ANALYTICAL CHEMISTRY

BS CHEMISTRY
SEMESTER-IV

Compiled from Analytical Chemistry by GD Christian

 1. Introduction of Analytical Chemistry:
Analytical chemistry is concerned with the chemical characterization of matter and
the answer to two important questions: what is it (qualitative analysis) and how
much is it (quantitative analysis).

2. Application of Analytical Chemistry

Analytical chemistry plays an important role in nearly all aspects of chemistry, for
example, agricultural, clinical, environmental, forensic, manufacturing,
metallurgical, and pharmaceutical chemistry

 The nitrogen content of a fertilizer determines its value & nitrogen content
can be determined by analytical chemistry. So Analytical Chemistry has its
application in Agriculture Sciences.

 Foods must be analyzed for contaminants (e.g., pesticide residues) and for
essential nutrients (e.g., vitamin content). For this purpose, Analytical
Chemistry is used.

 The air we breathe must be analyzed for toxic gases (e.g., carbon monoxide).
So, Analytical chemistry will be used in environmental sciences for the
determination of toxic gases.

 Blood glucose must be monitored in diabetics (and, in fact, most diseases are
diagnosed by chemical analysis). In this way, Analytical Chemistry has its
applications in clinical labs.

Analytical chemists determine what and how much. Analytical chemists serve the
needs of many fields.

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  In medicine, analytical chemistry is the basis for clinical laboratory tests
which help physicians diagnose disease and chart progress in recovery.
 In industry, analytical chemistry provides the means of testing raw
materials and for assuring the quality of finished products whose chemical
composition is critical. Many household products, fuels, paints,
pharmaceuticals, etc. are analyzed by the procedures developed by analytical
chemists before being sold to the consumer.
 Environmental quality is often evaluated by testing for suspected
contaminants using the techniques of analytical chemistry.
 The nutritional value of food is determined by chemical analysis for major
components such as protein and carbohydrates and trace components such as
vitamins and minerals. Indeed, even the calories in food are often calculated
from its chemical analysis.
 Analytical chemists also make important contributions to fields as diverse as
forensics, archaeology, and space science.

3. QUALITATIVE & QUANTITATIVE ANALYSIS:

The discipline of analytical chemistry consists of qualitative analysis and
quantitative.

 Qualitative Analysis:

Qualitative analysis tells us what chemicals are present. It is the Identification of

elements, ions, or compounds present in a sample (we may be interested in whether
only a given substance is present).

 Quantitative Analysis:

Quantitative analysis tells us how much. Determination of how much of one or

more constituents is present is called quantitative analysis.
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Example: The sample may be solid, liquid, gas, or a mixture. The presence of
gunpowder residue on a hand generally requires only qualitative knowledge, not of
how much is there, but the price of coal will be determined by the percent of
undesired sulfur impurity present.

Qualitative tests may be performed by selective chemical reactions or with the use
of instrumentation. The formation of a white precipitate when adding a solution of
silver nitrate in dilute nitric acid to a dissolved sample indicates the presence of a
halide. (Precipitation Test).

Certain chemical reactions will produce colors to indicate the presence of classes of
organic compounds, for example, ketones. (Solubility Test).

A clear distinction should be made between the terms selective and specific

 A selective reaction or test is one that can occur with other substances but
exhibits a degree of preference for the substance of interest.
 A specific reaction or test is one that occurs only with the substance of
interest.

Unfortunately, very few reactions are truly specific but many exhibit selectivity.

Selectivity may be also achieved by a number of strategies. Some examples are:

 Sample preparation (e.g., extractions, precipitation)

 Instrumentation (selective detectors)
 Target analyte derivatization (e.g., derivatize specific functional groups)
 Chromatography, which separates the sample constituents.

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 4. STEPS OF CHEMICAL ANALYSIS

Step # 1 Define a Problem

Before the analyst can design an analysis procedure, he or she must know what
information is needed, by whom, for what purpose, and what type of sample is to
be analyzed. So, first step in the process of chemical analysis is to define the
problem. Analyst should have good communication with client.

First of all, analyst will find out that either client want quantitative analysis or
qualitative analysis of his sample. Then after communication, he will define the
problem by asking to client that how much accurate, sensitive & precise results he
wants & what will be the budget of analysis of sample.

The analyst (the problem solver) should consult with the client to plan a useful
and efficient analysis, including how to obtain a useful sample.

Once the problem is defined this will dictate how the sample is to be obtained, how
much is needed, how sensitive the method must be, how accurate and precise it
must be, and what separations may be required to eliminate interferences. So,
Analyst will move to next step in the process of chemical analysis.

Step # 2 Select a Method

Once the required measurement is known, the analytical method to be used will
depend on a number of factors, including the analyst’s skills and training in

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different techniques and instruments; the facilities, equipment, and instrumentation
available; the sensitivity and precision required; the cost and the budget available;
and the time for analysis and how soon results are needed.

Step # 3 Obtaining a Representative Sample

The material to be sampled may be solid, liquid, or gas. It may be homogeneous or

heterogeneous in composition.

 For Homogeneous Sample: If sample will be homogeneous than sample

can be obtain by grab sampling. In this process, a simple “grab sample”
taken at random will suffice for the analysis.

 For Heterogeneous Sample: There may be variation in composition

throughout the sample, in this case several individual samples will be
required. This process of obtaining several individual samples is called gross
sampling.

If the gross composition is needed, then special sampling techniques will be

required to obtain a representative sample.

For example, in analyzing for the average protein content of a shipment of grain, a
small sample may be taken from each bag, or tenth bag for a large shipment, and
combined to obtain a gross sample.

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Sampling is best done when the material is being moved, if it is large, in order to
gain access.

Laboratory Sample & Analysis Sample

The larger the particle size, the larger should be the gross sample. The gross
sample must be reduced in size to obtain a laboratory sample of several grams,
from which a few grams to milligrams will be taken to be analyzed (analysis
sample).

The size reduction may require taking portions (e.g., two quarters) and mixing, in
several steps, as well as crushing and sieving to obtain a uniform powder for
analysis.

In the case of biological fluids, the conditions under which the sample is collected
can be important, for example, whether a patient has just eaten. The composition of
blood varies considerably before and after meals, and for many analyses a sample
is collected after the patient has fasted for a number of hours. Persons who have
their blood checked for cholesterol levels are asked to fast for up to twelve hours
prior to sampling.

Preservatives such as sodium fluoride for glucose preservation and anticoagulants

for blood samples may be added when samples are collected; these may affect a
particular analysis.

Certain precautions should be taken in handling and storing samples to prevent or

minimize contamination, loss, decomposition, or matrix change. In general, one
must prevent contamination or alteration of the sample by the;

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 1. Container: Trace constituents may be lost during storage by adsorption onto
the container walls.
2. Atmosphere: The sample may have to be protected from the atmosphere or
from light. It may be an alkaline substance, for example, which will react
with carbon dioxide in the air. Blood samples to be analyzed for CO2 must
be protected from the atmosphere.
3. Heat/Temperature: Sample if protein or enzyme in nature than it may
degrade with the increase of temperature. So optimal temperature is required
for sample preservation.
4. Light.

Step # 4 Prepare the Sample for Analysis

• Measure the size of sample:

The first step in analyzing a sample is to measure the amount being analyzed (e.g.,
volume or weight of sample). This will be needed to calculate the percent
composition from the amount of analyte found. The analytical sample size must be
measured to the degree of precision and accuracy required for the analysis.

• Drying of solid sample

Solid samples are often analyzed on a dry basis and must be dried in an oven at
110 to 120.C for 1 to 2 h and cooled in a dessicator before weighing, if the sample
is stable at the drying temperatures.

Some samples may require higher temperatures and longer heating time (e.g.,
overnight) because of their great affinity for moisture.

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The amount of sample taken will depend on the concentration of the analyte and
how much is needed for isolation and measurement. Determination of a major
constituent may require only 100 mg of sample, while a trace constituent may
require several grams.

• Dissolution of solid sample in solution form

More often, the sample must be in solution form for measurement, and solids must
be dissolved. Inorganic materials may be dissolved in various acids, redox, or
complexing media. Acid-resistant material may require fusion with an acidic or
basic flux in the molten state to render it soluble in dilute acid or water.

• Dry Ashing:

Ashing is the burning of organic matter. Organic materials that are to be analyzed
for inorganic constituents, for example, trace metals, may be destroyed by dry
ashing. The sample is slowly combusted in a furnace at 400 to 700.C, leaving
behind an inorganic residue that is soluble in dilute acid.

• Wet Digestion

Digestion is the wet oxidation of organic matter. The organic matter may be
destroyed by wet digestion by heating with oxidizing acids. A mixture of nitric and
sulfuric acids is common. Perchloric acid digestion is used for complete oxidative
digestion.

Biological fluids may sometimes be analyzed directly. Often, however, proteins

interfere and must be removed. Dry ashing and wet digestion accomplish such
removal.

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For Organic Analyte:

If the analyte is organic in nature, these oxidizing methods cannot be used. Rather,
the analyte may be extracted away from the sample or dialyzed, or the sample
dissolved in an appropriate solvent. It may be possible to measure the analyte
nondestructively. An example is the direct determination of protein in feeds by
near-infrared spectrometry.

Once a sample is in solution, the solution conditions must be adjusted for the
next stage of the analysis (separation or measurement step).

For example, the pH may have to be adjusted, or a reagent added to react with and
“mask” interference from other constituents. The analyte may have to be reacted
with a reagent to convert it to a form suitable for measurement or separation.

Blank Solvent (Blank Determination)

The solvents and reagents used for dissolution and preparation of the solution
should be of high purity (reagent grade). Even so, they may contain trace
impurities of the analyte. Hence, it is important to prepare and analyze replicate
blanks, particularly for trace analyses.

A blank theoretically consists of all chemicals in the unknown and used in an

analysis in the same amounts (including water), run through the entire analytical
procedure. The blank result is subtracted from the analytical sample result to arrive
at a net analyte concentration in the sample solution.

Step # 4 Perform Necessary Chemical Separation

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Analyst must perform more than one separation steps in order to:

 Eliminate interferences.
 To provide suitable selectivity in the measurement,
 To pre concentrate the analyte for more sensitive or accurate measurement.
 Seaparation of analyte from matrix is necessary to avoid error in result.

Separation steps may include:

1) Precipitation, 2) Extraction into an immiscible solvent,

3) Chromatography, 4) Dialysis, and Distillation.

Step # 5 Performing the Measurements

The Method employed for the actual quantitative measurement of the analyte will
depend on a number of factors which are following:

a) Selectivity, b) Sensitivity, c) Accuracy, d) Precision,

e) Cost, and f) Rapidity.(Speed)

Analytical chemistry research often deals with the optimization of one or more of
these parameters, as they relate to a particular analysis or analysis technique.

1. Classical Methods for Measurements:

a. Gravimetric Analysis

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Gravimetric analysis usually involves the selective separation of the analyte by
precipitation, followed by the very nonselective measurement of mass (of the
precipitate)

b. Volumetric or titrimetric analysis (Titration)

Volumetric, or titrimetric, analysis, the analyte reacts with a measured volume of

reagent of known concentration, in a process called titration.

• Gravimetric and volumetric analyses can provide results accurate and

precise to a few parts per thousand (tenth of 1 percent) or better.

• However, they require relatively large (millimole or milligram) quantities of

analyte and are only suited for the measurement of major constituents,

• Volumetric analysis is more rapid than gravimetric analysis and is therefore

preferred when applicable.

Modern Methods for Measurements:

Instrumental Techniques:

• Instrumental techniques are used for many analyses (instrumental analysis).

• Based on the measurement of a physical property of the sample,

For example, an electrical property or the absorption of electromagnetic radiation.

Examples are spectrophotometry (ultraviolet, visible, or infrared), atomic
spectroscopy (absorption, emission), mass spectrometry, nuclear magnetic
resonance spectrometry (NMR), X-ray spectroscopy (absorption, fluorescence),

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electroanalytical chemistry (potentiometric, voltammetric, electrolytic),
chromatography (gas, liquid).
Advantages:

• Instrumental techniques are generally more sensitive and selective than the
classical techniques but are less precise, on the order of 1 to 5% or so.

• These techniques are usually much more expensive, especially in terms of

initial capital investment. But depending on the numbers of analyses, they
may be less expensive when one factors in personnel costs.

• They are usually more rapid, may be automated, and may be capable of
measuring more than one analyte at a time.

Chromatography techniques are particularly powerful for analyzing complex

mixtures.

• Constituents are separated as they are pushed through (eluted from) a

column of appropriate material that interacts with the analytes to varying
degrees, and these are sensed with an appropriate detector as they emerge
from the column, to give a transient peak signal, proportional to the amount
of each.

Some errors may occur during measurement of results. These errors may be due to
some fault in instrument or inexperience of analyst. Errors in measurements of
results may occur if method of analysis is not proper. So, it is necessary to
minimize these errors. In this way, our result of chemical analysis will be reliable
& exact.

Minimize Errors in Measurements/ Result:

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First major cause of error in analytical techniques is due to faulty instruments,
glassware or apparatus. So there are some methods to identify these errors &
correct these errors.

 Calibration:

If any determinate error is present in our measurements due to faulty instruments

than it can be identified & removed by Calibration.

So, 1st method to minimize error is calibration of instruments or Apparatus.

Periodic Calibration of instruments will help us to minimize the error.

Calibration is accomplished by preparing a series of standard solutions of the

analyte at known concentrations and measuring the instrument response to each of
these (usually after treating them in the same manner as the samples) to prepare an
analytical calibration curve of response versus concentration.

Running Blank Determination

In this method, we will run blank determination & it also helps to minimize error
of instrument.

In blank determination we donot take analyte, we will only take reagent & solvent.
So, we can determine impurities present in the reagent or solvent.

In this way, value of those impurities can be substracted from the result & in this
way error can be reduced.

Example:

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Normal Determination- Analyte + Solvent + Reagent

Blank Determination- Solvent + Reagent (No Analyte)

So, If any error is found in result due to reagent or solvent than it can be identified
by blank determination. Reading of blank determination is taken into consideration
for minimizing the error in normal determination.

Controlled Determination:

Same reagent & solvent is used that we are using in normal determination but in
this determination, we take a standard substance.

Standard substance ( Known concentration/ amount) of analyte is taken &

analyzed, compared with normal determination.

Normal Determination- Analyte + Solvent + Reagent

Controlled Determination- Standard + Solvent+ Reagent

Standard ADDITION METHOD:

Standard substance of known amount of analyte is added.

Sample + Reagent 10 µg (After Analysis)

Standard (5µg) + Reagent + Sample (10µg) 15 µg (After Analysis)

The sample matrix may affect the instrument response to the analyte. In such cases,
used to overcome sample matrix calibration may be accomplished by the method
of standard additions. The sample is spiked with a known amount of standard, and

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the increase in signal is due to the standard. In this manner, the standard is
subjected to the same environment as the analyte.

Internal Standard Addition method:

An instrumental response is often subject to variations from one measurement to

the next due to changing instrument conditions, resulting in imprecision

In standard Addition we are taking same standard as present in analyte but in

Internal standard addition method, we are using different internal standard which
will not be similar to analyte.

Suitable internal standard is selected & analyzed in same experimental conditions.

Step # 6 CALCULATING THE RESULTS AND REPORTING THE DATA

Once the concentration of analyte in the prepared sample solution has been
determined, the results are used to calculate the amount of analyte in the original
sample.

 Result should express accuracy.

 Replicate analyses can be performed (three or more), and a precision of the
analysis may be reported,

 The analyst should critically evaluate whether the results are reasonable and
relate to the analytical problem as originally stated

Remember that the customer often does not have a scientific background so will
take a number as gospel. Only you, as analyst, can put that number in perspective,
and it is important that you have good communication and interaction with the
“customer” about what the analysis represents.

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 Electro-analytical Methods:

 Volatametry (Measure of voltage by voltmeter),

 Potentiometry (Measurement of potential difference)
 Ammetry (Measure current.

Spectroscopic Techniques: UV Visible Spectroscopy, Atomic Absorption & atomic

emission spectroscopy,NMR, IR spectroscopy.

Separation Methods:

Solvent Extraction, Chromatography, distillation

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 CLASSIFICATION OF ANALYTICAL TECHNIQUES

Instrumental Methods
CLASSICAL METHODS

Volumertric/Titrimetric
Gravimetric/ Electro-analytical
Spectroscopic
Method Techniques
Techniques
Gravimertry

Separation Techniques

Solvent Extraction,
Chromatography,
distillation.

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ERRORS
Analytical chemistry is based on reliability, reproducibility and
accuracy. However, every measurement has some degree of uncertainty
which is called as error in analytical chemistry.

“The difference between the experimental mean value and a true

value is called ERROR”

Error is the difference appear between measured value and expected

value or can be calculated by subtracting the expected value from
measured value or vice versa to know how much error is present.

Classification:

Errors are classified into two types – determinate and indeterminate

errors.

1. Determinate/Systemic error is the error which can be determined at

any stage and can be rectified at once.

Sources/Classification of Determinate ERROR:

 Instrumental Error

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Errors occur due to faulty instrument or reagent containing impurities
e.g. un-calibrated weights, un-calibrated burette, pipette and measuring
flasks.

 Error of Method

When errors occur due to method, it is difficult to correct. In

gravimetric analysis, error occurs due to Insolubility of precipitates,
co-precipitates, post-precipitates, decomposition, and volatilization.

 Operational/Personal Errors:

Errors for which the individual analyst is responsible and are not
connected with the method or procedure is called as personal errors e.g.
unable to judge color change

 Additive Error

Additive error does not depend on constituent present in the

determination e.g. loss in weight of a crucible in which a precipitate is
ignited.

 Proportional Error

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Proportional error depends on the amount of the constituent e.g.
impurities in standard compound.

Determinate errors can be avoided or corrected.

Systemic/ Determinate errors are caused by experimenter carelessness or

equipment failure.

Errors cannot be neglected but minimized by

 Calibrating instruments,
 Running blank determination and control determination,
 Standard addition,
 Internal standard

2. Indeterminate error/Random Error: is the error which is difficult

to determine or indefinite.

Random (or indeterminate) errors are caused by uncontrollable

fluctuations in variables that affect experimental results. Indeterminate
errors are random errors over which analyst has no control.

Determinate error can appear again and again but can be corrected
whereas indeterminate error appears rarely and not get eliminated.

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