INTRODUCTION TO ANALYTICAL CHEMISTRY

LESSON ONE

Analytical Chemistry

1.0 Introduction

In this lesson, we shall begin by making known to you what analytical chemistry is. The question
of what, where, who and why of analytical chemistry will also be answered in this lesson. It will
be interesting to learn that analytical chemistry is all around us and everything we do has an
aspect of analytical chemistry.

1.1 Objectives

 By the end of this lesson the learner should be able to:

(i) State the importance and application of analytical chemistry

(ii) Classify the methods of analysis
(iii) Differentiate between classical and instrumental methods of analysis
(iv) State the factors to be considered in selecting analytical methods
(v) State some problems associated with trace analysis.

1.2 Nature, Scope and Importance of Analytical Chemistry

The word analysis originates from two Greek words: ana meaning up and lyein meaning loosen.
Analysis means to loosen up or separate into parts. Therefore analysis can be defined as the
identification and determination of the relative amounts of one or more components in a mixture.
Identification deals with the qualitative measurement while determination deals with the
quantitative measurement of one or more components in a mixture. Analytical chemistry tries to
answer the questions what is in a sample, that is the sample components and how much of the
component is in the sample. It also tries to develop new methods or techniques for qualitative
and quantitative measurement. However the primary objective of analytical chemistry is to gain
qualitative and quantitative information about the composition of an analytical sample. An
analytical sample may be biological such as blood and urine or environmental such as water and
soil samples.

 Name any other five examples of samples

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NOTE:

 There are two major areas of analytical chemistry are:-

 Qualitative analysis: where the constituents of an analytical sample are determined. The
constituents in a sample are usually unknown or suspected so that analysis is done to reveal
them.

 Quantitative analysis: where the amount of the constituent in an analytical sample is

determined. The constituents are known or suspected to be present so that the exact amount is
determined.

 It is therefore important to understand the analytical procedures that are the basis of the
quantitative and qualitative determination.

? Question: - What are the concerns of analytical chemistry?

1.3 Position of analytical chemistry and its application

Traditionally chemistry could be divided into five areas; physical, organic, inorganic
biochemical and analytical. Biochemistry has however become a branch of its own in many
institutions, and another area of chemistry, theoretical, has come up. Here analytical chemistry
could be thought of as a separate area or branch of chemistry. Nowadays these divisions in
chemistry are hard to make because of the crossovers from one area to another. For example
analytical chemistry is needed in all the areas of chemistry. Further analytical chemistry is
needed in other scientific and technical disciplines. This has enabled the advancement of the
analytical chemistry, which in turn has resulted into the development of new areas of science
such as chemistry physics, biophysics and molecular biology.

Analytical chemistry has therefore a multidisciplinary application. It is applied in many fields such as in
medicine (clinical chemistry, in agriculture (soil, plants analysis), in the environment (pollution of air,
water and soil). This makes the areas of interest of analytical chemistry many and diverse. Some more
areas where analytical chemists are involved are as shown in Figure 1 .

Figure 1: The various areas where an analytical chemistry serves

1.4 Role of an analytical chemist

The sample gives a guide on the type on the type of analysis to be done to extract the required
information. Therefore the roles of an analytical chemist are vast. Among them is one or more of the
following:-

(i) Instrumental design

(ii) Sampling techniques
(iii) Development and evaluation of new procedures
(iv) Laboratory management
(v) Classical analysis
(vi) Statistical analysis
(vii) Optical systems development
(viii) Computer programming and electronics.

 So you as an analytical chemist have a wide opportunity to work! See the following note.

NOTE:

 Specific examples of areas in which analytical chemistry is involved include:-

 Economic value:- determining the amount of silver or gold in a coin or amount of

protein in animal feed, the amount of vitamin or aspirin in a tablet.
 Health hazard: - determining the amount of CO,SO 2 in emission from charcoal burning,
amount of lead from a car exhaust.
 Disease diagnosis: - determining glucose in urine for diabetics.
 Quality control: - drugs and food manufacturers determine quality of starting material
and finished products to make sure stated standards are maintained.
 Relating properties to composition and structure: - both physical and chemical
properties of alloys adhesives, lubricants, plastics depend on their chemical composition.
Also activity of pesticides, herbicides, drugs depend on their structure.
 Research activities: - determining accumulation and fate of pesticide in the food chain
or metabolic pathways of drugs in the human body, looking for way to separate, identity
and determine new products, enzymes, and proteins etc. developing new and better
instruments for carrying out qualitative measurements.

 Think of yourself as an analytical chemist and write a sentence about where and what
you would like to be and do respectively.
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1.5 Methods of Analysis

As mentioned earlier there are two basic aspects of analysis that is, the identification of
constituents in a sample and estimation of the amount of each individual constituent. The
identification aspect involves sampling, separation of interfering substances and identification or
analysis, while the estimation aspect goes further to measure the desired constituent and the
calculation and interpretation of data. Qualitative analysis methods can deal with partial analysis
where only some components are determined or complete analysis where all substances in the
sample are analyzed.

The methods used in estimation or determinations are classified depending upon:-

i) The instrument/equipment used. Here the methods are further classified as either
classical (chemical or wet) or as modern (instrumental) methods.

ii) The sample size. Here the methods are either macro or micro analysis.

Types of analysis

1.5.1 Classical (chemical or wet) methods

These are the methods in which basic equipments are used based on simple chemical analysis.
Methods under this category include volumetric and gravimetric analysis. The methods usually
require samples with high concentrations. In this module these methods will be learnt.

1.5.2 Modern (instrumental) methods

These are methods which are based on measurement of physical or chemical property of a
substance and use of electronic, thermal or optical principles for determination. These methods
can be used to analyze samples with low concentrations. Many advances in analytical
measurements have occurred through modern methods; although a combination of the methods is
normally applied. These methods will be handled in higher level modules.

A summary of the analytical techniques in this classification is shown in the table 1.

Table 1: A summary of classical and instrumental techniques

Basis of Method Classification Example

Titrimetric classical Acid/base, redox, precipitation titrations

Gravimetric classical Analysis of inorganic ions (constant mass)

Separation classical TLC, PC,CC, IEC

 instrumental GLC, HPLC

Electrochemical classical Potentiometric, conductimetric titrations

instrumental Polarography, ion selective electrodes

Spectroscopy – absorption instrumental UV, IR, NMR, ESR, AAS

Spectroscopy-emission Flame photometry, AES, MS

Nuclear/X-ray instrumental MS (isotopes), X-ray fluorescence, NA

Thermal instrumental DTA, TGA, DSC

Combination Techniques instrumental GS/MS, GC/IR, HPLC/MS

Key: TLC – Thin Layer Chromatography, PC – Paper Chromatography, CC – Column

Chromatography, IEC – Ion Exchange Chromatography, GLC – Gas Liquid Chromatography,
HPLC – High Pressure (performance) Liquid Chromatography, UV – Ultraviolet, IR – infrared,
NMR – Nuclear Magnetic Resonance, ESR – Electron Spin Resonance, AAS – Atomic
Absorption Spectroscopy, AES – Atomic Emission Spectroscopy, MS – Mass Spectroscopy, NA
– Neutron Activation, DTA – Differential Thermal Analysis, DSC – differential scanning
colorimetry, TGA – Thermogravimetric Analysis

 Choose either classical or an instrumental technique you may be familiar with and
briefly describe it.
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1.5.3 Comparison of classical and instrumental methods

A method can be classified as either classical or instrumental depending on the procedure used.
For example in determination of the concentration of hydrochloric acid using sodium hydroxide,
where sodium hydroxide is added until the reaction is complete, thus the acid has been
neutralized. A visual aid such as formation of color is used to determine the end point then, the
method is termed as classical. However, if a pH meter is used to determine the end point then the
procedure would be termed instrumental. In classical methods, no mechanical or electrical
instruments are used except simple apparatus. Chemical reactions are used to quantify the
analyte of interest. Instrumental methods of analysis depend on instruments to carry
measurements or to perform the entire analysis. A brief comparison of the classical method (wet
chemistry) of analysis versus the instrumental methods is shown in table 2.

Table 2: Comparative analysis of classical and instrumental methods

Classical Instrumental
Accounts for about 10% of all the current Accounts for 90% of the current analytical
analytical work WHY? work WHY?
 Less sensitive (upto microgram units)  Very sensitive
 Determines one analyte at a time  May determine several samples
 Small concentration range of 2-3 compounds (multi-analyzer, even>10)
orders of magnitude i.e. powers of 10  More than 6 orders of magnitude i.e.
 Labor intensive power of 10
 Less efficient in data handling for  Quicker and cheaper for large samples
example in recording hence, less reliable  More efficient thus modern equipment
data. have automatic data handling hence more
reliable

Advantages Advantages
Simple procedures Automated equipment makes data/statistical
Procedures are often accurate handling easy
Method based on absolute measurements Require small volume of solution
Specialized training not required
Cheaper equipment

Disadvantages Disadvantages
Storage of large volume of solution Expensive equipment
Instability of some reagents Complex procedures
Need clean glassware Require skillful operator

 What therefore are any two advantages of titrimetric and gravimetric analysis over GLC?
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1.5.4 Analytical methods based on sample size

This classification depends on the minimum size of the analyte in the sample: here the methods
can be classified as macro or micro methods depending on the sensitivity of instrument. Table 3
shows some of the levels that can be analyzed depending on the sensitivity of the instrument.
Table 3: Classification of analytical methods by sample size

Method Minimum size Gram

Macro 100mg 10-1
Semi macro 10mg 10-2
Micro 1mg 10-3 miligram
Ultra micro 1ug 10-6 microgram
Sub microgram 10-2ug 10-8
10-9 nanogram
10-12 picogram
10-15 femtogram
10-18 attogram

1.5.3.1 Problems of trace analysis

Analysis at microgram level and below is normally called trace analysis. Such analysis faces
many problems some of these being:

(i) The interferences. There are other compounds present in the sample and can be detected
by the method used giving erroneous results. These should be removed or eliminated
before determination. For example in determination of lead using dithizone, presence of
other elements such as zinc, cadmium would also react with dithizone to obtain
inaccurate results. These elements therefore need to be removed.
(ii) Contamination is the presence of a minor and unwanted constituent in a material, physical bodyn
or natural environment.
a) Use of specific containers for specific compounds such as polythene bottles for mineral ions
or glass bottles for organic compounds are used.
b) Carrying out blank analysis helps to minimize contaminations from reagents and working
apparatus. A blank is a solution or extract that contains all other reagents and use similar
apparatus but does not contain the analyte (the substance being analysed). By comparing
the analysis of the sample and the blank one is able to obtain the correct concentration in
the sample. For example, in determining copper in a sample, the exact concentration { }
is obtained by {Cu} ={sample} – {black}.
c) Reagents are supplied with specific standards that are meant to reduce or estimate
contamination. International bodies such as environmental protection agency (EAP) of USA and
national physical laboratory (NPL) of UK provide standards for reagents.
(iii) Sample loses. This is felt more especially when dealing with low concentration samples.
Some amounts are usually lost through sticking on the wall of containers, especially
organic materials where upto 70% can be lost on the surface of the container.

1.5.5 Criterion for selection of methods of Analysis

The selection of the method to be used in a particular analysis depend on a number of factors.
Some of these include speed (the time required to carry out the analysis), for example if one has
to analyze many samples for a short time, one would choose a method that is fast, availability of
equipment in the field or laboratory, sample size available, concentration of material being
analyzed, nature of sample (its solubility, toxicity, radioacitivity, value), accuracy and precision
of data required, cost of analysis and identity of sample (whether to use destructive or non-
destructive method).

 What would you consider if you were to analyse for pollution of the main source of water
in your village? Explain your answer
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Summary

 In this lesson, we have defined analytical chemistry and explored its importance in
science and technology. A survey of the methods of analysis is indicated together with their
classification. Lastly some of the factors considered when selecting an analytical method have
been mentioned.

COLLECTION AND TREATMENT OF ANALYTICAL DATA

LESSON TWO

Steps of Analysis

2.0 Introduction
Any analysis requires five major steps; sampling, sample pre-treatment, conversion to desired
constituents to measurable form, measurement of desired constituent, and calculation and
interpretation of data. In this lesson we shall discuss these steps.

2.1 Objectives

 By the end of this lesson the learner should be able to:

(i) Outline the steps involved in obtaining analytical data

(ii) Describe the steps involved in obtaining analytical data

2.2 Sampling

A sample is a subset of a population that is used to represent the entire group as a whole. This implies
then that sampling is a statistical method of obtaining representative data for observations from a group
or population.

Sampling is an essential step in analysis and its main objective is to obtain a sample (a portion of
the substance analyzed) whose composition both qualitatively and quantitatively is representative
of the substance being sampled. However, before starting the sampling procedure it is important
to get information on the sample and the method to be used in analysis.

2.2.1 Factors to consider in sampling

This involves literature search on the sample and methods of analysis. The selection of the
method of analysis depends on the factors discussed in lesson one. The possibility of whether
impurities need to be separated before quantification is also explored. The frequency and
duration of sampling, sampling devices, sample size and the storage containers are also
considered in obtaining a representative and reproducible sample. Some samples give
measurements for a particular period.

For example in an air lead pollution analysis, it is important to collect sample during the rush
hour and between the rush hours in order to get a realistic indicator of the overall lead exposure.
The sampling equipment varies a lot for gas, liquid or solids samples, bottles for water sample,
corer for sediment samples, collector sampling device for air samples. The storage containers
should be such that the analytical sample does not react with it or with other constituents.

 What time will consider going to sample the source of water we addressed in lesson one?

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2.2.2 Methods of sampling

An analytical sample is a representative of the entire substance being analyzed. In a homogenous

substance, any portion of it forms a sample. Hence obtaining a representative sample is easy, by
taking a portion from it. However, actual biological or environmental samples are heterogeneous
and thus sampling is done carefully. Though different techniques are employed for sampling
gases, solids and liquids, there are two basic methods of obtaining representative samples:
random and statistical sampling. The method of sampling depends on the mass of sample
required, nature and size of batch sampled.

2.2.2.1 Random sampling

In random sampling a sample is taken in such a way that all portions of the substance have an
equal chance of being included. This can be done either by selecting population members at
regular intervals or by use of random number table where each member of the substance is
allocated a number and the numbers are picked randomly. This is done to avoid human bias.
Random sampling is used for materials that are discrete, such as tablets in pharmaceutical
industry.

Activity

 Explain how you would obtain paracetamol tablets from the industry using random
sampling.

 Name two other samples that can be sampled by random sampling

2.2.2.2 Statistical sampling

Statistical sampling follows a statistically based plan that gives every portion of the substance an
equal chance of appearing in the sample. This sampling is done on materials that are not discrete
but that are bulk. Basically two steps are involved in this kind of sampling.

a) Collection of a gross sample, with its size varying from a few grams or less to several
kilograms. The size of the gross sample is determined by the uncertainty that can be
tolerated between composition of the sample and the whole, the degree of heterogeneity
of the material being sampled, and the level of particle size at which heterogeneity
begins.

b) Reduction of the gross sample to a size convenient for laboratory work through blending,
grinding and many other techniques.

Activity

 Explain how you would obtain a sample of maize from the maize store harvested by a
large scale farmer using statistical sampling.

 Name any two samples that can be sampled by statistical sampling

2.3 Sample pre-treatment

Analytical samples are carefully handled to minimize contamination (or alteration), loss and
decomposition or matrix change from atmosphere exposure during storage. For example,
alkaline samples react with CO 2 in the air when not well protected; for metal analysis in water
samples these metal may be absorbed into the walls of the container if not acidified immediately.
Stability of the sample therefore should be considered before sampling. The samples may also be
stored under refrigeration as a mode of handling to prevent further action like bacterial activity.

A given quantity of the sample is then treated in different ways depending on its nature. For
example, solid samples may be dried in an oven and ground before putting into solution form
and perhaps removing interferences. Samples may undergo various treatment techniques such as
acid digestion to destroy other interferences or sometimes analyzed directly. These treatments
are meant to get the sample in a form suitable for measurement and separate the analyte from
interfering constituents. Some of the separation mechanisms include precipitation, extraction,
solubility, volatility, freezing and concentration. In this module some separation techniques will
be learnt.

 Which is the stable food in your locality? How do you preserve it? Would you describe
this method of preservation as a pretreatment?
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 You probably are sensing that analytical chemistry is in our daily lifes?

2.4 Methods of converting analytes to measurable form (Sample Preparation)

Generally, there are two methods of sample preparation:-

(iv) Totally destroying the sample matrix

This method is for inorganic analyte or those which can be converted to an inorganic derivative
for measurement. For example, in acid treatment, hydrochloric acid is a good general solvent for
dissolving metals that are above hydrogen in the electromotive series; nitric acid is a strong
oxidizing acid and will dissolve most of the common metals, nonferrous alloys and the ‘acid-
insoluble’ sulfides; perchloric acid dissolves most of the common metals and destroys traces of
organic matter.

Animal and plant tissue, biological fluids and organic compounds are usually decomposed by
wet digestion using a mixture of acids or by dry ashing at a high temperature 400 0C to 700 0C.
Here the mixture of acids oxidize organic matter to carbon dioxide, water and other volatile
products, which are driven off, leaving behind salts or acids of the inorganic constituents. In dry
ashing atmosphere oxygen serves as the oxidant where the organic matter is burned off, leaving
an inorganic residue.

 List three inorganic analytes.

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Further reading

 Acid digestion and dry ashing

(v) Non-destructive or only partially destructive

This is a common method for organic analytes. An appropriate reagent is added to the sample
and then with through grinding, mixing and refluxing desired constituents of solid materials
are extracted. This is done by using solvents that extract only certain groups of organic
compound in the sample.

 List three organic analytes.

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2.5 Measuring the Analyte

Measuring of the analyte can be done using wet/chemical or instrumental methods as was
mentioned in the first lesson. There are a number of instrumental methods of analysis available,
some of which have been outlined in table 1. The choice of the methods is determined by the
factors discussed earlier. The methods of choice are those that are able to indicate the presence of
the analyte (identify) as well as the amounts present (quantity). Before measurement, any
substance that may be present, which can interfere with the analysis may be removed or masked.

There are various ways of expressing the amount or the concentration of analyte in the sample.
The information required and the method employed will dictate the methods of expressing
analytical results. Results will nearly always be reported as concentration on either weight or
volume basis, that is, the quantity of analyte per unit weight or per unit volume.

NOTE
 The units used are;

Weight: gram (g), Kilogram (kg= 103g), milligram (mg= 10-3g), microgram ( g-=10-6g),
nanogram (ng=10-9g)
Volume: litre(l), millitre (ml=10-3l), microliter ( l=10-6=-10-3ml), nanoliter (nl=10-9=10-6gml)
Concentration: Concentration include parts per thousand (ppt), parts per million (ppm), parts
per billion (ppb), molarity (M), Normality (N).

 You should be able to make the relevant conversions with these units

2.6 Calculation and interpretation of analytical data

Since measurement is a central phenomenon in analytical chemistry, evaluation of the analytical

procedure is one area that is important. Analytical procedures are evaluated through the
calculation and interpretation of analytical data. This is important because it is a measure of
performance and confidence of the analytical procedure. The performance of the analytical
procedure deals with its competence such as accuracy, while confidence deals with reliability.
This is measured by carrying out statistical analysis that recognizes change in composition or
performance due to changes or differences in set of readings.

Note

 Good laboratory records are essential in analytical chemistry. This ensures that the correct
date is used for calculation. This is important when communicating the validity of the results.

ACTIVITY

 Define the term chemical sample and chemical analysis.

 How should the laboratory records book be maintained?

SUMMARY
 In this lesson we have learnt the steps involved in carrying out a chemical analysis. This
includes obtaining a representative sample, sample preparation, measurement and lastly
calculating and interpreting data.