
windows.
Leveling the analytical balance
Repeatable measurements and precise findings require precise horizontal positioning., The
analytical balance must be leveled to account for any slight deviations or tilts at this site. Until
the air bubble in the indicator is in the center, the analytical balance’s leveling feet should be
adjusted.
Calibrating the analytical balance
The analytical balance must be calibrated for the sample to be precisely weighed. The following
circumstances call for calibrating the operations: Modifications to the usage location (including
moving within the same room)., Alteration in the environment., Before each use In-built
internal calibration or external calibration can be performed. Internal calibration requires
manual input from the users to reset the analytical balance, and external calibration is
performed using a calibration pendulum.
Weighing
It is preferable to preheat it for an hour before using it., Set the analytical balance to zero in the
no-load condition by pressing the “tare” button., Place the weigh boat, weigh paper, or other
vessel or container in the center of the weighing pan and then shut the glass door of the
weighing chamber., Check the value that was displayed after it was stabilized. The appearance
of the stability mark indicates a stable state., To exclude container mass from the measurement,
the ‘TARE’ button is pressed to reset the mass to zero., Add the substance to be weighed after
removing the container from the balance. Avoid putting things in the balance pan because
doing so can contaminate the balance., Reset the container’s balance, then wait 5-10 seconds
(up to a minute) for the mass reading to stabilize.
Cleaning
The analytical balance’s measurement accuracy and lifespan are improved by routine cleaning.
Use just a piece of lint-free, soap-wet, mild detergent-coated cloth to clean the analytical
balance., Avoid using any abrasive or harsh cleaning chemicals as well as organic solvents.
Cut off the electricity and unplug the power cord while cleaning.
Ensure no liquid or dust gets inside the housing of the analytical balance.
ERRORS IN ANALYTICAL BALANCE
Placing a sensitive scale near heavy machinery can result in vibrations being picked up by the
load cell., Moving air can push a scale down creating false “weight”., Scale components that rub
aggressively against each other can cause tiny vibrations which are picked up by the load
cell(Friction). Load cell values will stray from their true reading over time(Lack of calibration)
WEIGHING TECHNIQUES
A laboratory balance is used to determine the weight of an object. There are at least three types
of balance used in the laboratory name Triple Beam Balance, Top Loading Balance and
Analytical Balance. all of which differ in design and in their level of accuracy. There are two
types of weighing techniques that comes in handy when performing laboratory experiments
namely: (1) Direct Weighing and the (2) Indirect Weighing or Weighing by difference.
Direct Weighing
1. Using a 4”x 5” wax paper / petri dish, create three pieces of origami box.
2. Place the box paper / petri dish in the triple beam balance and re- zero/tare the balance.
3. Place a coin, prepared by the instructor, on the paper box. 4. Obtain the weight of the coin
by reading the calibration of the balance and record the data on the Data Sheet. 5. Repeat steps
1 - 5 but this time using the Top Loading Balance and Analytical Balance using the same coin.
Indirect Weighing or Weighing by Difference
1. Create three origami box. 2. Place the paper / petri dish in the triple beam balance and
record the weight of the paper and label as Wt. of Paper box / Wt. of Petri dish.
3. Place a coin on the paper box. 4. Obtain the weight of the coin by reading the calibration,
label this data as Wt. of Paper box / Wt. of Petri dish + Wt. of Coin.
5. Record the data on the Data sheet. The weight of the coin is the difference between the Wt. of
Paper box / Wt. of Petri dish + Wt. of Coin and Wt. of Paper box. 6. Repeat steps 1 - 5 but this
time using the Top Loading Balance and Analytical Balance using the same coin.
VOLUMETRIC GLASSWARE
In Analytical chemistry(quantitative), it is often necessary to make volume measurements
with an error on the order of 0.1%, one part per thousand. This involves using glassware that
can contain or deliver a volume known to a few hundredths of a milliliter, or about 0.01 mL.
One can then report quantities greater than 10 mL to four significant figures. Glassware
designed for this level of accuracy and precision is expensive, and requires some care and skill
to give best results. Four main types of volumetric glassware are common: the graduated
cylinder, the volumetric flask, the buret and the pipet. These have specific uses and will be
discussed individually. There are some points that are common to all types, however. These
involve cleanliness and how to read volumes accurately.
All volumetric glassware is calibrated with markings used to determine a specific volume of
liquid to varying degrees of accuracy. To read this volume exactly, the bottom of the curved
surface of the liquid, the meniscus, should be located at the scribed line for the desired volume.
It is often easier to see the meniscus if you put a white paper or card behind the apparatus. If
your eye is above or below the level of the meniscus, your readings will be inaccurate due to
the phenomenon of parallax. View the meniscus at a level perpendicular to your eye to avoid
this as a source of error.
1.. Graduated Cylinder - Most students are familiar with graduated cylinders, which are used
to measure and dispense known volumes of liquids. They
are manufactured to contain the measured volume with
an error of 0.5 to 1%. For a 100 mL graduated cylinder,
this would be an error of 0.5 to 1.0 mL. Measurements
made with a graduated cylinder can be reported to three
significant figures.
2.. VOLUMETRIC FLASK - The volumetric flask, available in sizes ranging from 1 mL to 2 L,
is designed to contain a specific volume of liquid, usually to
a tolerance of a few hundredths of a milliliter, about 0.1%
of the flask's capacity.
A volumetric flask (measuring flask or graduated flask) is a
piece of laboratory apparatus, a type of laboratory flask,
calibrated to contain a precise volume at a certain
temperature.
Volumetric flasks are used for precise dilutions and preparation of standard solutions. These
flasks are usually pear-shaped, with a flat bottom, and made of glass or plastic. The flask's
mouth is either furnished with a plastic snap/screw cap or fitted with a joint to accommodate
a Poly Tetra Fluoro Ethylene (PTFE) or glass stopper. The neck of volumetric flasks is
elongated and narrow with an etched ring graduation marking. The marking indicates the
volume of liquid contained when filled up to that point. The marking is typically calibrated "to
contain at 20 °C and indicated correspondingly on a label. The flask's label also indicates the
nominal volume, tolerance, precision class, relevant manufacturing standard and the
manufacturer's logo. Volumetric flasks are of various sizes, containing from a fraction of a
milliliter to hundreds of liters of liquid.
CALIBRATION OF VOLUMETRIC FLASK - The flask must be completely dry. Using the top
loading balance, weigh the dry empty flask with its stopper and measure the mass, recording
all the decimal places. Fill the flask exactly to the mark with distilled water that has been
allowed to reach room temperature. Any drops of water clinging to the inside of the neck of
the flask must be removed with a rolled-up piece of paper towel, so try to avoid this. Weigh the
flask, stopper, and water. Calculate the true volume of the flask using the conversion data in
Table 1-1. Repeat the determination empty the flask and refill) until you are confident that
you have the correct volume with an uncertainty of less than one part per thousand.
Volumetric flasks must NEVER be heated, either with an open flame or on a hot plate. The high
temperature causes irreversible changes in the shape of the glass and the flask must be
recalibrated. Later in the term, DI water is heated up and for the same reason; it needs to be
cooled down to room temperature before use in order to obtain the calibrated volume.
CLEANING OF VOLUMETRIC FLASK – Volumetric flasks can be cleaned using the detergent
washing and distilled deionized water rinsing procedures described above. The inside of the
flasks can be soaked by putting 1-2 pumps of working solution in first with the pump bottle
next to the sink in Room 203 before filling with tap water. Follow soaking times as if you were
soaking the flask inside a Citronox bath. Vigorous shaking of the detergent solution should aid
in cleaning the inside of the flask. Acid rinses may be used if the solutions prepared are likely
to resist detergent cleaning. Do not allow alkaline solutions to be stored for long periods of
time in volumetric flasks as they may become damaged. After cleaning a volumetric flask, fill
with deionized water before returning it to storage. Discard water before using flask again.
3.. BURET- A buret is a long, narrow tube with a stopcock at its base. It is used for accurately
dispensing variable volumes of liquids or solutions. It is graduated in 0.1 mL increments, with
the 0.00 mL mark at the top and the 50.00 mL mark near the bottom. Notice that the marks do
not go all the way to the stopcock. Therefore the buret actually will hold more than 50.00 mL
of solution. Burets with liquid capacities of 25.00 mL and 10.00 mL are also available.
CLEANING OF BURET - For optimal accuracy and to prevent contamination, a buret must be
clean. To test a buret for cleanliness, close its stopcock and pour a small volume (5-10 mL) of
deionized water into it. Hold the buret at a slant, almost parallel to the desk surface. Slowly
rotate the buret and allow the liquid to coat its inside surface. Then hold it upright; the liquid
should settle to the bottom of the buret in sheets, leaving no droplets on the interior walls. If
droplets form on the walls, wash the inside with a soap solution, and rinse with distilled or
deionized water. Repeat the cleanliness test. Just before use, a buret should be "conditioned"
to ensure that any water adhering to the inside walls is removed. Add ~5 mL of the liquid that
is to be used into the buret. Rinse the walls of the buret, then drain the liquid through the
stopcock. Repeat with a second volume of liquid
CALIBRATION OF BURET - For calibration purposes of this equipment, these steps can be
followed: Take a clean conical flask of 100cm3 fitted with a cork. Measure the mass of the flask
accurately and record it to the nearest milligram on the analytical balance Mass. Clean the
burette thoroughly and grease the stopcock properly. Fill up the burette with distilled water
and note the room temperature. Make sure that the water level must be leveled to the zero
mark. Touch the burette tip to the wall of the beaker to remove the adhering liquid drop. Take
the initial reading of the water level in the burette. Pour 10 cm2 of distilled water into the
conical flask. Touch the tip of the burette to the wall of the flask. Cork the flask and weigh it
accurately on an analytical balance and record the mass. The difference between this mass and
the initial mass of the flask gives the mass of the water dispensed. calculate the amount of
Water is actually dispensed by dividing the mass by the density of the water. Repeat the same
process with 20, 30, 40 and 50 cm3 water, thus calibrating the burette over its entire length.
4. PIPET - A volumetric pipette, bulb pipette, or belly pipette have a large bulb with a long
narrow portion above with a single graduation mark as it is calibrated for a single volume (like
a volumetric flask). Typical volumes are 1, 2, 5, 10, 20, 25, 50 and 100 mL. Volumetric pipettes
are commonly used in analytical chemistry to make laboratory solutions from a base stock as
well as to prepare solutions for titration. the standard tolerance for volumetric transfer
pipettes depends on the size: a 0.5-mL pipette has a tolerance of ±0.006 mL, while a 50-mL
pipette has a tolerance of ±0.05 mL. A specialized example of a volumetric pipette is the micro
fluid pipette (capable of dispensing as little as 10 µL) designed with a circulating liquid tip that
generates a self-confining volume in front of its outlet channels.
CLEANING OF PIPET – In order for the pipet to deliver the proper amount of solution, it must
be properly cleaned. Using the appropriate technique for filling the pipet, draw distilled water
into the pipet until it almost reaches the bulb. Remove the bulb and allow the water to drain. If
the water drains without leaving droplets on the inside of the pipet, repeat this rinse twice
more and go on to step two. If droplets cling to the inside of the pipet, clean it with detergent
solution or cleaning solution. Rinse many times with tap water, then repeat the distilled water
rinses., Draw a small amount of the solution you wish to pipet into the pipet. Tilt and roll the
pipet so that the solution has contact with all the inside surfaces. Discard the solution from the
rinses. Repeat this rinse at least once more., When you are finished with the pipet in your
experiment, rinse with distilled water.
CALIBRATION OF PIPET - Take a clean 100 cm3 conical flask fitted with a cork or stopper.
Weigh the conical flask with cork accurately on an analytical balance to the nearest milligram
and record its mass in your notebook. Pipette out 10/20/25 cm3 (equal to the capacity of the
pipette) of distilled water from a beaker into the conical flask and weigh the flask with its
contents. Record the temperature of the water with a thermometer. Calculate the mass of
water delivered from the
difference of the two readings. Repeat the procedure 3-4 times. Take average mass and divide
it by density of water at room temperature to calculate the actual capacity of pipette.
GENERAL CLEANING OF VOLUMETRIC GLASSWARE It's generally easier to clean glassware if
you do it right away. When detergent is used, it's usually one designed for lab glassware, such
as Liquinox or Alconox. These detergents are preferable to any dishwashing detergent you
might use on dishes at home. Much of the time, detergent and tap water are neither required
nor desirable. You can rinse the glassware with the proper solvent, then finish up with a
couple of rinses with distilled water, followed by final rinses with deionized water.
Volumetric Glassware should be washed in hot water or put through a glassware washer as
soon as it is no longer in use to prevent the accumulation of residue that is difficult to remove
if it is not feasible to clean glassware immediately after use., There are a variety of soaps,
detergents, and cleaning powders that may be used while washing laboratory glassware.
Cleaning solutions that have a slight abrasive effect will yield better results on particularly
unclean glassware. However, it is important to ensure that the abrasive does not harm the
glass., Attempt a lengthy soak in a mild solvent if basic cleansers don’t get the job done, or if
there is material in areas where brushes can’t reach. Both of these scenarios are possibilities.
If you soak the substance in hot water or shake it gently, you can make it work better., When
cleaning particularly unclean volumetric glassware, it is often essential to use cleaning
solutions with a stronger chemical component. These solutions are often quite corrosive; they
may include strong acids or bases, and they have the potential to inflict harm. Some types of
volumetric glassware must be sterilized prior to the cleaning process. This includes glassware
that has blood clots on it as well as glassware that has viruses or bacteria that produce spores
on it. Glassware may be sterilized in autoclaves, steam ovens, or by boiling it for thirty minutes
in water that has between one and two percent of mild detergent or soft soap added to it.
The diagonal lines connecting the axes show combinations of sample size and analyte
concentration that contain the same absolute mass of analyte. As shown in Figure 3.4.2 , for
example, a 1-g sample that is 1% w/w analyte has the same amount of analyte (10 mg) as a
100mg sample that is 10% w/w analyte, or a 10-mg sample that is 100% w/w analyte If a
method’s minimum detectable signal is equivalent to 10 mg of analyte, then it is best suited to a
major analyte in a macro or meso sample. Extending the method to an analyte with a
concentration of 0.1% w/w requires a sample of 10 g, which rarely is practical due to the
complications of carrying such a large amount of material through the analysis. On the other
hand, a small sample that contains a trace amount of analyte places significant restrictions on an
analysis. For example, a 1-mg sample that is 10–4% w/w in analyte contains just 1 ng of analyte. If
we isolate the analyte in 1 mL of solution, then we need an analytical method that reliably can
detect it at a concentration of 1 ng/mL.
EQUIPMENT, TIME & COST Finally, we can compare analytical methods with respect to their
equipment needs, the time needed to complete an analysis, and the cost per sample. Methods that
rely on instrumentation are equipment-intensive and may require significant operator training.
For example, the graphite furnace atomic absorption spectroscopic method for determining lead
in water requires a significant capital investment in the instrument and an experienced operator
to obtain reliable results. Other methods, such as titrimetry, require less expensive equipment
and less training.
The time to complete an analysis for one sample often is fairly similar from method-to-method.
This is somewhat misleading, however, because much of this time is spent preparing samples,
preparing reagents, and gathering together equipment. Once the samples, reagents, and
equipment are in place, the sampling rate may differ substantially. For example, it takes just a few
minutes to analyze a single sample for lead using graphite furnace atomic absorption
spectroscopy, but several hours to analyze the same sample using gravimetry. This is a significant
factor in selecting a method for a laboratory that handles a high volume of samples.
The cost of an analysis depends on many factors, including the cost of equipment and reagents,
the cost of hiring analysts, and the number of samples that can be processed per hour. In general,
methods that rely on instruments cost more per sample then other methods.
MAKING THE FINAL CHOICE
Unfortunately, the design criteria discussed in this section are not mutually independent
[Valcá rcel, M.; Ríos, A. Anal. Chem. 1993, 65, 781A–787A]. Working with smaller samples or
improving selectivity often comes at the expense of precision. Minimizing cost and analysis time
may decrease accuracy. Selecting a method requires carefully balancing the various design
criteria. Usually, the most important design criterion is accuracy, and the best method is the one
that gives the most accurate result. When the need for a result is urgent, as is often the case in
clinical labs, analysis time may become the critical factor.
In some cases it is the sample’s properties that determine the best method. A sample with a
complex matrix, for example, may require a method with excellent selectivity to avoid
interferences. Samples in which the analyte is present at a trace or ultratrace concentration
usually require a concentration method. If the quantity of sample is limited, then the method
must not require a large amount of sample.
Determining the concentration of lead in drinking water requires a method that can detect lead at
the parts per billion concentration level. Selectivity is important because other metal ions are
present at significantly higher concentrations. A method that uses graphite furnace atomic
absorption spectroscopy is a common choice for determining lead in drinking water because it
meets these specifications. The same method is also useful for determining lead in blood where
its ability to detect low concentrations of lead using a few microliters of sample is an important
consideration.
GRAVIMETRIC ANALYSIS
Gravimetric analysis is a method of a quantitative assessment of laboratory techniques based
mostly on the dimension of an analyte's mass. One example of a gravimetric evaluation technique
may be used to decide the quantity of an ion in an answer by way of dissolving a regarded
quantity of a compound containing the ion in a solvent to break up the ion from its compound.
The ion is then induced or evaporated out of the answer and weighed. This form of gravimetric
evaluation is referred to as precipitation gravimetry. In this technique, compounds in an
aggregate are separated by way of heating them to chemically decompose the specimen. unstable
compounds are vaporized and misplaced (or gathered), leading to a measurable reduction within
the mass of the solid or liquid pattern.
The principle of gravimetric analysis is based on the estimation of the mass percent of an ion
in an impure compound of known quantity by determining the mass of the same ion in a pure
compound. In order to determine the mass, the ion of interest needs to be completely isolated.
This isolation of ions is done with the help of precipitation. Typical procedures used in
gravimetric analysis include:
Making a solution with a known amount of the
sample analyte., Separation of the desired
ion/element/radical in pure forms by various
separation methods., After the ion has been
separated, weigh the amount of the pure insoluble
compound formed., Calculating the value of the
individual component of interest, based on the
weight of the compound observed.
Gravimetric factor: A gravimetric factor is an
algebraic expression that converts grams of a
compound into grams of a single element. It is the
ratio of the formula weight (FW) of the substance
being sought to that of the substance weighed.
Ex - Fe is sought, Fe2O3 is weighed:
Types of Gravimetric Analysis There are 4 essential kinds of gravimetric analysis. Of which,
there are 2 common kinds regarding modifications within the section of the analyte to separate it
from the rest of a combination, resulting in an exchange in mass.
Volatilization Gravimetry Volatilization Gravimetry includes isolating components of our
mixture by means of heating or chemically decomposing the sample.
Volatilization methods can be either direct or indirect. Water eliminated in a quantitative manner
from many inorganic substances by ignition is an example of a direct determination. It is
collected on a solid desiccant and its mass determined by the gain in mass of the desiccant.
Another direct volatilization method involves carbonates which generally decompose to release
carbon dioxide when acids are used. Because carbon dioxide is easily evolved when heat is
applied, its mass is directly established by the measured increase in the mass of the absorbent
solid used
Precipitation Gravimetry Precipitation Gravimetry uses a precipitation reaction to split one or
more parts of a solution by incorporating it right into a strong solution.
In precipitation gravimetry an insoluble compound forms when we add a precipitating reagent,
or precipitant, to a solution containing our analyte. In most methods the precipitate is the
product of a simple metathesis reaction between the analyte and the precipitant; however, any
reaction generating a precipitate can potentially serve as a gravimetric method
Electrogravimetry It is a method used to separate and quantify ions of a substance, usually a
metal. In this process, the analyte solution is electrolyzed. Electrochemical reduction causes the
analyte to be deposited on the cathode. The mass of the cathode is determined before and after
the experiment, and the difference is used to calculate the mass of analyte in the original solution.
Controlling the potential of the electrode is important to ensure that only the metal being
analyzed will be deposited on the electrode.
Thermogravimetric Thermogravimetric is a technique of thermal analysis in which changes in
physical and chemical properties of substances are measured as a function of increasing
temperature or as a characteristic of time.
STEPS OF GRAVIMATRIC ANALYSIS
PREPARATION OF THE SOLUTION: This may involve several steps including adjustment of the
pH of the solution in order for the precipitate to occur quantitatively and get a precipitate of
desired properties, removing interferences, adjusting the volume of the sample to suit the
amount of ppt agent to be added.
PRECIPITATION: This requires addition of a precipitating agent solution to the sample solution.
Upon addition of the first drops of the precipitating agent, supersaturation occurs, then nucleation
starts to occur where every few molecules of precipitate aggregate together forming a nucleous. At
this point, addition of extra precipitating agent will either form new nuclei or will build up on
existing nuclei to give a precipitate.
DIGESTION OF THE PRECIPITATE: The precipitate is left hot (below boiling) for 30 min to 1 hour
in order for the particles to be digested. Digestion involves dissolution of small particles and
Re ppt on larger ones resulting in particle growth and better ppt characteristics. This process is
called Ostwald ripening. An important advantage of digestion is observed for colloidal ppt where
large amounts of adsorbed ions cover the huge area of the ppt. Digestion forces the small
colloidal particles to agglomerate which decreases their surface area and thus adsorption.
WASHING AND FILTERING THE PRECIPITATE: It is crucial to wash the precipitate very well in
order to remove all adsorbed species which will add to weight of precipitate. One should be
careful nor to use too much water since part of the precipitate may be lost. Also, in case of
colloidal precipitates we should not use water as a washing solution since peptization
would occur. In such situations dilute nitric acid, ammonium nitrate, or dilute acetic acid may be
used. Usually, it is a good practice to check for the presence of precipitating agent in the filtrate of
the final washing solution. The presence of precipitating agent means that extra washing is
required. Filtration should be done in appropriate sized Goosh or ignition filter paper
DRYING AND IGNITION: The purpose of drying or ignition in a muffle furnace at temperatures
ranging from 600-1200 oC is to get a material with exactly known chemical structure so that the
amount of analyte can be accurately determined.
PRECIPITATION FROM HOMOGENEOUS SOLUTION:In order to make Q minimum we can, in
some situations, generate the ppt agent in the precipitation medium rather then adding it.
Heating of the solution generates hydroxide ions from the hydrolysis of urea. Hydroxide ions are
generated at all points in solution and thus there are no sites of concentration. We can also
adjust the rate of urea hydrolysis and thus control the hydroxide generation rate. This type of
procedure can be very advantageous in case of colloidal precipitates
SAMPLE PREPARATION
In analytical chemistry, sample preparation (working-up) refers to the ways in which a sample is
treated prior to its analyses. Preparation is a very important step in most analytical techniques,
because the techniques are often not responsive to the analyte in its in-situ form, or the results
are distorted by interfering species. Sample preparation may involve dissolution, extraction,
reaction with some chemical species, pulverizing, treatment with a chelating agent (e.g. EDTA),
masking, filtering, dilution, sub-sampling or many other techniques. Treatment is done to prepare
the sample into a form ready for analysis by specified analytical equipment. Sample preparation
could involve: crushing and dissolution, chemical digestion with acid or alkali, sample extraction,
sample clean up and sample pre-concentration.
Sampling Methods
GASEOUS SAMPLING, LIQUID SAMPLING & SOLID SAMPLING
1.. Gaseous Sampling – It can be divided into two main categories which are Grab sampling &
Continuous or Integrated Sampling, in Grab sampling An actual sample of air is taken in a flask,
bottle or bag or other suitable container which is done over a period of few seconds or upto 1-2
minutes. While in Continuous sampling Gaseous or vapors are removed from the air over a
measured time period and concentrated by passage through a solid or liquid sorbent.
2.. Liquid Sampling - Liquid sample tend to be homogeneous and are easier to sample.
Liquids mix by diffusion only very slowly and must be shaken to obtain a homogeneous mixture.
If water sample is taken from the river, then the water samples is collected at the SURFACE,
MIDDLE and at the BOTTOM of the river bed. If the liquid is in a large container, then the liquid
should be stirred first before the samples are taken at the top, middle and at the bottom of the
container.
3.. Solid Sampling – In homogeneity of the solid sample, variation in sample size and variation
within the particle size make sampling of solids more difficult than other material. The easiest but
usually the most unreliable way to sample a solid material is by the grab sample, which is one
sample taken at random and assume to be representative. Solid samples usually need further
treatment.
There are some useful sampling techniques are below:-
Random Sampling The basis of random sampling is that each population unit has equal
probability of being selected. Random methods are good if the population does not have any
obvious trends or patterns.
Systematic sampling When a particular rule is fixed in sampling, for example if every 25th
sample is picked for analysis then the bais is to be followed
Systematic Random Sampling: The thumb rule of sampling or selecting a particular sample
is frequently and randomly changed then the sample is also called random systematic sampling.
Cluster sampling: If a bulk material is of different batches and in different sizes the same
sampling increment is collected from each batch (irrespective of its quantity in the bulk), the
sample is mixed together to get the gross sample. This sampling technique is called as cluster
sampling.
SAMPLE DISSOLUTION AND DECOMPOSITION
The objective of sample dissolution is to mix a solid or non-aqueous liquid sample quantitatively
with water or mineral acids to produce a homogeneous aqueous solution, so that subsequent
separation and analyses may be performed. Because very few natural or organic materials are
water-soluble, these materials routinely require the use of acids or fusion salts to bring them into
solution. These reagents typically achieve dissolution through an oxidation-reduction process that
leaves the constituent elements in a more soluble form.
There are some main techniques for sample decomposition discussed – fusion method, wet & Dry
ashing method, Dissolving method, Semi-melting method, microwave digestion.
Fusion Method Fusion method refers to mixing the sample with an acidic or alkaline solid
solvent, and allowing it to undergo a double decomposition reaction at high temperature, so
that the components to be measured are converted into water- or acid-soluble compounds,
such as sodium salts, potassium salts, sulfates or chloride, etc. Inorganic samples that are
insoluble in water, acid or alkali can generally be decomposed by this method.
Na2S2O7(403⦁C) or K2S2O7(419⦁C) heat Up to red heat with crucible (Pt, quartz, Porcelain)
gives decomposed samples of particularly those of Al, Be, Ta, Ti, Zr, Pu, and the rare earths.
Wet Ashing Method In this method, the mixture of nitric acid and sulfuric acid is usually
placed in a gram flask together with the sample, and digested at a certain temperature, in
which nitric acid can destroy most of the organic matter. During the digestion process, nitric
acid is evaporated, and finally sulfuric acid remains. When thick SO3 white smoke begins to
emerge, reflux is carried out in the flask until the solution becomes transparent. During
digestion, nitric acid oxidizes organic matter to carbon dioxide, water, and other volatile
products, leaving inorganic acids or salts. Ex- Hydrofluoride acid (Removal of silicon and
destruction of silicates; dissolves oxides of Nb, Ta, Ti, and Zr, and Nb, and Ta ores.)
Dry Ashing Method Dry Ashing is suitable for decomposing organic matter and biological
samples in order to determine the content of metal elements, sulfur and halogen elements. In
this method, the sample is heated and decomposed in a muffle furnace, and the oxygen in the
atmosphere acts as an oxidant, leaving inorganic residues after combustion. The residue is
usually leached with a small amount of concentrated hydrochloric acid or hot concentrated
nitric acid, and then quantitatively transferred to a glass container. In the dry ashing process, a
small amount of certain oxidizing substances (auxiliary agents) can be added to the sample as
needed to improve the ashing efficiency. Magnesium nitrate is one of the commonly used
additives. For liquid or wet animal and plant cell tissues, they should be dried by steam bath or
mild heating method before ashing and decomposition. The muffle furnace should be gradually
heated to the desired temperature to prevent ignition or foaming.
Dissolving Method Dissolution refers to the use of appropriate solvent to dissolve the sample
into a solution, this method is relatively simple and rapid. Water is one of the important
solvents to dissolve inorganic substances. Alkali metal salts, ammonium and magnesium salts,
inorganic nitrates and most alkaline earth metal salts are easily soluble in water. Acids, bases
or mixed acids are usually used as solvents for the decomposition of inorganic substances
insoluble in water.
Semi-melting Method The semi-melting method, also known as the sintering method, reacts
the sample with the flux at a temperature lower than the melting point. Compared with the
melting method, the semi-melting method has a lower temperature and a longer heating time,
but it is not easy to damage the dry pot, and it can usually be carried out in a porcelain crucible
without precious metal utensils.
Microwave digestion In addition to dissolving at room temperature and heating conditions,
microwave heating can also be used to assist dissolution. Microwave digestion uses the sample
and appropriate solvent (flux) to absorb microwave to heat the sample. At the same time, the
alternating magnetic field produced by microwave polarizes the dielectric molecules. The
alternating arrangement of polarized molecules in the high-frequency magnetic field leads to the
high-speed oscillation of the molecules, which makes the molecules obtain high energy. As a result
of these two actions, the surface layer of the sample is constantly agitated and broken, resulting in
rapid dissolution (melting). There are various microwave instruments that may be satisfactory
depending on sample preparation considerations. The three main approaches to microwave
dissolution are: focused open-vessel, low-pressure closed-vessel, and high-pressure closed-vessel.
Each has certain advantages and disadvantages and the choice of system depends upon the
application.
ADVANTAGE & DISADVANTAGE OF GRAVIMETRY ANALYSIS
ADVANTAGE - If the methods are followed carefully, it provides exceedingly precise analysis. It
is used to determine the atomic masses of many elements to six-figure accuracy. It provides
little room for instrumental error and does not require a series of standards for calculation of
an unknown.
DISADVANTAGE - It usually provides only for the analysis of a single element, or a limited
group of elements, at a time. Comparing modern dynamic flash combustion coupled with gas
chromatography with traditional combustion analysis.
THE LABORATORY NOTEBOOK
A laboratory notebook is most important tool when working in the lab. If kept properly, we
should be able to look back at our laboratory notebook several years from now and reconstruct
the experiments on which we worked.
laboratory data must be recorded directly into a bound, quadrille laboratory notebook that has
duplicate ("carbon-copy") pages. Reserve the first few pages for a table of contents, which
should be kept up-to-date. Entries should be legible and clear and made when an operation is
performed or when the data is generated. These entries should be made in lab notebook and
not on a loose piece of paper to be copied later. All entries must be in black or blue ink in the
English/Hindi language. Each page should have the day’s date at the top. If entries are made at
a later date, a new page should be used. Always include units of measurement when recording
data. Do not include any data that is not relevant to the lab. Errors should be crossed out with a
single line, not made illegible, just in case the "errors" later turn out to be important, and the
correction should be written near (above, below, or to the side if possible) the original entry.
The original information should still be able to be read. In industrial laboratories, it is required
that these corrections have a written explanation of why the correction was needed (typically
as a footnote). Most data should be handwritten, but it is permissible to tape a page into your
notebook if the 4 corners are marked so that it will be obvious if the page is removed. You
should also initial and date across one side of the page so that the partial initials and date are
visible on both the taped page and the notebook page..
SELECTING AND HANDLING OF REAGENTS
The purity of reagents has an important bearing on the accuracy attained in any analysis. It is,
therefore essential that the quality of reagents be consistent with their intended use and the
selection and handling of reagents and chemicals, and the Rules of Handling reagents and
Solutions. In order to get good results from analysis, the analyst must used suitable reagent.
If an analyst fails to use the correct grade of reagent, then the results obtained may not be
reliable. There are different grades of reagent for different purposes.
Reagent Grade Reagents-grade chemicals conform to the minimum standards set forth by the
reagents Chemical Committee of the American Chemical Society (ACS) and are used whenever
possible in analytical work. Some label their products with the maximum limits of impurity
allowed by the ACS specifications while others print actual concentrations for the various
impurities.
Primary Standard Grade Ultrapure compound that is used as reference in all volumetric
methods of analysis, Required for accurate volumetric analysis especially for preparing
standard solutions and reference standards. Primary standards reagents have been carefully
analyzed by the supplier, and the results are printed on the container label. The agency also
prepares and sells reference standards which are complex substances that have been
exhaustively analyzed.
Special-Purpose Reagents Chemicals Chemicals That have been prepared for a specific
application are also available. Included among these are solvents for spectrophotometric and
high-performance liquid chromatography. Information pertinent to the intended use is
supplied with these reagents. Data Provided with a spectrophotometric solvent, for example,
might include its absorbance at selected wavelengths and its ultraviolet cutoff wavelength.
Technical & laboratory grade Of lowest purity Suitable for non-critical tasks in the
laboratory such as rinsing, dissolving or are used as raw materials in production tasks.
Technical grade Do not have reference to establish purity. Other grades of reagent are also
available If correct reagent grade is used in analysis, then accuracy of results from the analysis
may be further improved.
Rules of Handling reagents and Solutions
A high-quality chemical analysis requires reagents and solutions of known purity. A freshly
opened bottle of a reagent-grade chemical can usually be used with confidence. Whether this
same confidence is justified when the bottle is half empty depends entirely on the way it has
been handled after being opened. We observe the following rules to prevent the accidental
contamination of reagents and solutions.
Choose the best grade of chemicals for analytical work. To weigh solid reagent, remove solid
by tapping the bottle gently on a table then pour the required quantity., Never insert spatulas,
spoon or knives into a reagent bottle., Always cap container immediately after removal of the
reagent., Always hold stoppers of reagent bottles between fingers; do not put the stopper on
the benchtop., Always clean the reagent shelf and the laboratory balance after use. Clean up
any spillages immediately., Always pour liquid reagent into a container. Never take the
reagent directly from its bottle by inserting pipette., Never return excess reagent back to its
bottle unless otherwise directed; you will contaminate the whole bottle by doing so.
PRECAUTIONS DURING SAMPLING
Before taking a sample must Rigorously sanitizing the hands with WHO-approved sanitizers
and rubbing the hands dry with sanitized and disposable hand towels.
After the sample has been collected, the technician will dispose off the used gloves after
leaving the sample container and wear a fresh pair for the next sample collection.
Thorough hand hygiene is observed before wearing gloves and after taking them off.
Technicians use one-time collection kits to ensure that new samples are not contaminated by
the remnants of the earlier samples.
After sampling, the sample containers must be checked for leaks. The outer surface of
packages must be clean and dry. If leaks occur, caps and stoppers should be reinforced or
replaced. Another inspection should then be carried out, and if leaks persist fresh samples
should be taken. Preferably use another sample container.
Warning signs, markings and symbols indicating potential hazards should be placed on
packages holding samples of hazardous goods/compounds.
Depending on your national regulations the sample container should be sealed in an
appropriate manner for the type of container used, to prevent unauthorised or inappropriate
handling of samples (and ensure the integrity of the contents). The seal must be firmly
attached and stable in order to prevent damage during sample storage or transport, and to
safeguard the chain of evidence.
The accompanying documents must be kept in line with rules laid down by the
administration. This depends on the situation. In some conditions only digital documents are
used; they are sent by email to the laboratory or using integrated information systems. Copies
of other relevant documents concerning the nature of the goods may also be enclosed.
The samples are to be placed in a sturdy leak-proof and water-proof bag/bottle/container.
The requisition form will go into a separate compartment.
THERMAL ANALYSIS 2.. T.G.A. (THERMO GRAVIMETRIC ANALYSIS) - Thermogravimetric analysis, or thermal
Thermal analysis is a branch of materials science where the properties of materials are studied as gravimetric analysis, (TGA) is a method of thermal analysis in which changes in physical and
they change with temperature. Properties are measured as a function of temperature, time, or chemical properties of materials are measured as a function of increasing temperature (with
Both Heat flow – direction, Mass change – loss / gain, Mechanical properties, Sheer, Strain, constant heating rate), or as a function of time (with constant temperature and/or constant
Dynamic loading, Gas evolution. mass loss). TGA can provide information about physical phenomena, such as second-order
Basic Principles of Thermal Analysis phase transitions, including vaporization, sublimation, absorption, adsorption, and desorption.
Modern instrumentation used for thermal analysis usually consists of the following parts: Likewise, TGA can provide information about chemical phenomena including chemisorptions,
▸ sample holder/compartment for the sample desolvation (especially dehydration), decomposition, and solid-gas reactions (e.g., oxidation or
‣ sensors to detect/measure a property of the sample and the temperature reduction).
‣ an enclosure within which the experimental parameters (temperature, speed, environment) TGA is commonly used to determine selected characteristics of materials that exhibit either
may be controlled ▸ a computer to control data collection and processing mass loss or gain due to decomposition, oxidation, or loss of volatiles (such as moisture).
Thermal analysis is a branch of materials science where the properties of materials are studied as Common applications of TGA are (1) materials characterization through analysis of
they change with temperature. Several methods are commonly used – these are distinguished characteristic decomposition patterns, (2) studies of degradation mechanisms and reaction
from one another by the property which is measured: kinetics, (3) determination of organic content in a sample, and (4) determination of inorganic
(e.g. ash) content in a sample, which may be useful for corroborating predicted material
1.. D.T.A. (DIFFRENTIAL THERMAL ANALYSIS) - structures or simply used as a chemical analysis. It is an especially useful technique for the
Differential thermal analysis (DTA) is a study of polymeric materials, including thermoplastics, thermosets, elastomers, composites,
thermoanalytic technique. similar to differential plastic films, fibers, coatings and paints.
scanning calorimetry. In DTA, the material under
study and an inert reference are made to undergo
identical thermal cycles, while recording any
temperature difference between sample and
reference. This differential temperature is then
plotted against time, or against temperature (DTA
curve, or thermogram). Changes in the sample,
either exothermic or endothermic, can be detected
relative to the inert reference. Thus, a DTA curve
provides data on the transformations that have
occurred, such as glass transitions, crystallization,
melting and sublimation. The area under a DTA
peak is the enthalpy change and is not affected by
the heat capacity of the sample.
A DTA curve can be used only as a finger print for identification purposes but usually the
applications of this method are the determination of phase diagrams, heat change measurements
and decomposition in various atmospheres. DTA is widely used in the pharmaceutical and food
industries.
3.. DSC or differential scanning calorimetry It is a thermoanalytical method that measures the
difference in the heat required to increase the temperature of a sample and the reference as a
function of temperature. In this method, both the sample and the reference are maintained at the
same temperature throughout the experiment.
Usually, the temperature used for the DSC method is designed as a temperature program in such
a way that the sample holder temperature increases linearly as a function of time. On the other
hand, the reference sample should have a well-defined heat capacity over the range of
temperatures that we are going to scan.
There are different types of DSC, such as heat-flux DSc and power differential DSC. Heat-reflux
DSC measures the difference in heat flux between the sample and a reference, whereas power
differential DSC measures the difference in power supplied to the sample and a reference.
INSTRUMENTATION OF DSC
Two basic types of DSC instruments: Heat flux DSC, Power compensated DSC
HEAT FLUX DSC
Sample Holder:- PLATINUM, ALUMINIUM, STAINLESS STEEL
SENSORS:- Temp Sensors, Usually thermocouples, which are same for the sample are reference.
Furnace:- One block for both sample and reference cells.
Temperature:- The temperature difference between the sample and reference is converted to
differential thermal power, which is supplied to the heaters to maintain the temperature of the
sample and reference at program value.
POWER COMPENSATED DSC
Sample Holder:- PLATINUM, ALUMINIUM,
STAINLESS STEEL PANS
SENSORS:- Pt resistance thermocouple, Separate
sensors & heaters for the sample & reference
sample.
Furnace:- Separate blocks for sample and
reference cells.
Temperature:- Differential thermal power is
supplied to the heaters to maintain the
temperature of the sample and reference at the
program level.
The key difference between TGA DTA and DSC is
that TGA measures the weight change of a sample
over a temperature range, while DTA measures
heat differences between a reference sample and
a sample of interest over a temperature range,
and DSC measures the heat flow of a sample over a temperature range.
DSC CURVE
The result of a DSC experiment is a curve of heat flux versus temperature or versus time. There
are two different conventions: exothermic reactions in the sample shown with a positive or
negative peak, depending on the kind of technology used in the experiment. •This curve can be
used to calculate enthalpies of transitions, which is done by integrating the peak corresponding
to a given transition. The enthalpy of transition can be expressed using equation: △H = KA
Where △H is the enthalpy of transition, K is the calorimetric constant, A is the area under the peak.
•The calorimetric constant varies from instrument to instrument, and can be determined by
analyzing a well-characterized material of known enthalpies of transition.
•Area under the peak is directly proportional to heat absorbed or evolved by the reaction,
•Height of the peak is directly proportional to rate of the reaction
Factors Affecting DSC Curve- There are two types of factors affecting DSC curve
1. Instrumental factors:
Instrumental factors: Furnace heating rate., Recording or chart speed, Furnace atmosphere,
Geometry of sample holder and location of sensors, Sensitivity of recording mechanism.,
Composition of sample container.
2. Sample Characteristics:
Amount of sample, Solubility of evolved gases in sample., Particle size, Heat of reaction, Sample
packing, Nature of sample, Thermal conductivity.
APPLICATION OF DSC
1 DSC Can be used to measure a number of characteristic properties of a sample., 2. using this
technique it is possible to observe fusion on crystallization events as well as glass temp.
transition temperature Tg 3.. DSC can also be used to study oxidation as well as other chemical
reactions. 4.. Can also be used to obtain valuable thermodynamic information about proteins.
5.. It is widely used for examing polymeric material to determine their thermal transitions . imp
thermal transitions include the glass transition temp.(Tg) , Crystalization temp. (Tc) & melting
temp.(Tm).
SAFETY IN ANALYTICAL LABORATORIES Standard Deviation MEAN
An Analytical Chemistry laboratory contains chemical products, solvents, acids, electricity, gas, A useful and commonly used measure of precision is the experimental standard The definition of mean is, "an average of n numbers computed by adding some function of the
glass equipment etc. and so is potentially dangerous. Being unaware of the properties of deviation defined by the VIM as... "for a series of n measurements of the same numbers and dividing by some function of n." The central tendency of a set of measurement
reagents and substances, not knowing how to use the equipment properly, or failing to measurand, the quantity s characterizing the dispersion of the results and given by the results is typically found by calculating the arithmetic mean (X bar ) and less commonly the
observe the safety norms in a laboratory can be a source of danger for you and everyone median or geometric mean. The mean is an estimate of the true value as long as there is no
formula:
present. Accidents can be avoided. In each case, the “ 3 Step Principle ” will be followed: systematic error. In the absence of systematic error, the mean approaches the true value (μ) as
Step 1: ELIMINATION OF DANGER (Search for alternatives)
s = [ ∑ (xi - x͡ )2 / (n-1) ]1/2
the number of measurements (n) increases. The frequency distribution of the measurements
Step 2: CONTAINMENT OF DANGER (Safety precautions) xi being the result of the i-th measurement and x͡ being the arithmetic mean of the n
approximates a bell-shaped curve that is symmetrical around the mean. The arithmetic mean is
Step 3: PROTECTION OF PEOPLE (Behavioral guidelines) results considered." calculated using the following equation:
There are some important procedure/steps for safety in a analytical lab which are mentioned The above definition is for estimating the standard deviation for n values of a sample of a = (X1 + X2 + ···Xn) / n
below population and is always calculated using n-1. The standard deviation of a population is Typically, insufficient data are collected to determine if the data are evenly distributed. Most
Eye Protection, Skin Protection, PPC, Hearing Protection, Respiratory Protection, First Aid. symbolized as s and is calculated using n. Unless the entire population is examined, s analysts rely upon quality control data obtained along with the sample data to indicate the
Eye Protection Mechanical effect, heat, chemicals or energy emissions can hurt the eyes. cannot be known and is estimated from samples randomly selected from it. For example, accuracy of the procedural execution, i.e., the absence of systematic error(s). The analysis of at
Experi-ence has shown that the handling of chemical materials, above all, and in more recent an analyst may make four measurements upon a given production lot of material least one QC sample with the unknown sample(s) is strongly recommended. Even when the QC
times, work with energy beams (lasers) can lead to eye injuries. In most cases, such injuries (population). The standard deviation of the set (n=4) of measurements would be sample is in control it is still important to inspect the data for outliers. There is a third type of
can be prevented by the use of suitable safety glasses . Safety glasses have relatively large, error typically referred to as a 'blunder'. This is an error that is made unintentionally. A
shatterproof lenses and an impact and heat resistant frame. When handling chemicals or
estimated using (n-1). If this analysis was repeated several times to produce several
blunder does not fall in the systematic or random error categories. It is a mistake that went
lasers, it is important that safety conscious behavior, above all, is observed. These glasses sample sets (four each) of data, it would be expected that each set of measurements unnoticed, such as a transcription error or a spilled solution. For limited data sets (n = 3 to 10),
protect your eyes better than the usual correction safety glasses since they provide side would have a different mean and a different estimate of the standard deviation. the range (Xn-X1), where Xn is the largest value and X1 is the smallest value, is a good estimate
protection in addi-tion to the safety lenses and frame already mentioned. If working with The experimental standard deviations of the mean for each set is calculated using the of the precision and a useful value in data inspection. In the situation where a limited data set
larger quantities of strongly corroding acids or caustic solutions, you should use closed eye following expression: s / (n)1/2 has a suspicious outlier and the QC sample is in control, the analyst should calculate the range
goggles and carry a face shield. You should not wear contact lenses when handling chemicals, Using the above example, where values of 1004, 1005, and 1001 were considered of the data and determine if it is significantly larger than would be expected based upon the QC
since they can strengthen the effect of chemicals penetrating between the lens and the eyes acceptable for the calculation of the mean and the experimental standard deviation the data. If an explanation cannot be found for an outlier (other than it appears too high or low),
and, in some cases, certain chemicals make the removal of the lenses diffi cult! When operating mean would be 1003, the experimental standard deviation would be 2 and the standard there is a convenient test that can be used for the rejection of possible outliers from limited
lasers, special laser eye protectors will be provided which provide protec-tion for the specifi c data sets. This is the Q test. The Q test is commonly conducted at the 90% confidence level but
deviation of the mean would be 1.
range of wavelengths of the laser. the following table includes the 96% and 99% levels as well for your convenience. At the 90%
Skin Protection Mechanical effects such as heat, cold, chemicals, microorganisms or energy confidence level, the analyst can reject a result with 90% confidence that an outlier is
beams can damage the skin. In a laboratory you can be exposed to many sources of danger. significantly different from the other results in the data set. The Q test involves dividing the
Skin protection, which consists at the very least of hand cream or gloves, must therefore be difference between the outlier and it's nearest value in the set by the range, which gives a
adapted to the respective sources of danger. Here, too, safety conscious behavior must fi rst be quotient - Q. The range is always calculated by including the outlier, which is automatically the
observed. One of the most frequent mechanical injuries, cuts to the hands, can be prevented by largest or smallest value in the data set. If the quotient is greater than the refection quotient,
the use of gloves while handling glass equipment and also by correct cutting techniques. When Q0.90, then the outlier can be rejected.
handling hazardous materials, thorough washing of the skin before and after the work in the The Q Test
laboratory is important.
Personal Protective Clothing Protective clothing, such as a laboratory coat, is important in n Q0.90 Q0.96 Q0.99
order to protect the whole body and clothes against the disturbing or harmful effects of dirt and 3 0.94 0.98 0.99
chemicals. In the case of contamination or fi re, it should be rapidly and easily removable. In 4 0.76 0.85 0.93
clean rooms, protective clothing is a necessity in order to prevent the contamination of the 5 0.64 0.73 0.82
rooms by agents such as road dust, etc. Lab coats are therefore mandatory in clean rooms. In 6 0.56 0.64 0.74
chemistry practical courses and work-shops, the use of a laboratory coat is either mandatory or 7 0.51 0.59 0.68
recommended. 8 0.47 0.64 0.53
Hearing Protection Ear protectors are sometimes necessary, particularly in workshops or 9 0.44 0.51 0.60
within the range of machine rooms. Remember that hearing damage is always irreversible.
10 0.41 0.48 0.57
There are various earphones or plugs, which provide protection for the hearing. It is
important that they are consistently worn, otherwise their protective effect is greatly reduced. Example: This example will test four results in a data set--1004, 1005, 1001, and 981.
For example, someone who works with a noise of 100 dB and wears no ear protection during The range is calculated: 1005 - 981 = 24. The difference between the questionable result
10% of his working time is still exposed to a (harmful) noise pollution of 90 dB. The Swiss (981) and its nearest neighbor is calculated: 1001 - 981 = 20. The quotient is calculated:
National Accident Insurance Fund (SUVA) has classifi ed a constant sound pressure level 20/24 = 0.83. The calculated quotient is compared to the Q0.90 value of 0.76 for n=4 (from
starting at 88 dB and lasting at least 8 hours per day as damaging to the hearing. table 14.3 above) and found to be greater. The questionable result (981) is rejected.
Respiratory Protection The respiratory system and lungs can be damaged by dust, chemical
gas or steam, and different types of gases. Inhalation of gas or steam can also be harmful to the
central or peripheral nervous system or to other organs such as the liver or kidney. Here, too,
safety conscious behavior is the cardinal rule for the prevention of accidents. For example,
chemical reactions which can develop harmful steams or gases should be performed under the
fume hood, with the ventilation switched on and the front shields closed whenever possible.
Find out, especially before starting a reaction, which substance could develop; some dan-
gerous gases such as carbon monoxide are odorless! With mechanical work you can, for
example, moisten the workpieces, in order to prevent the creation of dust. If this is not
possible, a ventilation source must be provided. The possibili-ties for respiratory protection
are easy enough, from a simple dust respirator over half masks to full masks with the
appropriate fi lter guards. If the oxygen supply is insuffi cient, this can be supplemented with
the use of compressed air breath-ing masks.
Fire Prevention Find out before beginning work at a laboratory about the locations and the
proper uses of the emergency showers and the fi re extinguishing equipment as well as the
escape routes. Limit the quantity of combustible liquids kept at the laboratory to a minimum
and keep them separate from strong oxidizing agents. For the storage of fl ammable liquids
that need to be stored cool, use refrigerators reserved only for this purpose and labeled.
Fume Hood All work in which poisonous, fl ammable or otherwise dangerous or foul -
smelling gas, steam, or aerosols could develop or escape should be performed under the fume
hood. The use of the fume hood is, nevertheless, not a carte blanche for the release of any
amount of substances and chemicals. Toxic and corrosive gas and steam, as mentioned in the
middle of the references written about devices, should be absorbed directly where they are
generated. This prevents disturbance of the envi-ronment and costly damage to the ventilation
system. Close the front sash of the fume hood whenever possible. Equipment should be placed
as far back as possible , against the back wall of the hood. Fume hoods equipped with switches
to regulate ventilation should, for environ-mental reasons, only be set at the highest setting
when in use. Otherwise, precious heat would escape unnecessarily.
Handling of Chemicals Beakers with chemicals must be covered and clearly labeled with at
least the following information:
Name of substance, Molecular formula, Date of filling, Name of responsible persons, Acids and
bases used for etching are highly corrosive. Before starting work, pay close attention to the fi
rst aid instructions for specifi c chemicals. Solvents may only be heated on the heating plate
under supervision. Do not clean hot heating plates with solvents. Use photo varnish only
under yellow light. Dispose of your chemicals after use, following the given guidelines
(introduction) and clean your glassware.
General lab safety rules
The following are rules that relate to almost every laboratory and should be included in most
safety policies. They cover what you should know in the event of an emergency, proper
signage, lab safety equipment, safely using laboratory equipment, and basic common-sense
rules.
Be sure to read all fire alarm and lab safety signs and follow the instructions in the event of an
accident or emergency. Ensure you are fully aware of your facility's/building's evacuation
procedures. Make sure you know where your lab's safety equipment—including first aid kit(s),
fire extinguishers, eye wash stations, and safety showers—is located and how to properly use
it. Know emergency phone numbers to use to call for help in case of an emergency. Lab areas
containing carcinogens, radioisotopes, biohazards, and lasers should be properly marked with
the appropriate warning signs. Open flames should never be used in the laboratory unless you
have permission from a qualified supervisor. Make sure you are aware of where your lab's
exits and fire alarms are located. An area of 36" diameter must be kept clear at all times around
all fire sprinkler heads. If there is a fire drill, be sure to turn off all electrical equipment and
close all containers. Always work in properly-ventilated areas. Do not chew gum, drink, eat, or
apply lip balm or cosmetics while working in the lab. Laboratory glassware should never be
used as food or beverage containers. Each time you use glassware, be sure to check it for chips
and cracks. Notify your lab supervisor of any damaged glassware so it can be properly disposed
of or recycled. Never use lab equipment that you are not approved or trained by your
supervisor to operate. If an instrument or piece of equipment fails during use, or isn't
operating properly, report the issue to a technician right away. Never try to repair an
equipment problem on your own. If you are the last person to leave the lab, make sure to lock
all the doors and turn off all ignition sources. Do not work alone in the lab. Never leave an
ongoing experiment unattended. Never lift any glassware, solutions, or other types of
apparatus above eye level. Never purposefully smell or taste chemicals. Do not pipette by
mouth. Make sure you always follow the proper lab safety procedures for disposing of lab
waste. Report all injuries, accidents, and broken equipment or glass right away, even if the
incident seems small or unimportant. If you have been injured, yell out immediately and as
loud as you can to ensure you get help. In the event of a chemical splashing into your eye(s) or
on your skin, immediately flush the affected area(s) with running water for at least 20 minutes.
If you notice any unsafe lab conditions, let your supervisor know as soon as possible.
ERROR
Errors in chemical analysis are simply defined as the difference between a measured value
and the true value. It denotes the estimated uncertainty in a measurement or experiment.
During chemical analysis, error in measurement occurs due to faulty calibration, standardization
or random variation, or uncertainty in results. By frequent calibration standardization and
analysis of the known sample, the error can be minimized but it is impossible to perform a
chemical analysis totally free of errors. In every chemical reaction, we want to minimize errors.
Types of Errors in chemical analysis
Errors are mainly of three types in chemical analysis:
Random error (Indeterminate error)
Systematic error (Determinate error)
Gross error
2. Indeterminate errors (Random error)
Data are distributed more or less symmetrically around the mean value as a result of the
indeterminate error. The precision of measurement reflects the random error. Hence,
measurement precision is impacted by random or indeterminate errors. Variable fluctuations
that are unavoidable or unknown that may have an impact on the findings of experiments are
what create indeterminate (or random) errors. Uncertainties in measurements can lead to
random or indeterminate errors. Errors of this kind, which are random or indeterminate, always
exist in measurements. Such an error can never be completely ruled out. They are a significant
factor in the determination of the analyte’s uncertainty. Most of the causes of random errors are
impossible to pinpoint. Due to the low value of individual causes of such errors, they cannot be
quantified even if their sources are known. However, the combined impact of all errors leads to
large variability in the measurement at random.
2. Determinate/Systematic error.
Determinate or systematic errors are those errors that have definite values and have some
assignable cause. For every repeated measurement carried out in the same manner, these errors
are consistently the same. Systematic errors usually introduce bias into the outcome of the
measurement. The accuracy of the results is influenced by significant mistakes. These mistakes
can also be identified and corrected because they are reproducible. Bias measures the
systematic error associated with analysis. It has a -ve sign if it causes results to be low and has
positive sign if the results are high.
Types of determinate error (systematic error)
Personal error: During the measurement of the analytical experiments, there is often a need for
personal judgment. For example: estimating the portion of the pointer between two scale
divisions, the color of the solution at the endpoint, the level of the liquid of the mark in pipette or
burette, etc. The main source of personal error is prejudice or bias. Human has a tendency to
estimate scale reading to the precision in a set of results. We sometimes knowingly cause the
result to fall closer to the true value. Number bias is also another important source of personal
error. Colour blindness person can cause a personal error in volumetric analysis.
2. Operative errors. These include personal errors and can be reduced by experience and care
of the analyst in the physical manipulations involved. Operative errors can be minimized by
having a checklist of operations. Operations in which these errors may occur include transfer of
solutions, effervescence and “bumping” during sample dissolution, incomplete drying of
samples, and so on. These are difficult to correct for. Other personal errors include mathematical
errors in calculations and prejudice in estimating measurements.
Instrumental or reagent error: The measuring tools also have a certain amount of
determinate error. Burette, pipette, and volumetric flask, for instance, always deliver slightly
differently from what the scale indicates. These inconsistencies primarily result from using the
glassware at a temperature that is different than the calibration since doing so damages the
container wall when it is heated to dry because of contamination on the interior surface. Due to
excessive use and the battery’s low voltage, electronic devices may also experience errors. The
failure to accurately and often calibrate the instruments could possibly be the cause of the
errors. Similar to how changing temperatures can affect numerous electronic components,
errors can result from these fluctuations.
Methodic error: The non-ideal type of chemical and physical behavior of reagents and reactions
on which an analysis is based can introduce methodic errors. The slowness of reaction, in the
completeness of reaction, un-stability of chemical species, and possible occurrence of side
reaction can cause methodic errors which may interfere with the measurement process. For
example, in volumetric analysis, a small amount of excess reagent is necessary to change the
color of an indicator to signify the completion of the reaction. The errors that are associated with
the methods are often very difficult to detect and the most serious type of error out of all types
of systematic error.
Constant or proportional error: Constant type of determinate errors are independent of the
size of the sample analyzed. When there are constant mistakes, the relative error changes as the
sample size is altered, but the absolute error remains constant. As the size of the quantity being
measured shrinks, the effect of constant error becomes more pronounced. Meanwhile,
Proportional errors decrease or increase in proportion to the size of the sample. The presence of
interfering impurities in the sample is the main cause of the proportional error. Iodine is
released, for instance, while estimating Cu++ with potassium iodide. If the samples are
contaminated with Fe+++ ions, I2 is also produced from KI, and an error in the estimation of Cu++
results. The amount of Fe+++ also doubles when the sample size is doubled, and error starts to
become compulsive. Such errors are referred to as additive errors since the proportional error is
sample size dependent.
3. Gross errors
Either too high or too low findings are the outcome of this kind of error. They are the outcome of
human mistakes. Outliers, or results that appear to differ significantly from all other measured
data in a set of repeated measurements, are frequently the result of gross error.
MINIMIZATION OF DETERMINATE ERROR
Indeterminate errors are beyond the control of the analyst, however, determinate errors can be
minimized. Some of the common methods that can be employed for the minimization of errors
are:
Calibration of apparatus: The instrument’s calibration is one method of reducing error.
Instruments should always be regularly calibrated because they could alter as a result of water,
corrosion, overuse, etc. The calibration can be done using (i) comparison with a standard, and
(ii) external standard calibration.
Blank determination: A determination is performed under identical conditions by excluding
the sample in order to reduce errors brought on by reagent impurities.
Independent method of analysis: This approach enables the parallel use of the technique that
is being evaluated and a reliable analytical strategy. The independent approach ought to differ as
little as possible from the investigative approach. This reduces the possibility that a common
element in the sample will have the same effect on both approaches. By using the statistical test,
we were able to determine whether the bias in the method being studied or the Undetermined
errors in the two approaches was to blame for the difference in the results.
Control determination: To reduce errors, a standard material is employed in experiments
under identical experimental conditions.
Dilution method: The dilution method is a vital technique for lowering interference errors. In
this procedure, dilution is carried out in a way that interferent species have little to no impact
below a specific concentration. This approach requires extreme caution because the sample is
diluted, and the dilution may change how the sample’s analyte is measured.
Standard addition: The known amount of standard solution of analyte is added to one portion
of the sample. The responses before and after the addition are measured and used to obtain the
analyte concentration. The standard addition method assumes the linear relationship between
response and analyte concentration.
Internal standard method: A known amount of reference species is added to all the samples,
standard, and blank. The response signal is obtained as the ratio of the analyte signal to the
reference species signal. It is utilized in chromatographic and spectroscopic analysis.
Parallel determination: To reduce the likelihood of unintentional errors, duplicate or triple
determination is performed instead of a single determination.
RANDOM DISTRIBUTION OF INDETERMINATE/RANDOM ERROR ACCURACY & PRECISION
Indeterminate errors (Random errors) :- Indeterminate errors represent random fluctuations in Accuracy and precision are the two important terminologies used in any measurement. These
measuring devices that are beyond the control of analyst. They are bidirectional. They are small two terms describe how closely a measurement resembles a standard or known value. Accuracy
in magnitude. But the accumulated effect can cause a considerable deviation to the mean. It is defines the degree to which a measurement is close to its true value. However, precision defines
not possible to determine the magnitude of individual random error since they are small in how consistently the same results are obtained from repeated measurements under the same
magnitude. circumstances. Measurements can be both precise and accurate, precise but not accurate,
Normal error curve and its properties: accurate but not precise, or neither. The in-depth discussion on accuracy and precision is
To understand the nature of random errors and to understand the development of normal error illustrated below:
curve, the following two points have to be considered. ACCURACY
1. In a particular measurement, assume that there are four small random errors (U1, U2, U3 and Accuracy indicates the closeness of the measurement to the true or accepted value. It is always
U4) combine to give an overall error. expressed by absolute and relative error. This measures the agreement between the result and
2. They have equal probability of occurring with a magnitude of deviation, ±U. The possible the accepted value. Accuracy is often more difficult to determine because the true value is really
combinations of the four random errors (U1, U2, U3 and U4) in a measurement are tabulated unknown. The accepted value must be used instead of the true value.
below. The comparison of two types of data is always made on the basis of an inverse measure of
accuracy which is an error. The smaller the error measured the greater the accuracy.
The accuracy has been classified into 3 types:
Point Accuracy: Point accuracy denotes that the instrument’s accuracy is only applicable to that
specific point on its scale. This kind of accuracy, however, says nothing about the instrument’s
overall accuracy. An excellent illustration of point accuracy is mass measurement.
Accuracy as Percentage of True Value: When the accuracy of the instrument is evaluated by
comparing the measured value to its true value, the result is expressed as a percentage of the
true value. Up to 0.5 percent of the true value is ignored due to instrument accuracy.
Accuracy as Percentage of Scale Range: The accuracy of a measurement is expressed as a
percentage of the scale range.
Expression of Accuracy
Absolute error: The absolute error E is the measurement of a quantity x which is given by
equation E = Xi – Xt Where, Xi = individual value, Xt = True or accepted value
Ex- Lets consider two numerical values 4.431 & 4.654, If 4.654 is regarded as true value then
Absolute error = Xi – Xt = 4.654 – 4.431 = 0.223
Relative error: it is more usefull quantity than the absolute error Er = (Xi – Xt) * 100% / Xi
= 0.223/4.654 * 100 = 4.791
Relative error is also expressed in part per thousand = 4.791 ppt
When we plot the relative frequency and the magnitude of error we get, Fig. A.
When the same procedure is applied for a measurement having ten uncertainties (U!, U2, U3, U4,
U5, U6, U7, U8, U9 and U10), then the shape of the curve will be similar (Fig. B).
PRECISION
Precision describes the reproducibility of measurements. In other words, precision can be
When the same procedure is applied for a very large number of individual random errors, a defined as the closeness of results that have been obtained in exactly the same way. The
smooth bell shaped curve is obtained (Fig. C). precision of a measurement can be easily measured by simply repeating the experiment to
obtain a replicated sample.
The precision of measurement is generally measured by the standard deviation, variance, and
coefficient of variation. These are functions of deviation from the mean. The precision can be
also expressed in terms of the range of data. It doesn’t tell anything about true value.
Precision includes two important terms:
When the same procedure is applied for a Repeatability: The fluctuation that results from taking multiple measurements under the same
very large number of individual random circumstances in a short period of time.
errors, a smooth bell shaped curve is Reproducibility: The variation appears when employing the same measurement technique with
obtained (Fig. C). When the deviation is various instruments, and operators, and over extended time periods.
replaced with the standard deviation (or The well-precise value may be inaccurate. The accurate value may not be precise sometimes.
true standard deviation) of the large The determinate type of error which lead to inaccuracy may or may not affect precision
number of individual measurements, then depending on how nearly it remains constant throughout that series of experiment. This
also the shape of the curve is the same (Fig. indicates accuracy and precision of data are not indicative of each other.
D) and such a plot is called normal error However, no matter how precise or accurate a measurement is, there is still some amount of error.
curve or Gaussian curve This is because the true value of a quantity cannot be measured with infinite precision as there are
always variations in measurements.
Error (E) is the difference between a measurement and the true value of the measurement (the
quantity being measured). Error is what causes values to differ when a measurement is repeated
and none of the results can be preferred over the others. Although it is not possible to
completely eliminate error in a measurement, it can be controlled and characterized. The total
error is usually a combination of systematic error and random error.
Random Errors These are fluctuations in the measured data due to the limitations of the
measurement device. Such errors results due to the experimenter's inability to take the
same measurement repeatedly in exactly the same way so as to get the exact value. Such errors
affect the precision of measurements and may be reduced by taking a number of reading and
using an average of the data set. The following variables are associated with random errors.
✓ Visual judgement with respect to reading and recording measurements on laboratory
equipment. Temperature fluctuations. This may affect the volume of the glassware, the viscosity
of a liquid to be measured or the performance of a balance.
✓ Wind that cause vibrations in the readings from a balance
Precision depends on the distribution of random errors since it is not based on the
true/accepted value.
Systematic Errors This is a consistent difference between a measurement and its true value
and persists throughout an entire experiment. Such errors affect all the data set in the same way
each time a measurement is made and cannot be reduced or eliminated by taking a number of
measurements and using an average of the data set. Systemic errors affect the accuracy of a
measurement and may arise due to:
✔ Instrument errors, these include:
Errors caused by an instrument being wrongly calibrated
■ Instruments being used under different conditions from which they were calibrated.
■ Instrument not reading zero when it should.
✓ Method errors, these include:
Incompleteness of reactions
■ The occurrence of side reactions cause by impurities
Properties of a normal error curve
1. Zero indeterminate error occurs with maximum frequency. ✓ Personal errors which result from personal judgement such as the end-point or titration
2. A symmetry about the maximum indicates that the negative and positive errors occur with reactions or estimating measurements between scale markings.
equal frequency. ACCURACY & PRECISION EXAMPLE
3. An exponential decrease in frequency occurs as the magnitude of the error increases. Imagine a football player aiming at the goal to get a concept of accuracy and precision. The
4. The distribution of the normal error curve (Fig. above), can be described mathematically in player is considered accurate if his shot finds the net. A football player who repeatedly strikes
terms of three parameters. the same goalpost is precise but not accurate. Therefore, if a football player strikes the ball all
over the place but still scores, he or she is accurate without being precise. Whether he/she
Y = [exp(-(x-μ)2/2σ2)]/σ(2π) ½ scores or not, a precise player will consistently strike the ball to the same spot. A precise and
x = values of individual measurements accurate football player will not only aim at a specific spot but also score a goal.
μ = Arithmetic mean for an infinite number of such measurements (true mean) Difference between Accuracy and Precision
σ = True standard deviation
Accuracy Precision
Accuracy defines the closeness of the measurement Precision defines the reproducibility of
to the true or accepted value measurements.
Measurement can be accurate but not necessarily Measurement can be precise but not
precise. necessarily accurate.
Indicates how closely the results match the Indicates how closely results agree with
reference value. one another.
Requires a number of measurements to
Can be calculated using just one measurement.
be calculated.
May be affected by the indeterminate
May be affected by the determinate error.
error.
RELIABILITY OF RESULT Significant Figures
In analytical chemistry Reliability is used to express the consistency, accuracy & When working with analytical data it is important to be certain that you are using and reporting
reproducibility of the measurement. The extent to which the result are consistent over time the correct number of significant figures. The number of significant figures is dependent upon the
and accurate representation of the total population under study is called as reliability. The uncertainty of the measurement or process of establishing a given reported value. In a given
reliability of measurement is expressed in statstistical term called as Q test. number, the figures reported, i.e. significant figures, are those digits that are certain and the first
The Q test is defined as uncertain digit. It is confusing to the reader to see data or values reported without the
Q = | Questionable value – nearest value | / | Largest value – Smallest value | uncertainty reported with that value.
When Q value is less than critical value then such results are accepted. Example: Thus in the quantities 1.2680 gm and 1.0062 gm the zero is significant, but in the
Q- Value < Critical value = Accepted. quantity 0.0025 kg the zeros are not significant figures; they serve only to locate the decimal
If Q value is greater than the critical value then such result results are not accepted. point and can be omitted by proper choice of units.
Q – Value > Critical Value = Result is not accepted Rules for deciding the number of significant figures in a measured quantity:
The value of Q-Test are compared with critical values given in the following table (1) All nonzero digits are significant:
1.234 g has 4 significant figures,
Where sample size = How many times we did our 1.2 g has 2 significant figures.
experiment / how many result we have taken in (2) Zeroes between nonzero digits are significant:
our experiment 1002 kg has 4 significant figures,
3.07 mL has 3 significant figures.
In a sample of the following values are obtained (3) Zeroes to the left of the first nonzero digits are not significant; such zeroes merely indicate
at 4.3, 4.1, 4.0 & 3.2 micro gram per milligram of the position of the decimal point
cadmium present, should last reading 3.2 is 0.001o C has only 1 significant figure,
rejected or accepted 0.012 g has 2 significant figures.
(4) Zeroes to the right of a decimal point in a number are significant:
0.023 mL has 2 significant figures,
Q = | Questionable value – nearest value | / | Largest value – Smallest value | 0.200 g has 3 significant figures.
Where Q.V. = 3.2 ( it is the value in which we have doubt to clear) (5) When a number ends in zeroes that are not to the right of a decimal point, the zeroes are not
Nearest Value = 4.0 ( Value nearest to Q.V. ) necessarily significant:
Largest Value = 4.3 190 miles may be 2 or 3 significant figures, 50,600 calories may be 3, 4, or 5 significant figures.
Smallest Value = 3.2 The potential ambiguity in the last rule can be avoided by the use of standard exponential, or
Putting all the values in above formula ”scientific,” notation. For example, depending on whether 3, 4, or 5 significant figures is correct,
Hence we get, we could write
Q = [ 3.2 – 4.0 / 4.3 – 3.2 ] = 0.8 / 1.1 = 0.727 50,6000 calories as:
Which Is the calculated critical value for sample size 4 = 0.727 5.06 × 104 calories (3 significant figures)
& table critical value for sample size 4 = 0.831 5.060 × 104 calories (4 significant figures), or
Calculated critical value < table critical value than the figure is retained not rejected . 5.0600 × 104 calories (5 significant figures).
0.727 > 0.831 Exact numbers
Result: 3.2 is not rejected. Some numbers are exact because they are known with complete certainty. Most exact numbers
are integers: exactly 12 inches are in a foot, there might be exactly 23 students in a class. Exact
numbers are often found as conversion factors or as counts of objects.
Exact numbers can be considered to have an infinite number of significant figures. Thus, number
of apparent significant figures in any exact number can be ignored as a limiting factor in
determining the number of significant figures in the result of a calculation.
Rules for rounding off numbers
(1) If the digit to be dropped is greater than 5, the last retained digit is increased by one. For
example, 12.6 is rounded to 13.
(2) If the digit to be dropped is less than 5, the last remaining digit is left as it is. For example,
12.4 is rounded to 12.
(3) If the digit to be dropped is 5, and if any digit following it is not zero, the last remaining digit
is increased by one. For example, 12.51 is rounded to 13.
(4) If the digit to be dropped is 5 and is followed only by zeroes, the last remaining digit is
increased by one if it is odd, but left as it is if even. For example, 11.5 is rounded to 12, 12.5 is
rounded to 12.
This rule means that if the digit to be dropped is 5 followed only by zeroes, the result is always
rounded to the even digit. The rationale is to avoid bias in rounding: half of the time we round up,
half the time we round down.
SIGNIFICANT FIGURES IN CALCULATION
When combining measurements with different degrees of accuracy and precision, the number of
significant digits in the final answer can be no greater than the number of significant digits in the
least precise measured value. There are two different rules, one for multiplication and division
and the other for addition and subtraction, as discussed below.
1. For multiplication and division: The result should have the same number of significant figures
as the quantity having the least significant figures entering into the calculation. For example, the
area of a circle can be calculated from its radius using A = πr2
. Let us see how many significant figures the area has if the radius has only two—say, r = 1.2 m.
Then,
A = π r2 = (3.1415927…) × (1.2 m)2 = 4.5238934 m2
is what you would get using a calculator that has an eight-digit output. But because the radius has
only two significant figures, it limits the calculated quantity to two significant figures or
A = 4.5 m2
even though πbis good to at least eight digits.
2. For addition and subtraction: The answer can contain no more decimal places than the least
precise measurement. Suppose that you buy 7.56 kg of potatoes in a grocery store as measured
with a scale with precision 0.01 kg. Then you drop off 6.052-kg of potatoes at your laboratory as
measured by a scale with precision 0.001 kg. Finally, you go home and add 13.7 kg of potatoes as
measured by a bathroom scale with precision 0.1 kg. How many kilograms of potatoes do you
now have, and how many significant figures are appropriate in the answer? The mass is found by
simple addition and subtraction:
7.56 kg − 6.052 kg + 13.7 kg = 15.208 kg = 15.2 kg
Next, we identify the least precise measurement: 13.7 kg. This measurement is expressed to the
0.1 decimal place, so our final answer must also be expressed to the 0.1 decimal place. Thus, the
answer is rounded to the tenths place, giving us 15.2 kg.